Patent application title: NOVEL METHODS FOR INDUCING AN IMMUNE RESPONSE
Inventors:
Marie-Ange DemoitiÉ (Rixensart, BE)
Marie-Noëlle Donner (Rixensart, BE)
Nadia Ouaked (Rixensart, BE)
Assignees:
GlaxoSmithKline Biologicals, s.a.
IPC8 Class: AA61K3904FI
USPC Class:
1 1
Class name:
Publication date: 2021-12-16
Patent application number: 20210386846
Abstract:
The present invention relates to methods for inducing an immune response,
in particular methods for inducing an immune response against
mycobacterial infections or disease comprising (i) at least one
administration of a polypeptide Rv1196 related antigen and at least one
administration of an adenovirus encoding a Rv1196 related antigen or (ii)
at least one administration of a polypeptide Rv0125 related antigen and
at least one administration of an adenovirus encoding a Rv0125 related
antigen. Associated compositions, adenoviral constructs and
polynucleotide sequences are also provided.Claims:
1-90. (canceled)
91. A method for inducing an immune response in a subject comprising: (i) administration of a polypeptide Rv0125 related antigen to the subject, followed by administration of a non-human simian adenovirus encoding a Rv0125 related antigen; or (ii) administration of a non-human simian adenovirus encoding a Rv0125 related antigen, followed by administration of a polypeptide Rv0125 related antigen to the subject.
92. The method of claim 91 wherein the polypeptide Rv0125 related antigen comprises a sequence having at least 90% identity to SEQ ID No: 3.
93. The method of claim 91 wherein the polypeptide Rv0125 related antigen comprises a fragment of SEQ ID No: 3 which is at least 150 amino acids in length.
94. The method of claim 91 wherein the encoded Rv0125 related antigen comprises a sequence having at least 90% identity to SEQ ID No: 3.
95. The method of claim 91 wherein the encoded Rv0125 related antigen comprises a fragment of SEQ ID No: 3 which is at least 150 amino acids in length.
96. The method of claim 91, wherein the polypeptide Rv0125 related antigen is provided in a composition further comprising an adjuvant.
97. The method of claim 96, wherein the adjuvant comprises a Toll Like Receptor (TLR) agonist and/or an immunologically active saponin.
98. The method of claim 96 wherein the adjuvant comprises 3-O-deacylated monophosphoryl lipid A (3D-MPL).
99. The method of claim 96 wherein the adjuvant comprises QS21.
100. The method of claim 91 wherein the non-human simian adenovirus is a chimpanzee adenovirus.
101. The method of claim 100 wherein the chimpanzee adenovirus is a ChAd3.
102. The method of claim 100 wherein the chimpanzee adenovirus is a ChAd63.
103. The method of claim 91, wherein the Rv0125 related antigen comprises a sequence having at least 90% identity to SEQ ID NO: 6.
104. The method of claim 91, wherein a composition comprising the polypeptide Rv0125 related antigen is substantially free of a non-human simian adenovirus encoding the Rv0125 related antigen.
105. The method of claim 91, wherein a composition comprising the polypeptide Rv0125 related antigen does not include a non-human simian adenovirus encoding the Rv0125 related antigen.
Description:
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This is the U.S. National Stage application submitted under 35 U.S.C. .sctn. 371 for International Application No. PCT/EP2016/067622, filed Jul. 25, 2016, which claims priority to Application No. GB 1513176.6, filed Jul. 27, 2015 all of which are incorporated herein by reference in their entireties.
TECHNICAL FIELD
[0002] The present invention relates to methods for inducing an immune response, in particular methods for inducing an immune response against mycobacterial infections or disease comprising (i) at least one administration of a polypeptide Rv1196 related antigen and at least one administration of an adenovirus encoding a Rv1196 related antigen or (ii) at least one administration of a polypeptide Rv0125 related antigen and at least one administration of an adenovirus encoding a Rv0125 related antigen. Associated compositions, adenoviral constructs and polynucleotide sequences are also provided.
BACKGROUND OF THE INVENTION
[0003] Vaccination is one of the most effective methods for preventing infectious diseases. However, a single administration of an antigen is often not sufficient to confer optimal immunity and/or a long-lasting response. Approaches for establishing strong and lasting immunity to specific pathogens include addition of adjuvants to vaccines and/or repeated vaccination, i.e. boosting an immune response by administration of one or more further doses of antigen. Such further administrations may be performed with the same vaccine (homologous boosting) or with a different vaccine (heterologous boosting).
[0004] Tuberculosis (TB) is a chronic infectious disease caused by infection with Mycobacterium tuberculosis and other Mycobacterium species. It is a major disease in developing countries, as well as an increasing problem in developed areas of the world.
[0005] Mtb72f and M72 are fusion protein antigens derived from the Mycobacterium tuberculosis proteins Rv1196 and Rv0125. Mtb72f and M72 (described, for example, in international patent applications WO2006/117240, WO2012/080369 and WO2012/080370 which are incorporated herein by reference) or fragments or derivatives thereof are protein antigens of potential benefit for the treatment or prevention of tuberculosis.
[0006] Preclinical and clinical investigations have led to M72 being administered in humans in conjunction with the immunostimulants 3-O-deacylated monophosphoryl lipid A (3D-MPL) and QS21 in a liposomal formulation and in a 0.1 month schedule using 10 ug M72 polypeptide, 25 ug 3D-MPL and 25 ug QS21 (Leroux-Roels et al Vaccine 2013 31 2196-2206, Montoya et al J. Clin. Immunol. 2013 33 (8): 1360-1375; Thacher E G et al AIDS 2014 28 (12):1769-1781; Idoko O T et al Tuberculosis (Edinb) 2014 94 (6):564-578; Penn-Nicholson A, et al Vaccine 2015 33 (32):4025-4034 doi:10.1016/j.vaccine.2015.05.088). A candidate vaccine utilising the M72 antigen is currently in a Phase IIB trial (ClinicalTrials.gov Identifier: NCT01755598) to evaluate the protective efficacy of two doses of adjuvanted protein against pulmonary TB, as compared to placebo, in adults aged 18-50 living in TB endemic countries.
[0007] WO2008107370A1 (incorporated herein by reference) describes the concomitant administration of a polypeptide antigen and an adenovirus encoding a polypeptide antigen.
[0008] WO2010023260 (incorporated herein by reference) describes the concomitant administration of a polypeptide antigen and viral vector encoding a polypeptide antigen.
[0009] There remains a need for novel methods of immunising against diseases, including tuberculosis, which are highly efficacious, safe, convenient, cost-effective, long-lasting and induce a broad spectrum of immune responses.
SUMMARY OF THE INVENTION
[0010] It has now surprisingly been found that, priming with a polypeptide Rv1196/Rv0125 related antigen and boosting with a non-human simian adenovirus encoding a Rv1196/Rv0125 related antigen, or priming with a non-human simian adenovirus encoding a Rv1196/Rv0125 related antigen with and boosting with a polypeptide Rv1196/Rv0125 related antigen can provide immune responses which are substantially improved relative to other potential approaches.
[0011] Accordingly, in a first aspect of the invention, there is provided a method for inducing an immune response in a subject comprising administration of a polypeptide Rv1196 related antigen to the subject, followed by administration of a non-human simian adenovirus encoding a Rv1196 related antigen.
[0012] In a second aspect of the invention, there is provided a method for inducing an immune response in a subject comprising administration of a non-human simian adenovirus encoding a Rv1196 related antigen, followed by administration of a polypeptide Rv1196 related antigen to the subject.
[0013] Suitably the polypeptide Rv1196 related antigen is provided in a composition which also comprises an adjuvant. Optionally, the adjuvant comprises a TLR agonist and/or an immunologically active saponin. The TLR agonist is suitably a TLR4 agonist.
[0014] Suitably the polypeptide Rv1196 related antigen is provided in a composition which does not comprise a non-human simian adenovirus encoding a Rv1196 related antigen.
[0015] Suitably the non-human simian adenovirus encoding a Rv1196 related antigen is provided in a composition which does not comprise a polypeptide Rv1196 related antigen.
[0016] In a third aspect of the invention, there is provided a method for inducing an immune response in a subject comprising administration of a polypeptide Rv0125 related antigen to the subject, followed by administration of a non-human simian adenovirus encoding a Rv0125 related antigen.
[0017] In a fourth aspect of the invention, there is provided a method for inducing an immune response in a subject comprising administration of a non-human simian adenovirus encoding a Rv0125 related antigen, followed by administration of a polypeptide Rv0125 related antigen to the subject.
[0018] Suitably the polypeptide Rv0125 related antigen is provided in a composition which also comprises an adjuvant. Optionally, the adjuvant comprises a TLR agonist and/or an immunologically active saponin. The TLR agonist is suitably a TLR4 agonist.
[0019] Suitably the polypeptide Rv0125 related antigen is provided in a composition which does not comprise a non-human simian adenovirus encoding a Rv0125 related antigen.
[0020] Suitably the non-human simian adenovirus encoding a Rv0125 related antigen is provided in a composition which does not comprise a polypeptide Rv0125 related antigen.
BRIEF DESCRIPTION OF THE FIGURES
[0021] FIG. 1: Median percentage of M72-specific CD4 T cell response from CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 7, 14 and 21 days post immunisation using ChAd3 in a range of doses.
[0022] FIG. 2: Median percentage of M72-specific CD4 T cell response from CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 7, 14 and 21 days post immunisation using ChAd63 in a range of doses.
[0023] FIG. 3: Median percentage of M72-specific CD8 T cell response from CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 7, 14 and 21 days post immunisation using ChAd3 in a range of doses.
[0024] FIG. 4: Median percentage of M72-specific CD8 T cell response from CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 7, 14 and 21 days post immunisation using ChAd63 in a range of doses.
[0025] FIG. 5: Percentage of M72-specific CD4 T cell response from whole blood of CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 14 PI and 14PII using heterologous (ChAd/Prot and Prot/ChAd) prime-boost vaccination strategies.
[0026] FIG. 6: Percentage of M72-specific CD4 T cell response from whole blood of CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 14 PI and 14PII using homologous (ChAd/ChAd) and in combination with M72/AS01E (admixture or co-administration) prime-boost vaccination strategies.
[0027] FIG. 7: Percentage of M72-specific CD8 T cell response from whole blood of CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 14 PI and 14PII using heterologous (ChAd/Prot and Prot/ChAd) prime-boost vaccination strategies.
[0028] FIG. 8: Percentage of M72-specific CD8 T cell response from whole blood of CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 14 PI and 14PII using homologous (ChAd/ChAd) and in combination with M72/AS01E (admixture or co-administration) prime-boost vaccination strategies.
[0029] FIG. 9: Percentage of M72-specific CD4 T cell response from lung tissue of CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 14PII.
[0030] FIG. 10: Percentage of M72-specific CD8 T cell response from lung tissue of CB6F1 mice expressing IFN-gamma and/or IL-2 and/or TNF-alpha cytokines at 14PII.
[0031] FIG. 11: Cytokine profile of the M72-specific CD4 T cells response in WBLO at 14PI in immunised CB6F1 mice.
[0032] FIG. 12: Table showing the data values of the cytokine profile of the M72-specific CD4 T cells response in WBLO at 14PI in immunised CB6F1 mice.
[0033] FIG. 13: Cytokine profile of the M72-specific CD4 T cells response in WBLO at 14PII in immunised CB6F1 mice.
[0034] FIG. 14: Table showing the data values of the cytokine profile of the M72-specific CD4 T cells response in WBLO at 14PII in immunised CB6F1 mice.
[0035] FIG. 15: Cytokine profile of the M72-specific CD4 T cells response in lung at 14PII in immunised CB6F1 mice.
[0036] FIG. 16: Table showing the data values of the cytokine profile of the M72-specific CD4 T cells response in lung at 14PII in immunised CB6F1 mice.
[0037] FIG. 17: Cytokine profile of the M72-specific CD8 T cells response in WBLO at 14PI in immunised CB6F1 mice.
[0038] FIG. 18: Table showing the data values of the cytokine profile of the M72-specific CD8 T cells response in WBLO at 14PI in immunised CB6F1 mice.
[0039] FIG. 19: Cytokine profile of the M72-specific CD8 T cells response in WBLO at 14PII in immunised CB6F1 mice.
[0040] FIG. 20: Table showing the data values of the cytokine profile of the M72-specific CD8 T cells response in WBLO at 14PII in immunised CB6F1 mice.
[0041] FIG. 21: Cytokine profile of the M72-specific CD8 T cells response in lung at 14PII in immunised CB6F1 mice.
[0042] FIG. 22: Table showing the data values of the cytokine profile of the M72-specific CD8 T cells response in lung at 14PII in immunised CB6F1 mice.
[0043] FIG. 23: Anti-M72 Ig tot serology at 13dPII.
[0044] FIG. 24: Diagrammatic representation of M72-ChAd3 construct arrangement
[0045] FIG. 25: Diagrammatic representation of M72-ChAd63 construct arrangement
BRIEF DESCRIPTION OF THE SEQUENCE IDENTIFIERS
[0046] SEQ ID No: 1 Mycobacterium tuberculosis H37Rv Rv1196 polypeptide sequence
[0047] SEQ ID No: 2 Mycobacterium tuberculosis F11 Rv1196 polypeptide sequence
[0048] SEQ ID No: 3 Mycobacterium tuberculosis H37Rv Rv0125 polypeptide sequence (mature sequence)
[0049] SEQ ID No: 4 M72 2-his polypeptide sequence
[0050] SEQ ID No: 5 M72 2-his polynucleotide
[0051] SEQ ID No: 6 M72 No his polypeptide sequence
[0052] SEQ ID No: 7 M72 No his polynucleotide
[0053] SEQ ID No: 8 M72 No his human optimised polynucleotide
[0054] SEQ ID No: 9 ChAd3 polynucleotide
[0055] SEQ ID No: 10 ChAd3 penton polypeptide sequence
[0056] SEQ ID No: 11 ChAd3 hexon polypeptide sequence
[0057] SEQ ID No: 12 ChAd3 fibre polypeptide sequence
[0058] SEQ ID No: 13 M72-ChAd3 construct DNA
[0059] SEQ ID No: 14 ChAd63 polynucleotide
[0060] SEQ ID No: 15 ChAd63 penton polypeptide sequence
[0061] SEQ ID No: 16 ChAd63 hexon polypeptide sequence
[0062] SEQ ID No: 17 ChAd63 fibre polypeptide sequence
[0063] SEQ ID No: 18 M72-ChAd63 construct DNA
[0064] SEQ ID No: 19 ChAd155 polynucleotide
[0065] SEQ ID No: 20 ChAd155 penton polypeptide sequence
[0066] SEQ ID No: 21 ChAd155 hexon polypeptide sequence
[0067] SEQ ID No: 22 ChAd155 fibre polypeptide sequence
DETAILED DESCRIPTION
[0068] Tuberculosis (TB) is a chronic infectious disease caused by infection with Mycobacterium tuberculosis and other Mycobacterium species. It is a major disease in developing countries, as well as an increasing problem in developed areas of the world. About one third of the world's population are believed to be latently infected with TB bacilli, with about 9 million new cases of active TB and 1.5 million deaths each year. Around 10% of those infected with TB bacilli will develop active TB, each person with active TB infecting an average of 10 to 15 others per year. (World Health Organisation Tuberculosis Facts 2014)
[0069] Mycobacterium tuberculosis infects individuals through the respiratory route. Alveolar macrophages engulf the bacterium, but it is able to survive and proliferate by inhibiting phagosome fusion with acidic lysosomes. A complex immune response involving CD4+ and CD8+T cells ensues, ultimately resulting in the formation of a granuloma. Central to the success of Mycobacterium tuberculosis as a pathogen is the fact that the isolated, but not eradicated, bacterium may persist for long periods, leaving an individual vulnerable to the later development of active TB.
[0070] Fewer than 5% of infected individuals develop active TB in the first years after infection. The granuloma can persist for decades and is believed to contain live Mycobacterium tuberculosis in a state of dormancy, deprived of oxygen and nutrients. However, it has been suggested that the majority of the bacteria in the dormancy state are located in non-macrophage cell types spread throughout the body (Locht et al, Expert Opin. Biol. Ther. 2007 7 (11):1665-1677). The development of active TB occurs when the balance between the host's natural immunity and the pathogen changes, for example as a result of an immunosuppressive event (Anderson P Trends in Microbiology 2007 15 (1):7-13; Ehlers S Infection 2009 37 (2):87-95).
[0071] A dynamic hypothesis describing the balance between latent TB and active TB has also been proposed (Cardona P-J Inflammation & Allergy--Drug Targets 2006 6:27-39; Cardona P-J Infection 2009 37 (2):80-86).
[0072] Although an infection may be asymptomatic for a considerable period of time, the active disease is most commonly manifested as an acute inflammation of the lungs, resulting in tiredness, weight loss, fever and a persistent cough. If untreated, serious complications and death typically result.
[0073] Tuberculosis can generally be controlled using extended antibiotic therapy, although such treatment is not sufficient to prevent the spread of the disease. Actively infected individuals may be largely asymptomatic, but contagious, for some time. In addition, although compliance with the treatment regimen (which typically lasts 6 months or more) is critical, patient behaviour is difficult to monitor. Some patients do not complete the course of treatment, which can lead to ineffective treatment and the development of drug resistance.
[0074] Multidrug-resistant TB (MDR-TB) is a form which fails to respond to first line medications. An estimated 480,000 people developed MDR-TB in 2013. MDR-TB is treatable by using second-line drugs. However, second-line treatment options are limited and recommended medicines are not always available. The extensive chemotherapy required (up to two years of treatment) is costly and can produce severe adverse drug reactions in patients.
[0075] Extensively drug-resistant TB (XDR-TB) occurs when resistance to second line medications develops on top of resistance to first line medications. It is estimated that about 9.0% of MDR-TB cases had XDR-TB (World Health Organisation Tuberculosis Facts 2014).
[0076] Even if a full course of antibiotic treatment is completed, infection with M. tuberculosis may not be eradicated from the infected individual and may remain as a latent infection that can be reactivated. Consequently, accurate and early diagnosis of the disease are of utmost importance.
[0077] Currently, vaccination with attenuated live bacteria is the most widely used method for inducing protective immunity. The most common Mycobacterium employed for this purpose is Bacillus Calmette-Guerin (BCG), an avirulent strain of M. bovis which was first developed over 60 years ago. It is administrated at birth in TB endemic regions. However, the safety and efficacy of BCG is a source of controversy--while protecting against severe disease manifestation in children, the efficacy of BCG against disease in adults is variable. Additionally, some countries, such as the United States, do not vaccinate the general public with this agent.
[0078] Several of the proteins which are strongly expressed during the early stages of Mycobacterium infection have been shown to provide protective efficacy in animal vaccination models. However, vaccination with antigens which are highly expressed during the early stages of infection may not provide an optimal immune response for dealing with later stages of infection. Adequate control during latent infection may require T cells which are specific for the particular antigens which are expressed at that time. Post-exposure vaccines which directly target the dormant persistent bacteria may aid in protecting against TB reactivation, thereby enhancing TB control, or even enabling clearance of the infection. A vaccine targeting latent TB could therefore significantly and economically reduce global TB infection rates.
[0079] Subunit vaccines based on late stage antigens could also be utilised in combination with early stage antigens to provide a multiphase vaccine. Alternatively, early and/or late stage antigens could be used to complement and improve BCG vaccination (either by boosting the BCG response or through the development of advanced recombinant BCG strains).
[0080] Mtb72f and M72 are fusion protein antigens of potential benefit for the treatment or prevention of tuberculosis. Mtb72f and M72 are derived from the Mycobacterium tuberculosis proteins Rv1196 and Rv0125, both genes are present in both virulent and avirulent strains of the Mycobacterium tuberculosis complex, and in BCG.
[0081] Rv1196 (described, for example, by the name Mtb39a in Dillon et al Infection and Immunity 1999 67 (6): 2941-2950) is highly conserved, with 100% sequence identity across H37Rv, C, Haarlem, CDC1551, 94-M4241A, 98-R6041NH-RIF-EM, KZN605, KZN1435, KZN4207, KZNR506 strains, the F11 strain having a single point mutation Q30K (most other clinical isolates have in excess of 90% identity to H37Rv). An adenovirus encoding an Rv1196 related antigen is described in Lewinsohn et al Am J Respir Crit Care Med 2002 116:843-848.
[0082] Rv0125 (described, for example, by the name Mtb32a in Skeiky et al Infection and Immunity 1999 67 (8): 3998-4007) is also highly conserved, with 100% sequence identity across many strains. An adenovirus (human Ad5) encoding an Rv0125 related antigen is described in Zhang et al Human Vaccines & Therapeutics 2015 11 (7):1803-1813 doi: 10.1080/21645515.2015.1042193. Full length Rv0125 includes an N-terminal signal sequence which is cleaved to provide the mature protein.
[0083] Mtb72f has been shown to provide protection in a number of animal models (see, for example: Brandt et al Infect. Immun. 2004 72 (11):6622-6632; Skeiky et al J. Immunol. 2004 172:7618-7628; Tsenova et al Infect. Immun. 2006 74 (4):2392-2401). Mtb72f has also been the subject of clinical investigations (Von Eschen et al 2009 Human Vaccines 5 (7):475-482). M72 is an improved antigen which incorporates a single serine to alanine mutation relative to Mtb72f, resulting in improved stability characteristics. M72 related antigens have also been shown to be of value in a latent TB model (international patent application WO2006/117240, incorporated herein by reference). Previous pre-clinical and clinical investigations have led to M72 being administered in humans in conjunction with the immunostimulants 3-O-deacylated monophosphoryl lipid A (3D-MPL) and QS21 in a liposomal formulation and in a 0.1 month schedule using 10 ug M72 polypeptide, 25 ug 3D-MPL and 25 ug QS21 (see, for example, Leroux-Roels et al Vaccine 2013 31 2196-2206, Montoya et al J. Clin. Immunol. 2013 33 (8): 1360-1375; Thacher E G et al AIDS 2014 28 (12):1769-1781; Idoko O T et al Tuberculosis (Edinb) 2014 94 (6):564-578; Penn-Nicholson A, et al Vaccine 2015 33 (32):4025-4034 doi:10.1016/j.vaccine.2015.05.088).
[0084] A candidate vaccine utilising the antigen M72 is currently in a Phase IIB trial (ClinicalTrials.gov Identifier: NCT01755598) to evaluate the protective efficacy of two doses doses of adjuvanted protein against pulmonary TB, as compared to placebo, in adults aged 18-50 living in TB endemic countries. Nevertheless, a need for improved vaccination approaches remains.
[0085] In a first aspect of the invention, there is provided a method for inducing an immune response in a subject comprising administration of a polypeptide Rv1196 related antigen to the subject, followed by administration of a non-human simian adenovirus encoding a Rv1196 related antigen.
[0086] In a second aspect of the invention, there is provided a method for inducing an immune response in a subject comprising administration of a non-human simian adenovirus encoding a Rv1196 related antigen to the subject, followed by administration of a polypeptide Rv1196 related antigen.
[0087] In a third aspect of the invention, there is provided a method for inducing an immune response in a subject comprising administration of a polypeptide Rv0125 related antigen to the subject, followed by administration of an immunogenic composition comprising a non-human simian adenovirus encoding a Rv0125 related antigen.
[0088] In a fourth aspect of the invention, there is provided a method for inducing an immune response in a subject comprising administration of a non-human simian adenovirus encoding a Rv0125 related antigen, followed by administration of a polypeptide Rv0125 related antigen to the subject.
[0089] As used herein, administration of a first composition "followed by" administration of a second composition indicates that a time interval has elapsed between administration of the first composition and administration of the second composition. Suitably the time interval between administrations is one week to two years, in particular two weeks to eighteen months, typically three weeks to fifteen months, such as three weeks to six months, for example three weeks to two months, especially three weeks to six weeks such as around four weeks.
[0090] Also provided is a polypeptide Rv1196 related antigen, for use in inducing an immune response in a subject wherein the polypeptide Rv1196 related antigen is administered to the subject, followed by the administration of a non-human simian adenovirus encoding a Rv1196 related antigen.
[0091] Similarly, there is provided a non-human simian adenovirus encoding a Rv1196 related antigen, for use in inducing an immune response in a subject wherein a polypeptide Rv1196 related antigen is administered to the subject, followed by the administration of the non-human simian adenovirus encoding a Rv1196 related antigen.
[0092] Further, there is provided the use of a polypeptide Rv1196 related antigen, in the manufacture of a medicament for inducing an immune response in a subject wherein the polypeptide Rv1196 related antigen is administered to the subject, followed by the administration of a non-human simian adenovirus encoding a Rv1196 related antigen.
[0093] Additionally, there is provided the use of a non-human simian adenovirus encoding a Rv1196 related antigen, in the manufacture of a medicament for inducing an immune response in a subject wherein a polypeptide Rv1196 related antigen is administered to the subject, followed by the administration of the non-human simian adenovirus encoding a Rv1196 related antigen.
[0094] Also provided is a polypeptide Rv1196 related antigen, for use in inducing an immune response in a subject wherein a non-human simian adenovirus encoding a Rv1196 related antigen is administered to the subject, followed by the administration of the polypeptide Rv1196 related antigen.
[0095] Similarly, there is provided a non-human simian adenovirus encoding a Rv1196 related antigen, for use in inducing an immune response in a subject wherein the non-human simian adenovirus encoding a Rv1196 related antigen is administered to the subject, followed by the administration of a polypeptide Rv1196 related antigen.
[0096] Further, there is provided the use of a polypeptide Rv1196 related antigen, in the manufacture of a medicament for inducing an immune response in a subject wherein a non-human simian adenovirus encoding a Rv1196 related antigen is administered to the subject, followed by the administration of the polypeptide Rv1196 related antigen.
[0097] Also provided is the use of a non-human simian adenovirus encoding a Rv1196 related antigen, in the manufacture of a medicament for inducing an immune response in a subject wherein the non-human simian adenovirus encoding a Rv1196 related antigen is administered to the subject, followed by the administration of a polypeptide Rv1196 related antigen.
[0098] Also provided is a polypeptide Rv0125 related antigen, for use in inducing an immune response in a subject wherein the polypeptide Rv0125 related antigen is administered to the subject, followed by the administration of a non-human simian adenovirus encoding a Rv0125 related antigen.
[0099] Similarly, there is provided a non-human simian adenovirus encoding a Rv0125 related antigen, for use in inducing an immune response in a subject wherein a polypeptide Rv0125 related antigen is administered to the subject, followed by the administration of the non-human simian adenovirus encoding a Rv0125 related antigen.
[0100] Further, there is provided the use of a polypeptide Rv0125 related antigen, in the manufacture of a medicament for inducing an immune response in a subject wherein the polypeptide Rv0125 related antigen is administered to the subject, followed by the administration of a non-human simian adenovirus encoding a Rv0125 related antigen.
[0101] Additionally, there is provided the use of a non-human simian adenovirus encoding a Rv0125 related antigen, in the manufacture of a medicament for inducing an immune response in a subject wherein a polypeptide Rv0125 related antigen is administered to the subject, followed by the administration of the non-human simian adenovirus encoding a Rv0125 related antigen.
[0102] Also provided is a polypeptide Rv0125 related antigen, for use in inducing an immune response in a subject wherein a non-human simian adenovirus encoding a Rv0125 related antigen is administered to the subject, followed by the administration of the polypeptide Rv0125 related antigen.
[0103] Similarly, there is provided a non-human simian adenovirus encoding a Rv0125 related antigen, for use in inducing an immune response in a subject wherein the non-human simian adenovirus encoding a Rv0125 related antigen is administered to the subject, followed by the administration of a polypeptide Rv0125 related antigen.
[0104] Further, there is provided the use of a polypeptide Rv0125 related antigen, in the manufacture of a medicament for inducing an immune response in a subject wherein a non-human simian adenovirus encoding a Rv0125 related antigen is administered to the subject, followed by the administration of the polypeptide Rv0125 related antigen.
[0105] Also provided is the use of a non-human simian adenovirus encoding a Rv0125 related antigen, in the manufacture of a medicament for inducing an immune response in a subject wherein the non-human simian adenovirus encoding a Rv0125 related antigen is administered to the subject, followed by the administration of a polypeptide Rv0125 related antigen.
[0106] Suitably the polypeptide Rv1196 related antigen is provided in a composition which also comprises an adjuvant. Optionally, the adjuvant comprises a TLR agonist and/or an immunologically active saponin. The TLR agonist is suitably a TLR4 agonist.
[0107] Suitably the polypeptide Rv0125 related antigen is provided in a composition which also comprises an adjuvant. Optionally, the adjuvant comprises a TLR agonist and/or an immunologically active saponin. The TLR agonist is suitably a TLR4 agonist.
[0108] Suitably the polypeptide Rv1196 related antigen is provided in a composition which is substantially free of a non-human simian adenovirus encoding a Rv1196 related antigen, such as it does not comprise a non-human simian adenovirus encoding a Rv1196 related antigen. For example, it is substantially free of or does not comprise a non-human simian adenovirus encoding a mycobacterial antigen (such as it is substantially free of or does not comprise any non-human simian adenovirus) in particular it is substantially free of or does not comprise any adenovirus encoding a mycobacterial antigen (such as it is substantially free of or does not comprise any adenovirus). In some embodiments the polypeptide Rv1196 related antigen is provided in a composition which is substantially free of or does not comprise any adenovirus encoding a mycobacterial antigen.
[0109] Furthermore, the polypeptide Rv1196 related antigen is suitably not administered within a period of one day (such as two, three or six days) of a non-human simian adenovirus encoding a Rv1196 related antigen, such a non-human simian adenovirus encoding a Rv1196 related antigen. For example, a non-human simian adenovirus encoding a mycobacterial antigen (such as any non-human simian adenovirus) in particular any adenovirus encoding a mycobacterial antigen (such as any adenovirus). The polypeptide Rv1196 related antigen is suitably not administered within a period of one day (such as two, three or six days) of any adenovirus encoding a mycobacterial antigen
[0110] Suitably the non-human simian adenovirus encoding a Rv1196 related antigen is provided in a composition which is substantially free of or does not comprise a polypeptide Rv1196 related antigen. For example, it is substantially free of or does not comprise a polypeptide mycobacterial antigen (such as is substantially free of or it does not comprise any other antigens). In some embodiments the non-human simian adenovirus encoding a Rv1196 related antigen is provided in a composition which is substantially free of or does not comprise a polypeptide mycobacterial antigen.
[0111] Furthermore, the non-human simian adenovirus encoding a Rv1196 related antigen is suitably not administered within a period of one day (such as two, three or six days) of a polypeptide Rv1196 related antigen. For example, a polypeptide mycobacterial antigen (such as any other antigens). The non-human simian adenovirus encoding a Rv1196 related antigen is suitably not administered within a period of one day (such as two, three or six days) of a polypeptide mycobacterial antigen.
[0112] Suitably the polypeptide Rv0125 related antigen is provided in a composition which is substantially free of a non-human simian adenovirus encoding a Rv0125 related antigen, such as it does not comprise a non-human simian adenovirus encoding a Rv0125 related antigen. For example, it is substantially free of or does not comprise a non-human simian adenovirus encoding a mycobacterial antigen (such as it is substantially free of or does not comprise any non-human simian adenovirus) in particular it is substantially free of or does not comprise any adenovirus encoding a mycobacterial antigen (such as it is substantially free of or does not comprise any adenovirus). In some embodiments the polypeptide Rv0125 related antigen is provided in a composition which is substantially free of or does not comprise any adenovirus encoding a mycobacterial antigen.
[0113] Furthermore, the polypeptide Rv1196 related antigen is suitably not administered within a period of one day (such as two, three or six days) of a non-human simian adenovirus encoding a Rv0125 related antigen, such a non-human simian adenovirus encoding a Rv1196 related antigen. For example, a non-human simian adenovirus encoding a mycobacterial antigen (such as any non-human simian adenovirus) in particular any adenovirus encoding a mycobacterial antigen (such as any adenovirus). The polypeptide Rv0125 related antigen is suitably not administered within a period of one day (such as two, three or six days) of any adenovirus encoding a mycobacterial antigen
[0114] Suitably the non-human simian adenovirus encoding a Rv0125 related antigen is provided in a composition which is substantially free of or does not comprise a polypeptide Rv0125 related antigen. For example, it is substantially free of or does not comprise a polypeptide mycobacterial antigen (such as is substantially free of or it does not comprise any other antigens). In some embodiments the non-human simian adenovirus encoding a Rv0125 related antigen is provided in a composition which is substantially free of or does not comprise a polypeptide mycobacterial antigen.
[0115] Furthermore, the non-human simian adenovirus encoding a Rv0125 related antigen is suitably not administered within a period of one day (such as two, three or six days) of a polypeptide Rv1196 related antigen. For example, a polypeptide mycobacterial antigen (such as any other antigens). The non-human simian adenovirus encoding a Rv0125 related antigen is suitably not administered within a period of one day (such as two, three or six days) of a polypeptide mycobacterial antigen.
[0116] Suitably, the subject is a mammal, such as a bovine or human. In particular, the subject is a human.
[0117] By substantially free of, in the context of adenovirus, typically means comprising less than 10.sup.4, such as less than 10.sup.3, in particular less than 10.sup.2 or less than 10.sup.1 viral particles of the relevant type per dose. Suitable methods for determining the number of viral particles include Quantitative PCR Analysis, analytical HLPC or spectrophotometric methods based on A.sub.260 nm.
[0118] By substantially free of, in the context of polypeptide antigen, typically means comprising less than 1 ug, such as less than 0.1 ug, in particular less than 0.01 ug of the relevant antigen or antigens per dose. Suitable methods for determining the amount of peptide antigen are known to the skilled person and depending on the composition, may combine purification methods to assist in facilitating an appropriate quantification method (such as analytical HLPC or spectrophotometric methods).
[0119] Typically, the aim of the method of the invention is to induce a protective immune response, i.e. immunise or vaccinate the subject against a related pathogen. The invention may therefore be applied for the prophylaxis, treatment or amelioration of infection by mycobacteria, such as infection by Mycobacterium bovis or Mycobacterium tuberculosis, in particular Mycobacterium tuberculosis.
[0120] The invention may be provided for the purpose of:
[0121] prophylaxis of active tuberculosis due to infection (i.e. primary tuberculosis) or reactivation (i.e. secondary tuberculosis), such as by administering to a subject who is uninfected, or alternatively a subject who has a latent infection;
[0122] prophylaxis of latent tuberculosis, such as by administering to a subject who is uninfected;
[0123] treating latent tuberculosis;
[0124] preventing or delaying reactivation of tuberculosis, especially the delay of reactivation, for example by a period of months, years or indefinitely; or
[0125] treating active tuberculosis (such as to reduce the need for chemotherapeutic treatment: such as reduced term of chemotherapeutic treatment, complexity of drug regimen or dosage of chemotherapeutic treatment; alternatively, to reduce the risk of a later relapse following chemotherapeutic treatment).
[0126] The elicited immune response may be an antigen specific T cell response (which may be a systemic and/or a local response). Systemic responses may be detected, for example, from a sample of whole blood. Local responses (for example, the local response in the lung) may be detected from an appropriate sample of tissue (for example, lung tissue) or other locally focused samply method (e.g. bronchoalveolar lavage). The antigen specific T cell response may comprise a CD4+T cell response, such as a response involving CD4+T cells expressing a plurality of cytokines (e.g. IFNgamma, TNFalpha or IL2, especially IFNgamma, TNFalpha and IL2). Alternatively, or additionally, the antigen specific T cell response comprises a CD8+T cell response, such as a response involving CD8+T cells expressing a plurality of cytokines (e.g. IFNgamma, TNFalpha or IL2, especially IFNgamma, TNFalpha and IL2).
[0127] The term "active infection" refers to an infection, e.g. infection by M. tuberculosis, with manifested disease symptoms and/or lesions, suitably with manifested disease symptoms.
[0128] The terms "inactive infection", "dormant infection" or "latent infection" or "latent tuberculosis" refer to an infection, e.g. infection by M. tuberculosis, without manifested disease symptoms and/or lesions, suitably without manifested disease symptoms. A subject with latent infection will suitably be one which tests positive for infection, e.g. by Tuberculin skin test (TST) or Interferon-Gamma Release Assays (IGRAs), but which has not demonstrated the disease symptoms and/or lesions which are associated with an active infection.
[0129] The term "primary tuberculosis" refers to clinical illness, e.g., manifestation of disease symptoms, directly following infection, e.g. infection by M. tuberculosis. See, Harrison's Principles of Internal Medicine, Chapter 150, pp. 953-966 (16th ed., Braunwald, et al., eds., 2005).
[0130] The terms "secondary tuberculosis" or "postprimary tuberculosis" refer to the reactivation of a dormant, inactive or latent infection, e.g. infection by M. tuberculosis. See, Harrison's Principles of Internal Medicine, Chapter 150, pp. 953-966 (16th ed., Braunwald, et al., eds., 2005).
[0131] The term "tuberculosis reactivation" refers to the later manifestation of disease symptoms in an individual that tests positive for infection (e.g. by Tuberculin skin test (TST) or Interferon-Gamma Release Assays (IGRAs)) but does not have apparent disease symptoms. Suitably the individual will not have been re-exposed to infection. The positive diagnostic test indicates that the individual is infected, however, the individual may or may not have previously manifested active disease symptoms that had been treated sufficiently to bring the tuberculosis into an inactive or latent state.
[0132] Suitability the methods are applied to a subject who is uninfected or who has a latent infection by mycobacteria, such as infection by Mycobacterium tuberculosis. In one embodiment the methods are applied to a subject who does not have an infection by Mycobacterium tuberculosis (in the context of human subjects) or Mycobacterium bovis (in the context of bovine subjects). In another embodiment the methods are applied to a subject who has a latent infection by mycobacteria, such as Mycobacterium tuberculosis (in the context of human subjects) or Mycobacterium bovis (in the context of bovine subjects).
[0133] In some embodiments, the subject has previously been vaccinated with BCG. The approaches of the present invention may, for example, be utilised for a subject at least one year after BCG vaccination, for example at least two years after BCG vaccination such as at least at least five years after BCG vaccination.
[0134] In some embodiments, the subject has previously been infected with M. tuberculosis.
Antigens of Use in the Invention
[0135] T cell epitopes are short contiguous stretches of amino acids which are recognised by T cells (e.g. CD4+ or CD8+T cells). Identification of T cell epitopes may be achieved through epitope mapping experiments which are known to the person skilled in the art (see, for example, Paul, Fundamental Immunology, 3rd ed., 243-247 (1993); Bei.beta.barth et al Bioinformatics 2005 21 (Suppl. 1):i29-i37). In a diverse out-bred population, such as humans, different HLA types mean that particular epitopes may not be recognised by all members of the population. As a result of the crucial involvement of the T cell response in tuberculosis, to maximise the level of recognition and scale of immune response, an immunogenic derivative of a reference sequence is desirably one which contains the majority (or suitably all) T cell epitopes intact. Mortier et al BMC Immunology 2015 16:63 undertake sequence conservation analysis and in silico human leukocyte antigen-peptide binding predictions for Mtb72f and M72 tuberculosis candidate vaccine antigens.
[0136] The skilled person will recognise that individual substitutions, deletions or additions to a protein which alters, adds or deletes a single amino acid or a small percentage of amino acids is an "immunogenic derivative" where the alteration(s) results in the substitution of an amino acid with a functionally similar amino acid or the substitution/deletion/addition of residues which do not substantially impact the immunogenic function.
[0137] Conservative substitution tables providing functionally similar amino acids are well known in the art. In general, such conservative substitutions will fall within one of the amino-acid groupings specified below, though in some circumstances other substitutions may be possible without substantially affecting the immunogenic properties of the antigen. The following eight groups each contain amino acids that are typically conservative substitutions for one another:
[0138] 1) Alanine (A), Glycine (G);
[0139] 2) Aspartic acid (D), Glutamic acid (E);
[0140] 3) Asparagine (N), Glutamine (Q);
[0141] 4) Arginine (R), Lysine (K);
[0142] 5) Isoleucine (I), Leucine (L), Methionine (M), Valine (V);
[0143] 6) Phenylalanine (F), Tyrosine (Y), Tryptophan (W);
[0144] 7) Serine (S), Threonine (T); and
[0145] 8) Cysteine (C), Methionine (M)
[0146] (see, e.g., Creighton, Proteins 1984).
Suitably such substitutions do not occur in the region of an epitope, and do not therefore have a significant impact on the immunogenic properties of the antigen.
[0147] Immunogenic derivatives may also include those wherein additional amino acids are inserted compared to the reference sequence. Suitably such insertions do not occur in the region of an epitope, and do not therefore have a significant impact on the immunogenic properties of the antigen. One example of insertions includes a short stretch of histidine residues (e.g. 2-6 residues) to aid expression and/or purification of the antigen in question.
[0148] Immunogenic derivatives include those wherein amino acids have been deleted compared to the reference sequence. Suitably such deletions do not occur in the region of an epitope, and do not therefore have a significant impact on the immunogenic properties of the antigen.
[0149] The skilled person will recognise that a particular immunogenic derivative may comprise substitutions, deletions and additions (or any combination thereof).
[0150] The terms "identical" or percentage "identity," in the context of two or more polypeptide sequences, refer to two or more sequences or sub-sequences that are the same or have a specified percentage of amino acid residues that are the same (i.e., 70% identity, optionally 75%, 80%, 85%, 90%, 95%, 98% or 99% identity over a specified region), when compared and aligned for maximum correspondence over a comparison window, or designated region as measured using one of the following sequence comparison algorithms or by manual alignment and visual inspection. This definition also refers to the compliment of a test sequence. Optionally, the identity exists over a region that is at least 200 amino acids in length, such as at least 300 amino acids or at least 400 amino acids. Suitably, the comparison is performed over a window corresponding to the entire length of the reference sequence (as opposed to the derivative sequence).
[0151] For sequence comparison, one sequence acts as the reference sequence, to which the test sequences are compared. When using a sequence comparison algorithm, test and reference sequences are entered into a computer, subsequence coordinates are designated, if necessary, and sequence algorithm program parameters are designated. Default program parameters can be used, or alternative parameters can be designated. The sequence comparison algorithm then calculates the percentage sequence identities for the test sequences relative to the reference sequence, based on the program parameters.
[0152] A "comparison window", as used herein, refers to a segment in which a sequence may be compared to a reference sequence of the same number of contiguous positions after the two sequences are optimally aligned. Methods of alignment of sequences for comparison are well-known in the art. Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv. Appl. Math. 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J. Mol. Biol. 48:443 (1970), by the search for similarity method of Pearson & Lipman, Proc. Nat'l. Acad. Sci. USA 85:2444 (1988), by computerised implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, Wis.), or by manual alignment and visual inspection (see, e.g., Current Protocols in Molecular Biology (Ausubel et al., eds. 1995 supplement)).
[0153] One example of a useful algorithm is PILEUP. PILEUP creates a multiple sequence alignment from a group of related sequences using progressive, pairwise alignments to show relationship and percent sequence identity. It also plots a tree or dendogram showing the clustering relationships used to create the alignment. PILEUP uses a simplification of the progressive alignment method of Feng & Doolittle, J. Mol. Eva 35:351-360 (1987). The method used is similar to the method described by Higgins & Sharp, CABIOS 5:151-153 (1989). The program can align up to 300 sequences, each of a maximum length of 5,000 nucleotides or amino acids. The multiple alignment procedure begins with the pairwise alignment of the two most similar sequences, producing a cluster of two aligned sequences. This cluster is then aligned to the next most related sequence or cluster of aligned sequences. Two clusters of sequences are aligned by a simple extension of the pairwise alignment of two individual sequences. The final alignment is achieved by a series of progressive, pairwise alignments. The program is run by designating specific sequences and their amino acid coordinates for regions of sequence comparison and by designating the program parameters. Using PILEUP, a reference sequence is compared to other test sequences to determine the percent sequence identity relationship using the following parameters: default gap weight (3.00), default gap length weight (0.10), and weighted end gaps. PILEUP can be obtained from the GCG sequence analysis software package, e.g., version 7.0 (Devereaux et al., Nuc. Acids Res. 12:387-395 (1984)).
[0154] Another example of algorithm that is suitable for determining percent sequence identity and sequence similarity are the BLAST and BLAST 2.0 algorithms, which are described in Altschul et al., Nuc. Acids Res. 25:3389-3402 (1977) and Altschul et al., J. Mol. Biol. 215:403-410 (1990), respectively. Software for performing BLAST analyses is publicly available through the National Center for Biotechnology Information (website at www.ncbi.nlm.nih.gov/). This algorithm involves first identifying high scoring sequence pairs (HSPs) by identifying short words of length W in the query sequence, which either match or satisfy some positive-valued threshold score T when aligned with a word of the same length in a database sequence. T is referred to as the neighbourhood word score threshold (Altschul et al., supra). These initial neighbourhood word hits act as seeds for initiating searches to find longer HSPs containing them. The word hits are extended in both directions along each sequence for as far as the cumulative alignment score can be increased. Cumulative scores are calculated using, for nucleotide sequences, the parameters M (reward score for a pair of matching residues; always >0) and N (penalty score for mismatching residues; always <0). For amino acid sequences, a scoring matrix is used to calculate the cumulative score. Extension of the word hits in each direction are halted when: the cumulative alignment score falls off by the quantity X from its maximum achieved value; the cumulative score goes to zero or below, due to the accumulation of one or more negative-scoring residue alignments; or the end of either sequence is reached. The BLAST algorithm parameters W, T, and X determine the sensitivity and speed of the alignment. The BLASTN program (for nucleotide sequences) uses as defaults a wordlength (W) of 11, an expectation (E) or 10, M=5, N=-4 and a comparison of both strands. For amino acid sequences, the BLASTP program uses as defaults a wordlength of 3, and expectation (E) of 10, and the BLOSUM62 scoring matrix (see Henikoff & Henikoff, Proc. Natl. Acad. Sci. USA 89:10915 (1989)) alignments (B) of 50, expectation (E) of 10, M=5, N=-4, and a comparison of both strands.
[0155] The BLAST algorithm also performs a statistical analysis of the similarity between two sequences (see, e.g., Karlin & Altschul, Proc. Nat'l. Acad. Sci. USA 90:5873-5787 (1993)). One measure of similarity provided by the BLAST algorithm is the smallest sum probability (P(N)), which provides an indication of the probability by which a match between two nucleotide or amino acid sequences would occur by chance. For example, a nucleic acid is considered similar to a reference sequence if the smallest sum probability in a comparison of the test nucleic acid to the reference nucleic acid is less than about 0.2, more preferably less than about 0.01, and most preferably less than about 0.001.
[0156] In any event, immunogenic derivatives of a polypeptide sequence will usually have essentially the same activity as the reference sequence. By essentially the same activity is meant at least 50%, suitably at least 75% and especially at least 90% activity of the reference sequence in an in vitro restimulation assay of PBMC, whole blood, lung tissue or bronchoalveolar lavage with specific antigens (e.g. restimulation for a period of between several hours to up to two weeks, such as up to one day, 1 day to 1 week or 1 to 2 weeks) that measures the activation of the cells via lymphoproliferation, production of cytokines in the supernatant of culture (measured by ELISA, CBA etc) or characterisation of T and B cell responses by intra and extracellular staining (e.g. using antibodies specific to immune markers, such as CD3, CD4, CD8, IL2, TNF-alpha, IFN-gamma, IL-17, CD40L, CD69 etc) followed by analysis with a flow cytometer. Suitably, by essentially the same activity is meant at least 50%, suitably at least 75% and especially at least 90% activity of the reference sequence in a T cell proliferation and/or IFN-gamma production assay.
[0157] In one embodiment the polypeptide antigen and the encoded antigen are Rv1196 related antigens. The term `Rv1196 related antigen` refers to the Rv1196 protein provided in SEQ ID No: 1 or an immunogenic derivative thereof. As used herein the term "derivative" refers to an antigen that is modified relative to the reference sequence. Immunogenic derivatives are sufficiently similar to the reference sequence to substantially retain the immunogenic properties of the reference sequence and remain capable of allowing an immune response to be raised against the reference sequence. An immunogenic derivative may, for example, comprise a modified version of the reference sequence or alternatively may consist of a modified version of the reference sequence.
[0158] The Rv1196 related antigen may for example contain 2500 amino acid residues or fewer, such 1500 amino acid residues or fewer, in particular 1200 amino acid residues or fewer, especially 1000 amino acid residues or fewer, typically 800 amino acid residues or fewer.
[0159] Suitably the Rv1196 related antigen will comprise, such as consist of, a sequence having at least 70% identity to SEQ ID No: 1, such as at least 80%, in particular at least 90%, especially at least 95%, for example at least 98%, such as at least 99%.
[0160] A specific example of an Rv1196 related antigen is Rv1196 from Mycobacterium tuberculosis strain H37Rv, as provided in SEQ ID No: 1. Consequently, in one embodiment of the invention the Rv1196 related antigen is a protein comprising SEQ ID No: 1. In a second embodiment of the invention the Rv1196 related antigen is a protein consisting of SEQ ID No: 1.
[0161] A further example of an Rv1196 related antigen is Rv1196 from Mycobacterium tuberculosis strain F11. In one embodiment of the invention the Rv1196 related antigen is a protein comprising SEQ ID No: 2. In a second embodiment of the invention the Rv1196 related antigen is a protein consisting of SEQ ID No: 2.
[0162] Typical Rv1196 related antigens will comprise (such as consist of) an immunogenic derivative of SEQ ID No: 1 having a small number of deletions, insertions and/or substitutions. Examples are those having deletions of up to 5 residues at 0-5 locations, insertions of up to 5 residues at 0-5 five locations and substitution of up to 20 residues.
[0163] Other immunogenic derivatives of Rv1196 are those comprising (such as consisting of) a fragment of SEQ ID No: 1 which is at least 200 amino acids in length, such as at least 250 amino acids in length, in particular at least 300 amino acids in length, especially at least 350 amino acids in length.
[0164] Additional immunogenic derivatives of Rv1196 are those comprising, such as consisting of, a fragment of SEQ ID No: 2 which is at least 200 amino acids in length, such as at least 250 amino acids in length, in particular at least 300 amino acids in length, especially at least 350 amino acids in length.
[0165] Rv1196 related antigens may be prepared by methods previously described (e.g. Dillon et al Infection and Immunity 1999 67 (6): 2941-2950; WO2006/117240), those provided in the Examples, or methods analogous thereto.
[0166] In one embodiment the polypeptide antigen and the encoded antigen are Rv0125 related antigens. The term `Rv0125 related antigen` refers to the Rv0125 protein provided in SEQ ID No: 3 or an immunogenic derivative thereof. As used herein the term "derivative" refers to an antigen that is modified relative to the reference sequence. Immunogenic derivatives are sufficiently similar to the reference sequence to substantially retain the immunogenic properties of the reference sequence and remain capable of allowing an immune response to be raised against the reference sequence. An immunogenic derivative may, for example, comprise a modified version of the reference sequence or alternatively may consist of a modified version of the reference sequence.
[0167] The Rv0125 related antigen may for example contain 2500 amino acid residues or fewer, such 1500 amino acid residues or fewer, in particular 1200 amino acid residues or fewer, especially 1000 amino acid residues or fewer, typically 800 amino acid residues or fewer.
[0168] Suitably the Rv0125 related antigen will comprise, such as consist of, a sequence having at least 70% identity to SEQ ID No: 3, such as at least 80%, in particular at least 90%, especially at least 95%, for example at least 98%, such as at least 99%.
[0169] A specific example of an Rv0125 related antigen is Rv0125 from Mycobacterium tuberculosis strain H37Rv, as provided in SEQ ID No: 3. Consequently, in one embodiment of the invention the Rv0125 related antigen is a protein comprising SEQ ID No: 3. In a second embodiment of the invention the Rv0125 related antigen is a protein consisting of SEQ ID No: 3.
[0170] Typical Rv0125 related antigens will comprise (such as consist of) an immunogenic derivative of SEQ ID No: 3 having a small number of deletions, insertions and/or substitutions. Examples are those having deletions of up to 5 residues at 0-5 locations, insertions of up to 5 residues at 0-5 five locations and substitution of up to 20 residues.
[0171] Other immunogenic derivatives of Rv0125 are those comprising (such as consisting of) a fragment of SEQ ID No: 3 which is at least 150 amino acids in length, such as at least 200 amino acids in length, in particular at least 250 amino acids in length, especially at least 300 amino acids in length. Particular immunogenic derivatives of Rv0125 are those comprising (such as consisting of) the fragment of SEQ ID No: 3 corresponding to residues 1-195 of SEQ ID No: 3. Further immunogenic derivatives of Rv0125 are those comprising (such as consisting of) the fragment of SEQ ID No: 3 corresponding to residues 192-323 of SEQ ID No: 3.
[0172] Particularly preferred Rv0125 related antigens are derivatives of SEQ ID No: 3 wherein at least one (for example one, two or even all three) of the catalytic triad have been substituted or deleted, such that the protease activity has been reduced and the protein more easily produced--the catalytic serine residue may be deleted or substituted (e.g. substituted with alanine) and/or the catalytic histidine residue may be deleted or substituted and/or substituted the catalytic aspartic acid residue may be deleted or substituted. Especially of interest are derivatives of SEQ ID No: 3 wherein the catalytic serine residue has been substituted (e.g. substituted with alanine). Also of interest are Rv0125 related antigens which comprise, such as consist of, a sequence having at least 70% identity to SEQ ID No: 3, such as at least 80%, in particular at least 90%, especially at least 95%, for example at least 98%, such as at least 99% and wherein at least one of the catalytic triad have been substituted or deleted or those comprising, such as consisting of, a fragment of SEQ ID No: 3 which is at least 150 amino acids in length, such as at least 200 amino acids in length, in particular at least 250 amino acids in length, especially at least 300 amino acids in length and wherein at least one of the catalytic triad have been substituted or deleted. Further immunogenic derivatives of Rv0125 are those comprising (such as consisting of) the fragment of SEQ ID No: 3 corresponding to residues 192-323 of SEQ ID No: 3 wherein at least one (for example one, two or even all three) of the catalytic triad have been substituted or deleted. Particular immunogenic derivatives of Rv0125 are those comprising (such as consisting of) the fragment of SEQ ID No: 3 corresponding to residues 1-195 of SEQ ID No: 3 wherein the catalytic serine residue (position 176 of SEQ ID No: 3) has been substituted (e.g. substituted with alanine).
[0173] In certain embodiments the polypeptide antigen and the encoded antigen are Rv1196 and Rv0125 related antigens, such as M72 related antigens. Particular derivatives of the M72 protein include those with additional His residues at the N-terminus (e.g. two His residues, as provided in SEQ ID No: 4; or a polyhistidine tag of five or particularly six His residues, which may be used for nickel affinity purification). Mtb72f which contains the original serine residue that has been mutated in M72, is a further derivative of M72, as are Mtb72f proteins with additional His residues at the N-terminus (e.g. two His residues; or a polyhistidine tag of five or particularly six His residues, which may be used for nickel affinity purification).
[0174] Nevertheless, the skilled person recognises that in some embodiments two distinct polypeptides, one being a Rv1196 related antigens and one being a Rv0125 related antigen may be provided within a composition. In such cases it will be recognised that the previously stated exclusions in respect of adenoviruses encoding a Rv1196 related antigen and adenoviruses encoding a Rv0125 related antigen may both be applied to the composition mutatis mutandis. Equally, the previously stated exclusions in respect of contemporaneous administration of adenoviruses encoding a Rv1196 related antigen and adenoviruses encoding a Rv0125 related antigen may both be applied mutatis mutandis.
[0175] Also in some embodiments a single adenovirus may encode two distinct polypeptides, one being a Rv1196 related antigen and one being a Rv0125 related antigen. In such cases it will be recognised that the previously stated exclusions in respect of a polypeptide Rv1196 related antigen and a polypeptide Rv0125 related antigen may both be applied to the composition mutatis mutandis. Equally, the previously stated exclusions in respect of contemporaneous administration of a polypeptide Rv1196 related antigen and a polypeptide Rv0125 related antigen may both be applied mutatis mutandis.
[0176] Alternatively, two distinct adenovirus constructs may be provided, one encoding an Rv1196 related antigen and one encoding an Rv0125 related antigen. In such cases it will be recognised that the previously stated exclusions in respect of a polypeptide Rv1196 related antigen and a polypeptide Rv0125 related antigen may both be applied to the composition mutatis mutandis. Equally, the previously stated exclusions in respect of contemporaneous administration of a polypeptide Rv1196 related antigen and a polypeptide Rv0125 related antigen may both be applied mutatis mutandis.
[0177] Suitably an M72 related antigen will comprise, such as consist of, a sequence having at least 70% identity to SEQ ID No. 6, such as at least 80%, in particular at least 90%, especially at least 95%, such as at least 98%, for example at least 99%.
[0178] Typical M72 related antigens will comprise, such as consist of, a derivative of SEQ ID No: 6 having a small number of deletions, insertions and/or substitutions. Examples are those having deletions of up to 5 residues at 0-5 locations, insertions of up to 5 residues at 0-5 five locations and substitution of up to 20 residues.
[0179] Other derivatives of M72 are those comprising, such as consisting of, a fragment of SEQ ID No: 6 which is at least 450 amino acids in length, such as at least 500 amino acids in length, such as at least 550 amino acids in length, such as at least 600 amino acids in length, such as at least 650 amino acids in length or at least 700 amino acids in length. As M72 is a fusion protein derived from two individual antigens, any fragment of at least 450 residues will comprise a plurality of epitopes from the full length sequence (Skeiky et al J. Immunol. 2004 172:7618-7628; Skeiky Infect. Immun. 1999 67 (8):3998-4007; Dillon Infect. Immun. 1999 67 (6):2941-2950;).
[0180] In particular embodiments the M72 related antigen will comprise residues 2-723 of SEQ ID No. 6, for example comprise (or consist of) SEQ ID No. 6.
[0181] In one embodiment, the polypeptide antigen corresponds to SEQ ID No. 4 and the encoded antigen to SEQ ID No. 6.
[0182] M72 related antigens may be prepared by methods previously described (WO2006/117240) or methods analogous thereto.
[0183] The polypeptide antigen may be the same as or may be similar to the encoded antigen. In one embodiment the polypeptide antigen is the same as the encoded antigen.
[0184] Suitably the polypeptide antigen is similar to the encoded antigen. For example, the encoded antigen may have 70% identity, such as at least 80% identity, suitably at least 90% identity, in particular at least 95% identity to the polypeptide antigen. Alternatively, the encoded antigen may comprise a fragment of at least 100 amino acid residues, such as at least 200 amino acids residues, suitably at least 300 amino acid residues of the polypeptide antigen. In some cases, the encoded antigen comprises a fragment of at least 400 amino acid residues, in particular 500 amino acid residues, such as at least 600 and suitably at least 700 residues of the polypeptide mycobacterial antigen.
[0185] The polypeptide antigen and the adenovirus may be provided in the form of immunogenic compositions which comprise one or more further antigenic components.
[0186] Additional antigenic components may be intended to strengthen or complement the immune responses solicited in the field of tuberculosis prevention and therapy or additional antigens could be associated with other pathogens and are intended for co-administration for reasons of convenience. Where a number of antigenic components are present within a composition, these may be provided in the form of individual polypeptides or fusion proteins. In some circumstances additional antigenic components may be provided as a polynucleotide (or polynucleotides) encoding one or more polypeptides.
[0187] Typically for administration to humans compositions containing a polypeptide Rv1196 related antigen will comprise between 1 ug and 100 ug of Rv1196 related antigen, such as between 1 ug and 50 ug per dose. Suitably between 1 ug and 50 ug of Rv1196 related antigen (such as between 5 ug and 50 ug), especially between 1 ug and 20 ug (such as between 5 ug and 20 ug) and in particular around or exactly 10 ug is provided.
[0188] Typically for administration to humans compositions containing a polypeptide Rv0125 related antigen will comprise between 1 ug and 100 ug of Rv0125 related antigen, such as between 1 ug and 50 ug per dose. Suitably between 1 ug and 50 ug of Rv0125 related antigen (such as between 5 ug and 50 ug), especially between 1 ug and 20 ug (such as between 5 ug and 20 ug) and in particular around or exactly 10 ug is provided.
[0189] Typically for administration to humans compositions containing a polypeptide M72 related antigen will comprise between 1 ug and 100 ug of M72 related antigen, such as between 1 ug and 50 ug per dose. Suitably between 1 ug and 50 ug of M72 related antigen (such as between 5 ug and 50 ug), especially between 1 ug and 20 ug (such as between 5 ug and 20 ug) and in particular around or exactly 10 ug is provided.
[0190] Generally, a polypeptide of use in the invention (if found in nature) will be an isolated polypeptide (i.e. separated from those components with which it may usually be found). For example, a naturally-occurring protein is isolated if it is separated from some or all of the coexisting materials in the natural system. Preferably, such polypeptides are at least about 90% pure, more preferably at least about 95% pure and most preferably at least about 99% pure. A polynucleotide is considered to be isolated if, for example, it is cloned into a vector that is not a part of the natural environment.
Adjuvants of Use in the Invention
[0191] As described above, in one aspect of the invention, polypeptide antigen is provided in an immunogenic composition which comprises an adjuvant. Suitably the adjuvant comprises a TLR agonist and/or an immunologically active saponin.
[0192] In some embodiments the adjuvant may comprise aluminium hydroxide or aluminium phosphate.
[0193] Thus, in one embodiment, the adjuvant comprises a TLR agonist. In another embodiment, the adjuvant comprises an immunologically active saponin. In yet another embodiment, the adjuvant comprises a TLR agonist and an immunologically active saponin.
[0194] The adjuvant may comprise a TLR agonist and a saponin in a liposomal formulation. The ratio of TLR agonist to saponin may be between 5:1 and 1:5 (w/w), suitably between 2:1 and 1:2, typically around 1:1.
[0195] The use of TLR agonists in adjuvants is well-known in the art and has been reviewed e.g. by Lahiri et al. (2008) Vaccine 26:6777. TLRs that can be stimulated to achieve an adjuvant effect include TLR2, TLR4, TLR5, TLR7, TLR8 and TLR9. TLR2, TLR4, TLR7 and TLR8 agonists, particularly TLR4 agonists are preferred.
[0196] Suitable TLR4 agonists include lipopolysaccharides, such as monophosphoryl lipid A (MPL) and 3-O-deacylated monophosphoryl lipid A (3D-MPL). U.S. Pat. No. 4,436,727 discloses MPL and its manufacture. U.S. Pat. No. 4,912,094 and reexamination certificate U.S. Pat. No. 4,912,094 B1 discloses 3D-MPL and a method for its manufacture. Another TLR4 agonist is glucopyranosyl lipid adjuvant (GLA), a synthetic lipid A-like molecule (see, e.g. Fox et al. (2012) Clin. Vaccine Immunol 19:1633). In a further embodiment, the TLR4 agonist may be a synthetic TLR4 agonist such as a synthetic disaccharide molecule, similar in structure to MPL and 3D-MPL or may be synthetic monosaccharide molecules, such as the aminoalkyl glucosaminide phosphate (AGP) compounds disclosed in, for example, WO9850399, WO0134617, WO0212258, WO3065806, WO04062599, WO06016997, WO0612425, WO03066065, and WO0190129. Such molecules have also been described in the scientific and patent literature as lipid A mimetics. Lipid A mimetics suitably share some functional and/or structural activity with lipid A, and in one aspect are recognised by TLR4 receptors. AGPs as described herein are sometimes referred to as lipid A mimetics in the art. In a preferred embodiment, the TLR4 agonist is 3D-MPL. TLR4 agonists, such as 3-O-deacylated monophosphoryl lipid A (3D-MPL), and their use as adjuvants in vaccines has e.g. been described in WO 96/33739 and WO2007/068907 and reviewed in Alving et al. (2012) Curr Opin Immunol 24:310.
[0197] The adjuvant may comprise an immunologically active saponin, such as an immunologically active saponin fraction, such as QS21.
[0198] Adjuvants comprising saponins have been described in the art. Saponins are described in: Lacaille-Dubois and Wagner (1996) A review of the biological and pharmacological activities of saponins. Phytomedicine vol 2:363. Saponins are known as adjuvants in vaccines. For example, Quil A (derived from the bark of the South American tree Quillaja Saponaria Molina), was described by Dalsgaard et al. in 1974 ("Saponin adjuvants", Archiv. fur die gesamte Virusforschung, Vol. 44, Springer Verlag, Berlin, 243) to have adjuvant activity. Purified fractions of Quil A have been isolated by HPLC which retain adjuvant activity without the toxicity associated with Quil A (Kensil et al. (1991) J. Immunol. 146: 431. Quil A fractions are also described in U.S. Pat. No. 5,057,540 and "Saponins as vaccine adjuvants", Kensil, C. R., Crit Rev Ther Drug Carrier Syst, 1996, 12 (1-2):1-55.
[0199] Two such fractions, suitable for use in the present invention, are QS7 and QS21 (also known as QA-7 and QA-21). QS21 is a preferred immunologically active saponin fraction for use in the present invention. QS21 has been reviewed in Kensil (2000) In O'Hagan: Vaccine Adjuvants: preparation methods and research protocols. Homana Press, Totowa, N.J., Chapter 15. Particulate adjuvant systems comprising fractions of Quil A, such as QS21 and QS7, are e.g. described in WO 96/33739, WO 96/11711 and WO2007/068907.
[0200] In addition to the other components, the adjuvant preferably comprises a sterol. The presence of a sterol may further reduce reactogenicity of compositions comprising saponins, see e.g. EP0822831. Suitable sterols include beta-sitosterol, stigmasterol, ergosterol, ergocalciferol and cholesterol. Cholesterol is particularly suitable. Suitably, the immunologically active saponin fraction is QS21 and the ratio of QS21:sterol is from 1:100 to 1:1 w/w, such as from 1:10 to 1:1 w/w, e.g. from 1:5 to 1:1 w/w.
[0201] In a preferred embodiment of the methods of the invention, the TLR4 agonist is 3D-MPL and the immunologically active saponin is QS21.
[0202] In some embodiments, the adjuvant is presented in the form of an oil-in-water emulsion, e.g. comprising squalene, alpha-tocopherol and a surfactant (see e.g. WO95/17210) or in the form of a liposome. A liposomal presentation is preferred.
[0203] The term "liposome" when used herein refers to uni- or multilamellar (particularly 2, 3, 4, 5, 6, 7, 8, 9, or 10 lamellar depending on the number of lipid membranes formed) lipid structures enclosing an aqueous interior. Liposomes and liposome formulations are well known in the art. Liposomal presentations are e.g. described in WO96/33739 and WO2007/068907. Lipids which are capable of forming liposomes include all substances having fatty or fat-like properties. Lipids which can make up the lipids in the liposomes may be selected from the group comprising glycerides, glycerophospholipides, glycerophosphinolipids, glycerophosphonolipids, sulfolipids, sphingolipids, phospholipids, isoprenolides, steroids, stearines, sterols, archeolipids, synthetic cationic lipids and carbohydrate containing lipids. In a particular embodiment of the invention the liposomes comprise a phospholipid. Suitable phospholipids include (but are not limited to): phosphocholine (PC) which is an intermediate in the synthesis of phosphatidylcholine; natural phospholipid derivates: egg phosphocholine, egg phosphocholine, soy phosphocholine, hydrogenated soy phosphocholine, sphingomyelin as natural phospholipids; and synthetic phospholipid derivates: phosphocholine (didecanoyl-L-a-phosphatidylcholine [DDPC], dilauroylphosphatidylcholine [DLPC], dimyristoylphosphatidylcholine [DMPC], dipalmitoyl phosphatidylcholine [DPPC], Distearoyl phosphatidylcholine [DSPC], Dioleoyl phosphatidylcholine, [DOPC], 1-palmitoyl, 2-oleoylphosphatidylcholine [POPC], Dielaidoyl phosphatidylcholine [DEPC]), phosphoglycerol (1,2-Dimyristoyl-sn-glycero-3-phosphoglycerol [DMPG], 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol [DPPG], 1,2-distearoyl-sn-glycero-3-phosphoglycerol [DSPG], 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol [POPG]), phosphatidic acid (1,2-dimyristoyl-sn-glycero-3-phosphatidic acid [DMPA], dipalmitoyl phosphatidic acid [DPPA], distearoyl-phosphatidic acid [DSPA]), phosphoethanolamine (1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine [DMPE], 1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine [DPPE], 1,2-distearoyl-sn-glycero-3-phosphoethanolamine [DSPE], 1,2-Dioleoyl-sn-Glycero-3-Phosphoethanolamine [DOPE]), phoshoserine, polyethylene glycol [PEG] phospholipid.
[0204] Liposome size may vary from 30 nm to several um depending on the phospholipid composition and the method used for their preparation. In particular embodiments of the invention, the liposome size will be in the range of 50 nm to 500 nm and in further embodiments 50 nm to 200 nm. Dynamic laser light scattering is a method used to measure the size of liposomes well known to those skilled in the art.
[0205] In a particularly suitable embodiment, liposomes used in the invention comprise DOPC and a sterol, in particular cholesterol. Thus, in a particular embodiment, compositions of the invention comprise QS21 in any amount described herein in the form of a liposome, wherein said liposome comprises DOPC and a sterol, in particular cholesterol.
[0206] Preferably, the adjuvant comprises 3D-MPL and QS21 in a liposomal formulation.
[0207] The adjuvant may comprise 1 to 100 micrograms of TLR4 agonist per dose.
[0208] The adjuvant may comprise 1 to 100 micrograms of immunologically active saponin per dose.
[0209] In one embodiment for human use, the adjuvant comprises between 12.5 and 75 micrograms of 3D-MPL and between 12.5 and 75 micrograms of QS21 per dose in a liposomal formulation.
[0210] In another embodiment, the adjuvant comprises between 12.5 and 37.5, such as between 20 and 30 micrograms (for example about or exactly 25 micrograms), of 3D-MPL and between 12.5 and 37.5, such as between 20 and 30 micrograms (for example about or exactly 25 micrograms) of QS21 in a liposomal formulation per dose. Suitably the amount of 3D-MPL is the same as the amount of QS21.
[0211] The polypeptide or adenovirus should be presented in a pharmaceutically acceptable form, appropriate to the intended delivery route. Solutions should have a pharmaceutically acceptable osmolality to avoid cell distortion or lysis. A pharmaceutically acceptable osmolality will generally mean that solutions will have an osmolality which is approximately isotonic or mildly hypertonic. Suitably the immunogenic compositions of the present invention will have an osmolality in the range of 250 to 750 mOsm/kg, for example, the osmolality may be in the range of 250 to 550 mOsm/kg, such as in the range of 280 to 500 mOsm/kg. Osmolality may be measured according to techniques known in the art, such as by the use of a commercially available osmometer, for example the Advanced.RTM. Model 2020 available from Advanced Instruments Inc. (USA). An "isotonicity agent" is a compound that is physiologically tolerated and imparts a suitable tonicity to a formulation (e.g. immunogenic compositions of the invention) to prevent the net flow of water across cell membranes that are in contact with the formulation. Aqueous adjuvant compositions are known which contain 100 mM sodium chloride or more, for example adjuvant system A (ASA) in WO 2005/112991 and WO2008/142133 or the liposomal adjuvants disclosed in WO2007/068907.
[0212] In some embodiments, the isotonicity agent used for the composition is a salt. In other embodiments, however, the composition comprises a non-ionic isotonicity agent and the concentration of sodium chloride or the ionic strength in the composition is less than 100 mM, such as less than 80 mM, e.g. less than 30 mM, such as less 10 mM or less than 5 mM. The composition may comprise a non-ionic isotonicity agent and conductivity of the composition is less than 5 mS/cm, such as less than 4 mS/cm. In a preferred embodiment, the non-ionic isotonicity agent is a polyol, such as sorbitol. The concentration of sorbitol may e.g. between about 3% and about 15% (w/v), such as between about 4% and about 10% (w/v). Adjuvants comprising an immunologically active saponin fraction and a TLR4 agonist wherein the isotonicity agent is salt or a polyol have been described in WO2012/080369 and WO2012/080370 which are incorporated herein by reference.
[0213] The pH of the immunogenic compositions should be suitable for administration. Typically the pH will be in the range 6.0 to 9.0, such as 7.0 to 9.0, especially 7.25 to 8.75, such as 7.5 to 8.5, in particular pH 7.75 to 8.25. A pH of about 8.0 is of particular interest.
[0214] For liquid compositions administered parenterally, the volume of the composition will typically be in the region of 50 ul to 2 ml (depending on the specific route). A volume of 400-600 ul, such as around 500 ul is typically used, in particular for administration by the intramuscular route.
[0215] The compositions will generally be sterile.
Adenoviral Vectors
[0216] Adenovirus has been widely used for gene transfer applications due to its ability to achieve highly efficient gene transfer in a variety of target tissues and large transgene capacity. Adenoviral vectors of use in the present invention may be derived from a range of mammalian hosts. Over 100 distinct serotypes of adenovirus have been isolated which infect various mammalian species. These adenoviral serotypes have been categorised into six subgenera (A-F; B is subdivided into B1 and B2) according to sequence homology and on their ability to agglutinate red blood cells (Tatsis and Ertl Molecular Therapy (2004) 10:616-629).
[0217] Examples of human-derived adenoviruses are Ad1, Ad2, Ad4, Ad5, Ad6, Ad11, Ad 24, Ad34 and Ad35. Although Ad5-based vectors have been used extensively in a number of gene therapy trials, there may be limitations on the use of Ad5 and other human group C adenoviral vectors due to preexisting immunity in the general population due to natural infection. Ad5 and other human group C members tend to be among the most seroprevalent serotypes. Additionally, immunity to existing vectors may develop as a result of exposure to the vector during treatment. These types of preexisting or developed immunity to seroprevalent vectors may limit the effectiveness of gene therapy or vaccination efforts. Alternative adenovirus serotypes, thus constitute very important targets in the pursuit of gene delivery systems capable of evading the host immune response.
[0218] The adenoviral vector of use in the present invention is derived from a non-human simian adenovirus. Numerous adenoviruses have been isolated from non-human simians such as chimpanzees, bonobos, rhesus macaques and gorillas, and vectors derived from these adenoviruses induce strong immune responses to transgenes encoded by these vectors (Colloca et al. (2012) Sci. Transl. Med. 4:1-9; Roy et al. (2004) Virol. 324: 361-372; Roy et al. (2010) J. of Gene Med. 13:17-25). Certain advantages of vectors based on non-human simian adenoviruses include the relative lack of cross-neutralising antibodies to these adenoviruses in the target population. For example, cross-reaction of certain chimpanzee adenoviruses with pre-existing neutralizing antibody responses is only present in 2% of the target population compared with 35% in the case of certain candidate human adenovirus vectors.
[0219] Specifically, the adenoviral vector is derived from a non-human simian adenovirus, in particular a chimpanzee adenovirus such as ChAd3, ChAd63, ChAd83, ChAd155, Pan 5, Pan 6, Pan 7 (also referred to as C7) or Pan 9. Examples of such strains are described in WO03/000283, WO2005/071093, WO2010/086189 and GB1510357.5 and are also available from the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209, and other sources. Alternatively, adenoviral vectors may be derived from non-human simian adenoviruses isolated from bonobos, such as PanAd1, PanAd2 or PanAd3. Examples of such vectors described herein can be found for example in WO2005/071093 and WO2010/086189. Adenoviral vectors may also be derived from adenoviruses isolated from gorillas as described in WO2013/52799, WO2013/52811 and WO2013/52832.
[0220] Certain adenoviral vectors may demonstrate one or more following improved characteristics over other vectors: higher productivity, improved immunogenicity and increased transgene expression.
[0221] In one embodiment the adenoviral vector is a non-human simian adenovirus containing at least a penton selected from SEQ ID No: 10 or SEQ ID No: 20, a hexon selected from SEQ ID No: 11 or SEQ ID No: 21 or a fibre selected from SEQ ID No: 12 or SEQ ID No: 22, in particular a penton and hexon, penton and fibre, or hexon and fibre, such as a penton, hexon and fibre. In certain examples the adenoviral vector is a non-human simian adenovirus containing at least the penton of SEQ ID No: 10, the hexon of SEQ ID No: 11 or the fibre of SEQ ID No: 12, in particular a penton and hexon, penton and fibre, or hexon and fibre. In other examples the adenoviral vector is a non-human simian adenovirus containing at least the penton of SEQ ID No: 20, the hexon of SEQ ID No: 21 or the fibre of SEQ ID No: 22, in particular a penton and hexon, penton and fibre, or hexon and fibre.
[0222] In an embodiment the adenoviral vector is derived from as ChAd3 and contains at least a penton (SEQ ID No: 10), hexon (SEQ ID No: 11) and fibre (SEQ ID No: 12) therefrom.
[0223] In one embodiment the adenoviral vector is derived from as ChAd155 and contains a penton (SEQ ID No: 20), hexon (SEQ ID No: 21) and fibre (SEQ ID No: 22) therefrom.
[0224] In one embodiment the adenoviral vector is a non-human simian adenovirus containing at least the penton of SEQ ID No: 15, the hexon of SEQ ID No: 16 or the fibre of SEQ ID No: 17, in particular a penton and hexon, penton and fibre, or hexon and fibre. In some examples the adenoviral vector is derived from as ChAd63 containing at least the penton of SEQ ID No: 15, the hexon of SEQ ID No: 16 and the fibre of SEQ ID No: 17.
Adenoviral Vector Structure
[0225] Adenoviruses have a characteristic morphology with an icosahedral capsid comprising three major proteins, hexon (II), penton base (III) and a knobbed fiber (IV), along with a number of other minor proteins, VI, VIII, IX, IIIa and IVa2. The hexon accounts for the majority of the structural components of the capsid, which consists of 240 trimeric hexon capsomeres and 12 penton bases. The hexon has three conserved double barrels, while the top has three towers, each tower containing a loop from each subunit that forms most of the capsid. The base of hexon is highly conserved between adenoviral serotypes, while the surface loops are variable (Tatsis and Ertl Molecular Therapy (2004) 10:616-629). Penton is another adenoviral capsid protein that forms a pentameric base to which fiber attaches. The trimeric fiber protein protrudes from the penton base at each of the 12 vertices of the capsid and is a knobbed rod-like structure. The primary role of the fibre protein is the tethering of the viral capsid to the cell surface via the interaction of the knob region with a cellular receptor, and variations in the flexible shaft as well as knob regions of fiber are characteristic of the different serotypes (Nicklin et al Molecular Therapy 2005 12:384-393).
[0226] The adenoviral genome is well characterised. The linear, double-stranded DNA is associated with the highly basic protein VII and a small peptide pX (also termed mu). Another protein, V, is packaged with this DNA-protein complex and provides a structural link to the capsid via protein VI. There is general conservation in the overall organization of the adenoviral genome with respect to specific open reading frames being similarly positioned, e.g. the location of the E1A, E1 B, E2A, E2B, E3, E4, L1, L2, L3, L4 and L5 genes of each virus. Each extremity of the adenoviral genome comprises a sequence known as an inverted terminal repeat (ITR), which is necessary for viral replication. The 5' end of the adenoviral genome contains the 5' cis-elements necessary for packaging and replication; i.e., the 5' ITR sequences (which function as origins of replication) and the native 5' packaging enhancer domains (that contain sequences necessary for packaging linear Ad genomes and enhancer elements for the E1 promoter). The 3' end of the adenoviral genome includes the 3' cis-elements (including the ITRs) necessary for packaging and encapsidation. The virus also comprises a virus-encoded protease, which is necessary for processing some of the structural proteins required to produce infectious virions. The structure of the adenoviral genome is described on the basis of the order in which the viral genes are expressed following host cell transduction. More specifically, the viral genes are referred to as early (E) or late (L) genes according to whether transcription occurs prior to or after onset of DNA replication. In the early phase of transduction, the E1A, E1B, E2A, E2B, E3 and E4 genes of adenovirus are expressed to prepare the host cell for viral replication. During the late phase of infection, expression of the late genes L1-L5, which encode the structural components of the virus particles, is activated.
Annotation of the ChAd3 wild type sequence (SEQ ID NO: 9) sequence is provided below. CDS (38 total)
[0227] E1A 30.8K
[0228] Start: 589 End: 1544
[0229] Original Location Description:
[0230] join(589..1129, 1243..1544)
[0231] E1A 25.5K
[0232] Start: 589 End: 1544
[0233] Original Location Description:
[0234] join(589..991, 1243. .1544)
[0235] E1B 22K
[0236] Start: 1716 End: 2279
[0237] Original Location Description:
[0238] 1716..2279
[0239] E1B 57K
[0240] Start: 2021 End: 3544
[0241] Original Location Description:
[0242] 2021..3544
[0243] IX
[0244] Start: 3640 End: 4104
[0245] Original Location Description:
[0246] 3640..4104
[0247] IVa2
[0248] Start: 4163 End: 5790 (Complementary)
[0249] Original Location Description:
[0250] complement(4163..5499,5778..5790)
[0251] pol
[0252] Start: 5269 End: 14236 (Complementary)
[0253] Original Location Description:
[0254] complement(5269..8865,14228.. 14236)
[0255] pTP
[0256] Start: 8664 End: 14236 (Complementary)
[0257] Original Location Description:
[0258] complemenl(8664.. 10667,14228.. 14236)
[0259] 48K
[0260] Start: 11120 End: 12379
[0261] Original Location Description:
[0262] 11120..12379
[0263] pIIIa
[0264] Start: 12403 End: 14181
[0265] Original Location Description:
[0266] 12403..14181
[0267] III
[0268] Start: 14273 End: 16054
[0269] Penton
[0270] Original Location Description:
[0271] 14273..16054
[0272] pVII
[0273] Start: 16069 End: 16665
[0274] Original Location Description:
[0275] 16069.. 16665
[0276] V
[0277] Start: 16738 End: 17853
[0278] Original Location Description:
[0279] 16738.. 17853
[0280] pX
[0281] Start: 17878 End: 18123
[0282] Original Location Description:
[0283] 17878..18123
[0284] pVI
[0285] Start: 18219 End: 18974
[0286] Original Location Description:
[0287] 18219..18974
[0288] hexon
[0289] Start: 19086 End: 21968
[0290] Original Location Description:
[0291] 19086..21968
[0292] protease
[0293] Start: 21998 End: 22627
[0294] Original Location Description:
[0295] 21998..22627
[0296] DBP
[0297] Start: 22743 End: 24395 (Complementary)
[0298] Original Location Description:
[0299] complement(22743..24395)
[0300] 92K
[0301] Start: 24445 End: 26940
[0302] Original Location Description:
[0303] 24445..26940
[0304] 22K
[0305] Start: 26630 End: 27229
[0306] Original Location Description:
[0307] 26630..27229
[0308] 33K
[0309] Start: 26630 End: 27551
[0310] Original Location Description:
[0311] join(26630..26966,27169..27551)
[0312] pVIII
[0313] Start: 27626 End: 28309
[0314] Original Location Description:
[0315] 27626..28309
[0316] E3 12K
[0317] Start: 28310 End: 28627
[0318] Original Location Description: 28310..28627
[0319] E3 CR1-alphap0
[0320] Start: 29125 End: 29325
[0321] Original Location Description:
[0322] 29125..29325
[0323] E3gp18K
[0324] Start: 29328 End: 29819
[0325] Original Location Description:
[0326] 29328..29819
[0327] E3 33K
[0328] Start: 29848 End: 30738
[0329] Original Location Description:
[0330] 29848..30738
[0331] E3A 11 K
[0332] Start: 31293 End: 31589
[0333] Original Location Description:
[0334] 31293..31589
[0335] E3 RID alpha
[0336] Start: 31601 End: 31873
[0337] Original Location Description:
[0338] 31601 .31873
[0339] E3 RID beta
[0340] Start: 31876 End: 32274
[0341] Original Location Description:
[0342] 31876..32274
[0343] E3 15K
[0344] Start: 32267 End: 32653
[0345] Original Location Description:
[0346] 32267..32653
[0347] U exon
[0348] Start: 32684 End: 32848 (Complementary)
[0349] Original Location Description:
[0350] complement(32684.. 32848)
[0351] fiber
[0352] Start: 32859 End: 34490
[0353] Original Location Description:
[0354] 32859..34490
[0355] E4 ORF6/7
[0356] Start: 34698 End: 35858 (Complementary)
[0357] Original Location Description:
[0358] complement(34698..34973,35685..35858)
[0359] E4 ORF6
[0360] Start: 34974 End: 35858 (Complementary)
[0361] Original Location Description:
[0362] complement (34974.. 35858)
[0363] E4 ORF4
[0364] Start: 35758 End: 36123 (Complementary)
[0365] Original Location Description:
[0366] complement(35758..36123)
[0367] E4 ORF3
[0368] Start: 36139 End: 36486 (Complementary)
[0369] Original Location Description:
[0370] complement(36139..36486)
[0371] E4 ORF2
[0372] Start: 36483 End: 36875 (Complementary)
[0373] Original Location Description:
[0374] complement(36483..36875)
[0375] E4 ORF1
[0376] Start: 36928 End: 37314 (Complementary)
[0377] Original Location Description:
[0378] complement(36928..37314) Misc. Feature (3 total)
[0379] VA RNA I
[0380] Start: 10693 End: 10860
[0381] Original Location Description:
[0382] 10693.. 10860
[0383] VA II
[0384] Start: 10927 End: 11102
[0385] Original Location Description:
[0386] 10927.11102
[0387] E3 deletion - 5'
[0388] Start: 28642 End: 28647
[0389] Original Location Description:
[0390] 28642..28647 Annotation of the ChAd63 wild type sequence (SEQ ID NO: 14) sequence is provided below.
TABLE-US-00001
[0390] LOCUS ChAd63 35994 bp DNA linear 27-JUL-2015 DEFINITION Chimp adenovirus 163, complete genome. COMMENT Annotation according to alignment of ChAd63 against the human Adenovirus 4 reference strain NC 003266 FEATURES Location/Qualifiers source 1..35994 /organism=''Chimpanzee adenovirus 63'' /mol_type=''genomic DNA'' /acronym=''ChAd63'' repeat_region 1..129 /standard_name=''ITR'' /rpt_type=inverted gene 479..1501 /gene=''E1A'' regulatory 479..484 /regulatory_class=''TATA_box'' /gene=''E1A'' CDS join(576..1143,1229..1437) /gene=''E1A'' /product=''control protein E1A'' /translation=''MRHLRDLPGNVFLATGNEILELVVDAMMGDDPPEPPTPFEAPSL YDLYDLEVDVPENDPNEEAVNDLFSDAALLAAEQANTDSGSDSDSSLHTPRPGRGEKK IPELKGEELDLRCYEECLPPSDDEEDEEAIRAAASEGVKVAGESFSLDCPTLPGHGCK SCEFHRMNTGDKNVMCALCYMRAYNHCVYSPVSDVDETPTSECISSPPEIGEEPPEDI IHRPVAVRVTGRRAAVESLDDLLQGGDEPLDLCTRKRPRH'' intron 1144..1228 /gene=''E1A'' regulatory 1495..1501 /regulatory_class=''polyA_signal_sequence'' /gene=''E1A'' gene 1555..3953 /gene=''E1B'' regulatory 1555..1664 /regulatory_class=''TATA_box'' /gene=''E1B'' CDS 1601..2179 /gene=''E1B'' /codon_start=1 /product=''control protein E1B 19K'' /translation=''MEIWTVLEDFHQTRQLLENSSAEVSYLWRFCFGGPLAKLVYRAK QDYKDQFEDILRECPGIFDSLNLGHQSHFNQSILRALDFSTPGRTTAAVAFFAFILDK WSQETHFSRDYRLDCLAVALWRTWRCQRLNAISGYLPVQPVDTLRILSLQSPQEHQRR QQPQQEQQQEEEEDREENLRAGLDPPVAEEEE'' CDS 1906..3420 /gene=''E1B'' /codon_start=1 /product=''control protein E1B 55K'' /translation=''MESRNPFQQGLPSGLLSSSFVENMEVPAPECNLRLLASTAGRHA EDPESPVTPGTPTPPAAAAGAAARGGGGPRREPESRSGPSGGGGGGVADLFPELRRVL TRSSSGRERGIKRERHEETSHRTELTVSLMSRRRPESVWWHEVQSQGIDEVSVMHEKY SLEQVKTCWLEPEDDWEVAIRNYAKLALKPDKKYKITKLINIRNSCYISGNGAEVEIS TQERAAFRCOMMNMYPGVVGMEGVTFMNTRFRGDGYNGVVFMANTKLTVHGCSFFGFN NMCIEAWGSVSVRGCSFSANWMGVVGRTKSVVSVKKCLFERCHLGVMSEGEAKVKHCA STETGCFVLIKGNAKVKHNMICGASDERGYQMLTCAGGNSHMLATVHVASHPRKTWPE FEHNVMTRCNVHLGSRRGMFMPYQCNMQFVKVLLEPDAMSRVSLTGVFDMNVELWKIL RYDESKTRCRACECGGKHARLQPVCVEVTEDLRPDHLVLSCNGTEFGSSGEESD'' gene 3454..3953 /gene=''IX'' regulatory 3454..3459 /regulatory_class=''TATA_box'' /gene=''IX'' CDS 3505..3933 /gene=''IX'' /product=''capsid protein IX'' /translation=''MSGSASFEGGVFSPYLTGRLPSWAGVRQNVMGSTVDGRPVQPAN SSTLTYATLSSSSVDAAAAAAAASAASAVRGMALGAGYYSSLVANSSSTNNPASLNEE KLLLLMAQLEALTQRLGELTQQVAQLQAETRAAVATVKTK'' regulatory 3929..3934 /regulatory_class=''polyA_signal_sequence'' /note=''E1B, IX'' regulatory 3944..3949 /regulatory_class=''polyA_signal_sequence'' /note=''E1B, IX'' regulatory 3948..3953 /regulatory_class=''polyA_signal_sequence'' /note=''E1B, IX'' gene complement(3992..26364) /gene=''E2B'' gene complement(3992..5735) /gene=''IVa2'' regulatory complement(3992..3997) /regulatory_class=''polyA_signal_sequence'' /note=''IVa2, E2B'' CDS complement(join(3993..5326,5605..5617)) /gene=''IVa2'' /product=''encapsidation protein IVa2'' /translation=''METRGRRPGAVLDQPDEPEAHPRKRPARRAPLHRDGDHADADPA TLEGPDPGLAGRPSPGALLPQSPQPAKRGGLLDRDALEHITELWDRLELLQQTLSKMP MADGLKPLKNFASLQELLSLGGERLLAELVRENMHVREMMNEVAPLLREDGSCLSLNY HLQPVIGVIYGPTGCGKSQLLRNLLSAQLISPAPETVFFIAPQVDMIPPSELKAWEMQ ICEGNYAPGIEGTFVPQSGTLRPKFIKMAYDELTQDHNYDVSDPRNVFAQAAAHGPIA IIMDECMENLGGHKGVSKFFHAFPSKLHDKFPKCTGYTVLVVLHNMNPRRDLGGNIAN LKIQAKMHLISPRMHPSQLNRFVNTYTKGLPVAISLLLKDIVQHHALRPCYDWVIYNT TPEHEALQWSYLHPRDGLMPMYLNIQAHLYRVLEKIHRVLNDRDRWSRAYRARKIK'' CDS complement(join(5096..8659,13841..13849)) /gene=''E2B'' /EC_number=''2.7.7.7'' /product=''DNA polymerase'' /translation=''MALVQTHGSRGLHPEASDPGRQPSRRRSRQSSPGAVPEPARARR RRAPAAPASGPRAAPAARRASSPPLLSMEPPPPKKKRGTVVAPQGHGTLQAVDVATNG AVEIKYHLDLPRALEKLLQVNRAPPLPTDLTPQRLRTLDSSGLRALVLALRPVRAEVW TCLPRGLVSMTTIEADDGHADGQDVVQHQMQPPALHCPLKFLVKGTQVQLVQHVHPVQ RCEHCGRLYKHKHECSARRRHFYFHHINSHSSNWWQEIQFFPIGSHPRTERLFLTYDV ETYTWMGSFGKQLVPFMLVMKLSGDPPLVELAHDLALQLKWDRWHGDPRTFYCVTPEK MAVGQQFRQYRDRLQTALAVDLWTSFLSANPHVADWALEQHGLSDPAELTYDELKKLP HVKGRPRFVELYIVGHNINGFDEIVLAAQVINNRAEVPQPFRITRNFMPRAGKILFND VTFALPNPAYKKRTDFQLWEQGGCDDLDFKHQFLKVMVRDTFALTHTSLRKAAQAYAL PVEKGCCAYKAVNQFYMVGSYRADQDGFPLEEYWKDREEFLLNRELWKQKGQLKYDII QETLDYCALDVLVTAELVAKLQDSYAHFIRDSVGLPHAHFNIFQRPTISSNSHAIFRQ IVYRAEKPQRSNLGTGLLAPSHELYDYVRASIRGGRCYPTYIGVLQEPLYVYDICGMY ASALTHPMPWGTPLSPYERALAVRDWQASLDDLGTCISYFDPELLPGIFTVDADPPDE LMLDPLPPFCSRKGGRLCWTNEPLRGEVATSVDLITLHNRGWRVRIVPDELTTVFPEW KCVAREYVQLNIAAKERADKEKNQTMRSIAKLLSNALYGSFATKLDNKKIVFSDQMDE GLMKGVSNGTVNIKSSSFLETDNLSAEVMPAFEREYLPQQLALLDSDPEDSEDEQGPA PFYTPPAGTPGHVAYTYKPITFLDVDEGDMCLHTLEKVDPLVDNDRYPSHVASFVLAW TRAFVSEWAGFLYDEDRGTPLEDRPIKSVYGDTDSLFVTQRGHELMETRGKKRIKKHG GNLVFDPDRPDLTWLVECETVCASCGADAYAPESVFLAPKLYALKSLLCPVCGHTSKG KLRAKGHAAEALNYELMLNCYLADAQGADRERFSTSRMSLKRTLASAQPGAHPFTVTE TTLTRTLRPWKDRTLASLDAHRLVPYSRSRPNPRNEEVCWIEMP'' intron complement(5327..5604) /gene=''IVa2'' gene 5917..33604 /gene=''L5'' gene 5917..27469 /gene=''L4'' gene 5917..21839 /gene=''L3'' gene 5917..17466 /gene=''L2'' gene 5917..13827 /gene=''L1'' regulatory 5917..5922 /regulatory_class=''TATA_box'' /note=''L'' intron 5989..7009 /note=''between L1 and L2 leaders'' intron 7082..9518 /note=''between L2 and L3 leaders'' intron 7082..7850 /note=''between L2 and i leaders; precedes protein 13.6K CDS'' CDS join(7877..8275,9519..9539) /gene=''L1'' /codon_start=1 /product=''protein 13.6K'' /translation=''MRADGEELDLLPPVGGMAVDVMEVEMPTARRALVLVFIQASAVL ATLHGMHVLHELYLGSFDEEFQWAVERWRLHLVLYYVLAIGVAIVCLDGGHADEPARE AGPDLGSDGSESEDEGAQAGAVQGPETLRSQGLRART'' CDS complement(join(8458..10392,13841..13849)) /gene=''E2B'' /product=''terminal protein precursor pTP'' /translation=''MALSIHDCARLTGQTAATMNYFLPLRNIWNRVREFPRASTTAAG ITWMSRYIYGYHRLMLEDLAPGAPATERWPLYRQPPPHFLVGYQYLVRTCNDYIFDTR AYSRLKYHELVRPGHQTVNWSVMANCSYTINTGAYHRFVDFDDFQTTLTQIQQAILAE RVVADLALVQPQRGFGLTRMHGRAGEEEVPVERLMQDYYKDLARCQDHAWGMADRLRI QQAGPKDLVLLATIRRLRTAYFNFITSSIARPPPDQIPEEQETGLSLPCDCDWLEAFV QRFSDPVDLETLRSLRGVPTGQLIRCIVSALSLPNGDPPGGHLEMRGGVFTLRPREDG RAVTETMRRRRGETIERFIDRLPVRRRRRRAPPPPPPPEEEVEEMLVEEEEEEMEEEP PGAFEREVRATIAELIRLLEEELTVSARNSQFFNFAVDFYEAMERLEALGDVSEMPLR RWIMYFFVTEHIATTLNYLYQRLCNYAVFTRHVELNLAQVVMRARDPDGAVVYSRVWN EAGMNAFSQLMGRISNDLAATVERAGRGDLQEEEIEQFMTEIAYQDNSGDVQEILRQA AVNDTEIDSVELSFRFKLTGPVAFTQRRQIQDVNRRVVAHASLLRAQYQNLPARGADV PLPPLPPGPEPPLPPGARPRRRF'' intron complement(8660..26296) /gene=''E2B'' intron 9606..32253 /gene=''L5'' /note=''precedes fiber CDS'' intron 9606..26463 /gene=''L4'' /note=''precedes capsid protein precursor pVIII CDS'' intron 9606..25554 /gene=''L4'' /note=''precedes protein 33K and encapsidation protein 22K CDSs'' intron 9606..23430 /gene=''L4'' /note=''precedes hexon assembly protein 100K CDS'' intron 9606..21177 /gene=''L3'' /note=''precedes protease CDS'' intron 9606..18296 /gene=''L3'' /note=''precedes hexon CDS'' intron 9606..17508 /gene=''L3'' /note=''precedes capsid protein precursor pVI CDS'' intron 9606..17201 /gene=''L2'' /note=''precedes core protein precursor pX CDS'' intron 9606..16132 /gene=''L2'' /note=''precedes core protein V CDS'' intron 9606..15498 /gene=''L2'' /note=''precedes core protein precursor pVII CDS'' intron 9606..13887 /gene=''L2'' /note=''precedes penton base CDS'' intron 9606..12043 /gene=''L1'' /note=''precedes capsid protein precursor pIIIa CDS''
intron 9606..10844 /gene=''L1'' /note=''precedes encapsidation protein 52K CDS'' intron complement(10393..26296) /gene=''E2B'' gene 10426..10585 /gene=''VAI'' misc_RNA 10426..10585 /gene=''VAI'' /product=''virus-associated RNA I'' gene 10648..10818 /gene=''VAII'' misc_RNA 10648..10818 /gene=''VAII'' /product=''virus-associated RNA II'' CDS 10845..12020 /gene=''L1'' /product=''encapsidation protein 52K'' /translation=''MHPVLRQMRPHPPPQPPLPQQQQQPALLPPPQQQQPATTAAAAV SGAGVQYDLALEEGEGLARLGASSPERHPRVQMKRDAREAYVPKQNLFRDRSGEEPEE MRASRFHAGRELRRGLDRKRVLRDEDFEADELTGISPARAHVAAANLVTAYEQTVKEE SNFQKSFNNHVRTLIAREEVTLGLMHLWDLLEAIVQNPTSKPLTAQLFLVVQHSRDNE TFREALLNITEPEGRWLLDLVNILQSIVVQERGLPLSEKLAAINFSVLSLGKYYARKI YKTPYVPIDKEVKIDGFYMRMTLKVLTLSDDLGVYRNDRMHRAVSASRRRELSDQELM HSLQRALTGAGTEGESYFDMGADLRWQPSRRALEAAGGVPYVEEVDDEEEEGEYLED'' CDS 12044..13810 /gene=''L1'' /product=''capsid protein precursor pIIIa'' /translation=''MQQQPPPPPPDPAMRAALQSQPSGINSSDDWTQAMQRIMALTTR NPEAFRQQPQANRLSAILEAVVPSRSNPTHEKVLAIVNALVENKAIRGDEAGLVYNAL LERVARYNSTNVQTNLDRMVTDVREAVSQRERFHRESNLGSMVALNAFLSTQPANVPR GQEDYTNFISALRLMVAEVPQSEVYQSGPDYFFQTSRQGLQTVNLSQAFKNLQGLWGV QAPVGDRATVSSLLTPNSRLLLLLVAPFTDSGSVSRDSYLGYLLNLYREAIGQAHVDE QTYQEITHVSRALGQEDPGNLEATLNFLLTNRSQKIPPQYALSTEEERILRYVQQSVG LFLMQEGATPSAALDMTARNMEPSMYARNRPFINKLMDYLHRAAAMNSDYFTNAILNP HWLPPPGFYTGEYDMPDPNDGFLWDDVDSSVFSPRPTTTTVWKKEGGDRRPSSALSGR AGAAAAVPEAASPFPSLPFSLNSVRSSELGRLTRPRLLGEEEYLNDSLLKPEREKNFP NNGIESLVDKMSRWKTYAHEHRDEPRASSAGTRRRQRHDRQRGLVWDDEDSADDSSVL DLGGSGGGNPFAHLRPRIGRLM'' regulatory 13822..13827 /regulatory_class=''polyA_signal_sequence'' /gene=''L1'' CDS 13889..15511 /gene=''L2'' /product=''penton base'' /translation=''MMRRVYPEGPPPSYESVMQQAVAAAMQPPLEAPYVPPRYLAPTE GRNSIRYSELAPLYDTTRLYLVDNKSADIASLNYQNDHSNFLTTVVQNNDFTPTEAST QTINFDERSRWGGQLKTIMHTNMPNVNEFMYSNKFKARVMVSRKTPNGVTVGDDYDGS QDELTYEWVEFELPEGNFSVTMTIDLMNNAIIDNYLAVGRQNGVLESDIGVKFDTRNF RLGWDPVTELVMPGVYTNEAFHPDIVLLPGCGVDFTESRLSNLLGIRKRQPFQEGFQI LYEDLEGGNIPALLDVEAYEESKEKAEAEATTAVATAATVADATVTRGDTFATQAEEA AALAATDDSESKIVIKPVEKDSKNRSYNVLPDGKNTAYRSWYLAYNYGDPEKGVRSWT LLTTSDVTCGVEQVYWSLPDMMQDPVTFRSTRQVSNYPVVGAELLPVYSKSFFNEQAV YSQQLRAFTSLTHVFNRFPENQILVRPPAPTITTVSENVPALTDHGTLPLRSSIRGVQ RVTVTDARRRTCPYVYKALGVVAPRVLSSRTF'' intron complement(14013..26296) /gene=''E2B'' /note=''precedes DNA polymerase and terminal protein precursor pTP CDSs'' CDS 15515..16099 /gene=''L2'' /product=''core protein precursor pVII'' /translation=''MSILISPSNNTGWGLRAPSKMYGGARQRSTQHPVRVRGHFRAPW GALKGRVRSRTTVDDVIDQVVADARNYTPAAAPASTVDAVIDSVVADARRYARAKSRR RRIARRHRSTPAMRAARALLRRARRTGRRAMLRAARRAASGSSSAGRTRRRAATAAAA AIASMSRPRRGNVYWVRDAATGVRVPVRTRPPRT'' CDS 16144..17181 /gene=''L2'' /product=''core protein V'' /translation=''MSKRKYKEEMLQVIAPEIYGPAAAVKEERKPRKLKRVKKDKKEE EDDGLVEFVREFAPRRRVQWRGRKVKPVLRPGTTVVFTPGERSGSASKRSYDEVYGDE DILEQAVERLGEFAYGKRSRPAPLKEEAVSIPLDHGNPTPSLKPVTLQQVLPSAAPRR GFKREGGEDLYPTMQLMVPKRQKLEDVLEHMKVDPEVQPEVKVRPIKQVAPGLGVQTV DIKIPTEPMETQTEPVKPSTSTMEVQTDPWMPAASTTTTRRRRKYGAASLLMPNYALH PSIIPTPGYRGTRFYRGYTSSRRRKTTTRRRRRSRRSSTATSALVRRVYRSGREPLTL PRARYHPSIAI'' CDS 17204..17437 /gene=''L2'' /product=''core protein precursor pX'' /translation=''MALTCRLRVPITGYRGRKPRRRRLTGNGLRRHHHRRRRAISKRL GGGFLPALIPIIAAAIGAIPGIASVAVQASQRH'' regulatory 17461..17466 /regulatory_class=''polyA_signal_sequence'' /gene=''L2'' CDS 17509..18237 /gene=''L3'' /product=''capsid protein precursor pVI'' /translation=''MEDINFSSLAPRHGTRPFMGTWSDIGNSQLNGGAFNWSSLWSGL KNFGSTLKTYGNKAWNSSTGQALREKLKEQNFQQKVVDGLASGINGVVDLANQAVQKQ INSRLDAVPPAGSVEMPQVEEELPPLDKRGDKRPRPDAEETLLTHTDEPPPYEEAVKL GLPTTRPVAPLATGVLKPSSSSQPATLDLPPPASRPSTVAKPLPPVAVASRAPRGRPQ ANWQSTLNSIVGLGVQSVKRRRCY'' CDS 18329..21154 /gene=''L3'' /note=''capsid protein II'' /product=''hexon'' /translation=''MATPSMLPQWAYMHIAGQDASEYLSPGLVQFARATDTYFSLGNK FRNPTVAPTHDVTTDRSQRLTLRFVPVDREDNTYSYKVRYTLAVGDNRVLDMASTYFD IRGVLDRGPSFKPYSGTAYNSLAPKGAPNTSQWKDSDSKMHTFGVAAMPGVVGKKIEA DGLPIGIDSSSGTDTIIYADKTFQPEPQVGSDSWVDTNGAEEKYGGRALKDTTNMKPC YGSFARPTNKEGGQANIKDSETASTTPNYDIDLAFFDSKNIAANYDPDIVMYTENVEL QTPDTHIVFKPGTSDESSEANLGQQAMPNRPNYIGFRDNFIGLMYYNSTGNMGVLAGQ ASQLNAVVDLQDRNTELSYQLLLDSLGDRTRYFSMWNQAVDSYDPDVRIIENHGVEDE LPNYCFPLNGVGFTDTYQGVKVKTDTAATGTNGTQWDKDDTTVSTANEIHSGNPFAME INIQANLWRNFLYANVALYLPDSYKYTPANITLPTNTNTYDYMNGRVVAPSLVDAYIN IGARWSLDPMDNVNPFNHHRNAGLRYRSMLLGNGRYVPFHIQVPQKFFAIKSLLLLPG SYTYEWNFRKDVNMILQSSLGNDLRTDGASIAFTSINLYATFFPMAHNTASTLEAMLR NDTNDQSFNDYLSAANMLYPIPANATNVPISIPSRNWAAFRGWSFTRLKTRETPSLGS GFDPYFVYSGSIPYLDGTFYLNHTFKKVSITFDSSVSWPGNDRLLTPNEFEIKRTVDG EGYNVAQCNMTKDWFLVQMLAHYNIGYQGFYVPEGYKDRMYSFFRNFQPMSRQVVDEV NYKDYQAVTLAYQHNNSGFVGYLAPTMRQGQPYPANYPYPLIGKSAVASVTQKKFLCD RVMWRIPFSSNFMSMGALTDLGQNMLYANSAHALDMNFEVDPMDESTLLYVVFEVFDV VRVHQPHRGVIEAVYLRTPFSAGNATT'' CDS 21182..21802 /gene=''L3'' /EC_number=''3.4.22.39'' /product=''protease'' /translation=''TACGSGEQELRAILRDLGCGPCFLGTFDKRFPGFMAPHKLACAI VNTAGRETGGEHWLAFAWNPRSHTCYLFDPFGFSDERLKQIYQFEYEGLLRRSALATE DRCITLEKSTQTVQGPRSAACGLFCCMFLHAFVHWPDRPMDKNPTMNLLTGVPNGMLQ SPQVEPTLRRNQEALYRFLNAHSAYFRSHRARIEKATAFDRMNQDM'' regulatory 21834..21839 /regulatory_class=''polyA_signal_sequence'' /gene=''L3'' gene complement(21877..26364) /gene=''E2A'' gene complement(21877..25341) /gene=''E2A-L'' regulatory complement(21877..) /regulatory_class=''polyA_signal_sequence'' /note=''E2A, E2A-L'' CDS complement(21882..23417) /gene=''E2A'' /product=''single-stranded DNA-binding protein'' /translation=''MAGRGGSQSEQRRQERTPERGRGSASRPPNRESPSPPPLPQKRH AYRRVVSDDGQEEEIVVVSENSRSPSTSPPPPLPPKKKPRKTKHVPLQDISQDSEDER QAEEELAAVGFSFPPVRITEKDGKRVFETLDENDPLTSAAATKMTVKNPLSLPIVSAW EKGMEVMTLLMERYRVESDLKSAFQLMPEQGEVYRRICHLYVNEEHRGIPLTFTSNKT LTTMMGRFLQGFVHSHSQIAHKNWECTGCALWLHGCTEAEGKLRCLHGTVMIQKEHTI EMDVASENGQRALKENPDRAKITQNRWGRSVVQLANNDARCCVHDAGCATNQFSSKSC GVFFTEGGKAQQAFRQLEAFMKAMYPGMSSEQAQMMLIPLHCDCNHKPGCVPSMGRQT CKMTPFGMANAEDLDVEGITDATVLASVKHPALMVFQCCNPVYRNSRAQNAGPNCDFK ISAPDLLGALQLTRKLWSDSFPDLPVPKLLIPEFKWLPKYQFRNVSLPAGHAESRQNP FDF'' intron complement(23427..24079) /note=''E2A, E2A-L; precedes single-stranded DNA-binding protein CDS'' CDS 23443..25842 /gene=''L4'' /product=''hexon assembly protein 100K'' /translation=''METQPSSPTSPSAPAADENQQQQNESLTAPPPSPTSDAAAAPDM QEMEESIEIDLGYVTPAEHEEELAARFSAPEENHQEQPEQEAESEQQQAGLEHGDYLS GAEDVLIKHLARQSIIVKDALLDRAEVPLSVAELSRAYERNLFSPRVPPKRQPNGTCE PNPRLNFYPVFAVPEALATYHLFFKNQRIPVSCRANRTRADALLNLGPGARLPDITSL EEVPKIFEGLGSDETRAANALQGSGEEHEHHSALVELEGDNARLAVLKRTVELTHFAY PALNLPPKVMSAVMDQVLIKRASPLSEEEMQDPESSDEGKPVVSDEQLARWLGASSTP QSLEERRKLMMAVVLVTVELECLRRFFADAETLRKVEENLHYLFRHGFVRQACKISNV ELTNLVSYMGILHENRLGQNVLHTTLRGEARRDYIRDCVYLYLCHTWQTGMGVWQQCL EEQNLKELCKLLQKNLKALWTGFDERTTASDLADLIFPERLRLTLRNGLPDFMSQSML QNFRSFILERSGILPATCSALPSDFVPLTFRECPPPLWSHCYLLRLANYLAYHSDVIE DVSSEGLLECHCRCNLCTPHRSLACNPQLLSETQIIGTFELQGPGEGKGGLKLTPGLW TSAYLRKFVPEDYHPFEIRFYEDQSQPPKAELSACVITQGAILAQLQAIQKSRQEFLL KKGHGVYLDPQTGEELNPSFPQDAPRKQQEAESGAAAAAGGFGGRLGEQSGRGDGRLG QHSGRGGQPARQSGGGRRGGGGGRGRSSRRQTVVLGGGESKQHGYHLRSGSGSRRPGP Q'' intron complement(24157..26296) /note=''E2A, E2B'' intron complement(24157..25253) /gene=''E2A-L'' regulatory complement(25336..25341) /regulatory_class=''TATA_box'' /gene=''E2A-L'' CDS join(25556..25886,26056..26399) /gene=''L4'' /product=''protein 33K'' /translation=''MPRGSSKKLKVELPLPPEDLEEDWESSQAEEMEDWDSTQAEEDS LQDSLEEEDEVEEEAEEEAAAARPSSSAEEKASSTDTISAPGRGRGGRAHSRWDETGR FPNPTTQTAPTTVSKKRQKPSSSSRKPAAAAAARKSTAAAGGLRIAANEPAQTRELRN RIFPTLYAIFQQSRGQEQELKVKNRSLRSLTRSCLYHKSEDQLQRTLEDAEALFNKYC ALTLKE'' CDS 25556..26125 /gene=''L4'' /product=''encapsidation protein 22K'' /translation=''MPRGSSKKLKVELPLPPEDLEEDWESSQAEEMEDWDSTQAEEDS LQDSLEEEDEVEEEAEEEAAAARPSSSAEEKASSTDTISAPGRGRGGRAHSRWDETGR FPNPTTQTGKKERQGYKSWRGHKNAIVSCLQACGGNISFTRRYLLFHRGVNFPRNILH YYRHLHSPYYCFQEEAETQQQQQKTSGSS'' intron 25887..26055 /gene=''L4'' regulatory complement(26388..26393) /regulatory_class=''TATA_box'' CDS 26471..27154 /gene=''L4'' /product=''capsid protein precursor pVIII'' /translation=''MSKEIPTPYMWSYQPQMGLAAGAAQDYSTRMNWLSAGPAMISRV NDIRAHRNQILLEQSAITATPRHHLNPRNWPAALVYQEIPQPTTVLLPRDAQAEVQLT NSGVQLAGGAALCRHRPAQGIKRLVIRGRGTQLNDEVVSSSLGLRPDGVFQLAGSGRS SFTPRQAVLTLESSSSQPRSGGIGTLQFVEEFTPSVYFNPFSGSPGHYPDEFIPNFDA ISESVDGYD'' gene 26836..32075 /gene=''E3'' regulatory 26836..26842 /regulatory_class=''TATA_box'' /gene=''E3'' intron 26888..31546 /gene=''E3'' /note=''precedes control protein E3 14.7K CDS'' intron 26888..31098 /gene=''E3'' /note=''precedes membrane protein E3 RID-beta CDS'' intron 26888..30849 /gene=''E3'' /note=''precedes membrane protein E3 RID-alpha CDS''
intron 26888..29956 /gene=''E3'' /note=''precedes membrane glycoprotein E3 CR1-gamma CDS'' intron 26888..28596 /gene=''E3'' /note=''precedes membrane glycoprotein E3 CR1-beta CDS'' intron 26888..28011 /gene=''E3'' /note=''precedes membrane glycoprotein E3 gp19K CDS'' intron 26888..27370 /gene=''E3'' /note=''precedes membrane glycoprotein E3 CR1-alpha CDS'' CDS 27155..27475 /gene=''E3'' /product=''control protein E3 12.5K'' /translation=''MSHGGAADLARLRHLDHCRRFRCFARDLAEFAYFELPEEHPQGP AHGVRIVVEGGLDSHLLRIFSQRPILVEREQGQTLLTLYCICNHPGLHESLCCLLCTE YNKS'' CDS 27429..28055 /gene=''E3'' /product=''membrane glycoprotein E3 CR1-alpha'' /translation=''MKVFVVCCVLSIIKAEISDYSGLDCGVPAINRSLFFTGNETELQ LQCKPHKKYLTWLFQGSPIAMVNHCDNDGVLLSGPANLTFSTRRSKLQLFQPFLPGTY QCVSGPCHHTFHLIPNTTAPLPATNNQTTHQRHRRDLSESNTTTHTGGELRGRPTSGI YYGPWEVVGLIALGLVAGGLLALCYLYLPCCSYLVVLCCWFKKWGRSP'' regulatory 27464..27469 /regulatory_class=''polyA_signal_sequence'' /gene=''L4'' CDS 28037..28570 /gene=''E3'' /product=''membrane glycoprotein E3 gp19K'' /translation=''MGKITLVSCGVLVAVVLSIVGLGGAAVVKEEKADPCLHFNPDKC QLSFQPDGNRCTVLIKCGWECENVRIEYNNKTRNNTLASVWQPGDPEWYTVSVPGADG SPRTVNNTFIFAHMCDTVMWMSKQYDMWPPTKENIVVFSIAYSLCTALITAIVCLSIH MLIAIRPRNNAEKEKQP'' CDS 28600..29305 /gene=''E3'' /product=''membrane glycoprotein E3 CR1-beta'' /translation=''MASVTALTIFLGLVGTSSTFQHINKTVYAGSNSVLPGHQSHQKV SWYWYDKSNTPVTLCKGHQTPINRSGIFFKCNHNNITLLSITKHYSGTYYGTNFNIKQ DTYYSVTVLDPTTPRTTTKPTTTKRHTKPKTTKKTTVKTTTTRTTTTTEATTSTTLAA TTHTHTELTLQTTNDLIALLQKGDNSTTSNEEIPRSMIGIIVAVVVCMLIIALCMVYY AFCYRKHRLNDKLEHL'' misc_feature 29636..29946 /note=''residual non-functional 3'-region of membrane glycoprotein E3 CR1-gamma CDS that is intact in other members of this species; lacks splice acceptor and 5'-region'' CDS 29961..30857 /gene=''E3'' /product=''membrane glycoprotein E3 CR1-delta'' /translation=''MKAVSALVFCSLIGIVFSAGFLKNLTIYEGENATLVGISGQNVS WLKYHLDGWKDICDWNVTVYTCNGVNLTITNATQDQNGRFKGQSFTRNNGYESHNMFI YDVTVIRNETATTTQMPTTHSSTTTTMQTTQTTTFYTSTQHMTTTTAAKPSSAAPQPQ ALALIAAQPSTTTRTNEQTTDFLSTVESHTTATSSAFSSTANLSSLSSTPISPATTTP SPAPLPTPLKQTEDSGMQWQITLLIVIGLVILAVLLYYIFCRRIPNAHRKPVYKPIVD GQPEPLQVEGGLRNLLFSFTVW'' CDS 30865..31140 /gene=''E3'' /product=''membrane protein E3 RID-alpha'' /translation=''MIPRQFLITILICLLQVCATLALVANASPDCIGPFASYVLFAFV TCICCCSIVCLLITFFQFIDWIFVRIAYLRHHPQYRDQRVARLLRLL'' CDS 31146..31577 /gene=''E3'' /codon_start=1 /product=''membrane protein E3 RID-beta'' /translation=''MRALLLLALLLLVLPRPVNPRSPTQSPEEVRKCKFQEPWKFLKC YRQKSDMHPSWIMIIGIVNILACTLISFVIYPCFDFGWNSPEALYLPPEPDTPPQQPQ AHALPPPQPRPQYMPILDYEAEPQRPMLPAISYFNLTGGDD'' CDS 31570..31977 /gene=''E3'' /note=''12.5K family'' /product=''control protein E3 14.7K'' /translation=''MTDPLANNNVNDLLLDMDGRASEQRLAQLRIRQQQERAVKELQD GIAIHQCKKGIFCLVKQAKISYEVTQTDHRLSYELLQQRQKFTCLVGVNPIVITQQSG DTKGCIHCSCDSPDCVHTLIKTLCGLRDLLPMN'' regulatory 32001..32006 /regulatory_class=''polyA_signal_sequence'' /gene=''E3'' regulatory 32070..32075 /regulatory_class=''polyA_signal_sequence'' /gene=''E3'' CDS 32254..33531 /gene=''L5'' /note=''capsid protein IV'' /product=''fiber'' /translation=''MSKKRVRVDDDFDPVYPYDADNAPTVPFINPPFVSSDGFQEKPL GVLSLRLADPVTTKNGEITLKLGEGVDLDSSGKLISNTATKAAAPLSFSNNTISLNMD HPFYTKDGKLSLQVSPPLNILRTSILNTLALGFGSGLGLRGSALAVQLVSPLTFDTDG NIKLTLDRGLHVTTGDAIESNISWAKGLKFEDGAIATNIGNGLEFGSSSTETGVDDAY PIQVKLGSGLSFDSTGAIMAGNKEDDKLTLWTTPDPSPNCQILAENDAKLTLCLTKCG SQILATVSVLVVGSGNLNPITGTVSSAQVFLRFDANGVLLTEHSTLKKYWGYRQGDSI DGTPYTNAVGFMPNLKAYPKSQSSTTKNNIVGQVYMNGDVSKPMLLTITLNGTDDSNS TYSMSFSYTWTNGSYVGATFGANSYTFSYIAQE'' regulatory 33599..33604 /regulatory_class=''polyA_signal_sequence'' /gene=''L5'' gene complement(33620..36319) /gene=''E4'' regulatory complement(33620..33625) /regulatory_class=''polyA_signal_sequence'' /gene=''E4'' CDS complement(join(33638..33889,34621..34791)) /gene=''E4'' /product=''control protein E4orf6/7'' /translation=''MSESNCIMTRSRARSAASRHHPYRPAPLPRCEETETRASLVEDH PVLPDCDTLSMHNITVIPTTEDNPQLLSCEVQMRECPEGFISLTDPRLARSETVWNVE TKSMSITNGVQMFKAVRGERVVYSMSWEGGGKITARIL'' CDS complement(33886..34791) /gene=''E4'' /note=''E4orf6; 34K family'' /product=''control protein E4 34K'' /translation=''MSESNCIMTRSRARSAASRHHPYRPAPLPRCEETETRASLVEDH PVLPDCDTLSMHNVSSVRGLPCSAGFAVLQEFPVPWDMVLTPEELRVLKRCMSVCLCC ANIDLFSSQMIHGYERWVLHCHCRDPGSLRCMAGGAVLALWFRRIIRGCMFNQRVMWY REVVNRHMPKEIMYMGSVFWRGRHLIYLRIWYDGHVGSILPAMSFGWSVLNYGLLNNL VVLCCTYCSDLSEIRMRCCARRTRRLMLRAVGIMLRESLDPDPLSSSLTERRRQRLLR GLMRHHRPIPFADYDSHRRSSASSR'' intron complement(33890..34620) /gene=''E4'' CDS complement(34697..35062) /gene=''E4'' /product=''control protein E4orf4'' /translation=''MVLPVLPSPAVTETQQNCIIWLGLAHSTVVDVIRAIRHDGIFIT PEALDLLHGLREWLFYNFNTERSKRRDRRRRSVCSARTRFCYSKYENVRKQLHHDTVA STISRVPPSPVSAGPLTTL'' intron complement(34815..36232) /gene=''E4'' /note=''precedes control protein E4 34K CDS'' CDS complement(35072..35425) /gene=''E4'' /product=''control protein E4orf3'' /translation=''MRVCLRMPVEGALRELFIMAGLDLPQELVRIIQGWKNENYLGMV QECNMMIEELENPPAFAIVLFLDVRVEALLEATVEHLENRITFDLAVIFHQHSGGERC HLRDLHFEVLRDRLD'' intron complement(35136..36232) /gene=''E4'' /note=''precedes control protein E4orf4 CDS'' CDS complement(35422..35811) /gene=''E4'' /product=''control protein E4orf2'' /translation=''MLERTACIYSIVVPEALNVHLEDFSFVDFLKNCLGDFLSSYLED ITGSSQHAYSSLAFGNAHWGGLRFICTVACPNLIPGGPMAKNFGEDMKEYLQLLLREE LRDRGREFDIPLVNLLQVNQEQNILEL'' intron complement(35455..36232) /gene=''E4'' /note=''precedes control protein E4orf3 CDS'' intron complement(35827..36232) /gene=''E4'' /note=''precedes control protein E4orf2 CDS'' CDS complement(35851..36225) /gene=''E4'' /note=''genus-specific; DURP family'' /product=''control protein E4orf1'' /translation=''MDAEALYVYLEGSGALLPVQEGSNYILYAPENFVLHPHGIALLD LRLSIVVPYCFLGRFFSLADANVPGVYSSCRIIHAGHRERLSVMVFNHSDNFYEGRAG DPVACLVLERTIYPPVRQASMV'' regulatory complement(36314..36319) /regulatory_class=''TATA_box'' /gene=''E4'' repeat_region 36515..36643 /standard_name=''ITR'' /rpt_type=inverted
Annotation of the ChAd155 wild type sequence (SEQ ID NO: 19) sequence is provided below.
TABLE-US-00002 LOCUS ChAd155 37830 bp DNA linear 10-JUN-2015 DEFINITION Chimp adenovirus 155, complete genome. COMMENT Annotation according to alignment of ChAd155 against the human Adenovirus 2 reference strain NC_001405 Two putative ORFs in the E3 region added manually FEATURES Location/Qualifiers source 1..37830 /organism="Chimpanzee adenovirus 155" /mol_type="genomic DNA" /acronym="ChAd155" repeat_region 1..101 /standard name="ITR" /rpt_type=inverted gene 466..1622 /gene="E1A" TATA_signal 466..471 /gene="E1A" prim_transcript 497..1622 /gene="E1A" CDS join(577..1117,1231..1532) /gene="E1A" /product="E1A_280R" CDS join(577..979,1231..1532) /gene="E1A" /product="E1A_243R" polyA_signal 1600..1605 /gene="E1A" gene 1662..4131 /gene="E1B" TATA_signal 1662..1667 /gene="E1B" prim_transcript 1692..4131 /gene="E1B" CDS 1704..2267 /gene="E1B" /product="E1B_19K" CDS 2009..3532 /gene="E1B" /product="E1B_55K" gene 3571..4131 /gene="IX" TATA_signal 3571..3576 /gene="IX" prim_transcript 3601..4131 /gene="IX" CDS 3628..4092 /gene="IX" /product="IX" polyA_signal 4097..4102 /note="E1B, IX" gene complement(4117..27523) /gene="E2B" prim_transcript complement(4117..27494) /gene="E2B" gene complement(4117..5896) /gene="IVa2" prim_transcript complement(4117..5896) /gene="IVa2" CDS complement(join(4151..5487,5766..5778)) /gene="IVa2" /product="E2B_IVa2" polyA_signal complement(4150..4155) /note="IVa2, E2B" CDS complement(join(5257..8838,14209..14217)) /gene="E2B" /product="E2B_polymerase" gene 6078..34605 /gene="L5" gene 6078..28612 /gene="L4" gene 6078..22658 /gene="L3" gene 6078..18164 /gene="L2" gene 6078..14216 /gene="L1" TATA_signal 6078..6083 /note="L" prim_transcript 6109..34605 /gene="L5" prim_transcript 6109..28612 /gene="L4" prim_transcript 6109..22658 /gene="L3" prim_transcript 6109..18164 /gene="L2" prim_transcript 6109..14216 /gene="L1" CDS join(8038. .8457,9722..9742) /gene="L1" /product="L1_13.6K" CDS complement(join(8637..10640,14209..14217)) /gene="E2B" /product="E2B_pTP" gene 10671..10832 /gene="VAI" misc_RNA 10671..10832 /gene="VAI" /product="VAI" gene 10902..11072 /gene="VAII" misc_RNA 10902..11072 /gene="VAII" /product="VAII" CDS 11093..12352 /gene="L1" /product="L1_52K" CDS 12376..14157 /gene="L1" /product="L1_pIIIa" polyA_signal 14197..14202 /gene="L1" CDS 14254..16035 /gene="L2" /product="L2_penton" CDS 16050..16646 /gene="L2" /product="L2_pVII" CDS 16719..17834 /gene="L2" /product="L2_V" CDS 17859..18104 /gene="L2" /product="L2_pX" polyA_signal 1814..18148 /gene="L2" CDS 18196..18951 /gene="L3" /product="L3_pVI" CDS 19063..21945 /gene="L3" /product="L3_hexon" CDS 21975..22604 /gene="L3" /product="L3_protease" polyA_signal 22630..22635 /gene="L3" gene complement(22632..27523) /gene="E2A" prim_transcript complement(22632..27494) /gene="E2A" gene complement(22632..26357) /gene="E2A-L" prim_transcript complement(22632..26328) /gene="E2A-L" polyA_signal complement(22649..22654) /note="E2A, E2A-L" CDS complement(22715..24367) /gene="E2A" /note="DBP; genus-common; DBP family" /codon_start=1 /product="E2A" CDS 24405..26915 /gene="L4" /product="L4_100k" TATA_signal complement(26352..26357) /gene="E2A-L" CDS join(26602..26941,27147..27529) /gene="L4" /product="L4_33K" CDS 26602..27207 /gene="L4" /product="L4_22K" TATA_signal complement(27518..27523) /note="E2A, E2B; nominal" CDS 27604..28287 /gene="L4" /product="L4_pVIII" gene 27969..32686 /gene="E3B" gene 27969..31611 /gene="E3A" TATA_signal 27969..27974 /note="E3A, E3B" prim_transcript 27998..32686 /gene="E3B" prim_transcript 27998..31611 /gene="E3A" CDS 28288..28605 /gene="E3A" /product="E3 ORF1" polyA_signal 28594..28599 /gene="L4" CDS 29103..29303 /gene="E3A" /product="E3 ORF2" CDS 29300..29797 /gene="E3A" /product="E3 ORF3" CDS 29826..30731 /gene="E3A" /product="E3 ORF4" CDS 30728..31579 /gene="E3A" /product="E3 ORF5" CDS 31283..31579 /gene="E3A" /product="E3 ORF6" polyA_signal 31578..31584 /gene="E3A" CDS 31591..31863 /gene="E3B" /product="E3 ORF7" CDS 31866..32264 /gene="E3B" /product="E3 ORF8" CDS 32257..32643 /gene="E3B" /product="E3 ORF9" polyA_signal 32659..32664 /gene="E3B" gene complement(<32678..32838) /gene="U" CDS complement(<32678..32838) /gene="U" /note="exon encoding C terminus unidentified genus-common" /product="protein U" CDS 32849..34585 /gene="L5" /product="L5_fiber" polyA_signal 34581..34586 /gene="L5" gene complement(34611..37520) /gene="E4" prim_transcript complement(34611..37490) /gene="E4" polyA_signal complement(34625..34630) /gene="E4" CDS complement(join(34794..35069,35781..35954)) /gene="E4" /product="E4 ORF7" CDS complement(35070..35954) /gene="E4" /product="E4 ORF6" CDS complement(35875..36219) /gene="E4" /product="E4 ORF4" CDS complement(36235..36582) /gene="E4" /product="E4 ORF3" CDS complement(36579..36971) /gene="E4" /product="E4 ORF2" CDS complement(37029..37415) /gene="E4" /product="E4 ORF1" TATA_signal complement(37515..37520) /gene="E4"
repeat_region 37740..37830 /standard_name="ITR" /rpt_type=inverted
Transgenes
[0391] Adenoviral vectors may be used to deliver desired RNA or protein sequences, for example heterologous sequences, for in vivo expression. A vector may include any genetic element including naked DNA, a phage, transposon, cosmid, episome, plasmid, or a virus. By "expression cassette" (or "minigene") is meant the combination of a selected heterologous gene (transgene) and the other regulatory elements necessary to drive translation, transcription and/or expression of the gene product in a host cell.
[0392] Typically, an adenoviral vector is designed such that the expression cassette is located in a nucleic acid molecule which contains other adenoviral sequences in the region native to a selected adenoviral gene. The expression cassette may be inserted into an existing gene region to disrupt the function of that region, if desired. Alternatively, the expression cassette may be inserted into the site of a partially or fully deleted adenoviral gene. For example, the expression cassette may be located in the site of a mutation, insertion or deletion which renders non-functional at least one gene of a genomic region selected from the group consisting of E1A, E1B, E2A, E2B, E3 and E4. The term "renders non-functional" means that a sufficient amount of the gene region is removed or otherwise disrupted, so that the gene region is no longer capable of producing functional products of gene expression. If desired, the entire gene region may be removed (and suitably replaced with the expression cassette). Suitably, E1 genes of adenovirus are deleted and replaced with an expression cassette consisting of the promoter of choice, cDNA sequence of the gene of interest and a poly A signal, resulting in a replication defective recombinant virus.
[0393] A transgene sequence may also include a reporter sequence, which upon expression produces a detectable signal. Such reporter sequences include, without limitation, DNA sequences encoding .beta.-lactamase, .beta.-galactosidase (LacZ), alkaline phosphatase, thymidine kinase, green fluorescent protein (GFP), chloramphenicol acetyltransferase (CAT), luciferase, membrane bound proteins including, for example, CD2, CD4, CD8, the influenza hemagglutinin protein, and others well known in the art, to which high affinity antibodies directed thereto exist or can be produced by conventional means, and fusion proteins comprising a membrane bound protein appropriately fused to an antigen tag domain from, among others, hemagglutinin or Myc. These coding sequences, when associated with regulatory elements which drive their expression, provide signals detectable by conventional means, including enzymatic, radiographic, colorimetric, fluorescence or other spectrographic assays, fluorescent activating cell sorting assays and immunological assays, including enzyme linked immunosorbent assay (ELISA), radioimmunoassay (RIA) and immunohistochemistry.
[0394] In addition to the transgene the expression cassette also includes conventional control elements which are operably linked to the transgene in a manner that permits its transcription, translation and/or expression in a cell transfected with the adenoviral vector. As used herein, "operably linked" sequences include both expression control sequences that are contiguous with the gene of interest and expression control sequences that act in trans or at a distance to control the gene of interest.
[0395] Expression control sequences include appropriate transcription initiation, termination, promoter and enhancer sequences; efficient RNA processing signals such as splicing and polyadenylation (poly A) signals including rabbit beta-globin polyA; sequences that stabilize cytoplasmic mRNA; sequences that enhance translation efficiency (e.g., Kozak consensus sequence); sequences that enhance protein stability; and when desired, sequences that enhance secretion of the encoded product. Among other sequences, chimeric introns may be used.
[0396] A "promoter" is a nucleotide sequence that permits binding of RNA polymerase and directs the transcription of a gene. Typically, a promoter is located in the 5' non-coding region of a gene, proximal to the transcriptional start site of the gene. Sequence elements within promoters that function in the initiation of transcription are often characterized by consensus nucleotide sequences. Examples of promoters include, but are not limited to, promoters from bacteria, yeast, plants, viruses, and mammals (including humans). A great number of expression control sequences, including promoters which are internal, native, constitutive, inducible and/or tissue-specific, are known in the art and may be utilized.
[0397] Examples of constitutive promoters include, without limitation, the TBG promoter, the retroviral Rous sarcoma virus LTR promoter (optionally with the enhancer), the cytomegalovirus (CMV) promoter (optionally with the CMV enhancer, see, e.g., Boshart et al, Cell, 41:521-530 (1985)), the CASI promoter (WO2012/115980), the SV40 promoter, the dihydrofolate reductase promoter, the .beta.-actin promoter, the phosphoglycerol kinase (PGK) promoter, and the EF1a promoter (Invitrogen).
[0398] Inducible promoters allow regulation of gene expression and can be regulated by exogenously supplied compounds, environmental factors such as temperature, or the presence of a specific physiological state, e.g., acute phase, a particular differentiation state of the cell, or in replicating cells only. Inducible promoters and inducible systems are available from a variety of commercial sources, including, without limitation, Invitrogen, Clontech and Ariad. Many other systems have been described and can be readily selected by one of skill in the art. For example, inducible promoters include the zinc-inducible sheep metallothionine (MT) promoter and the dexamethasone (Dex)-inducible mouse mammary tumor virus (MMTV) promoter. Other inducible systems include the T7 polymerase promoter system (WO 98/10088); the ecdysone insect promoter (No et al, Proc. Natl. Acad. Sci. USA, 93:3346-3351 (1996)), the tetracycline-repressible system (Gossen et al, Proc. Natl. Acad. Sci. USA, 89:5547-5551 (1992)), the tetracycline-inducible system (Gossen et al, Science, 378:1766-1769 (1995), see also Harvey et al, Curr. Opin. Chem. Biol, 2:512-518 (1998)). Other systems include the FK506 dimer, VP16 or p65 using castradiol, diphenol murislerone, the RU486-inducible system (Wang et al, Nat. Biotech., 15:239-243 (1997) and Wang et al, Gene Ther., 4:432-441 (1997)) and the rapamycin-inducible system (Magari et al, J. Clin. Invest., 100:2865-2872 (1997)). The effectiveness of some inducible promoters increases over time. In such cases one can enhance the effectiveness of such systems by inserting multiple repressors in tandem, e.g., TetR linked to a TetR by an IRES.
[0399] The transgene may be operably linked to a tissue-specific promoter. For instance, if expression in skeletal muscle is desired, a promoter active in muscle should be used. These include the promoters from genes encoding skeletal .beta.-actin, myosin light chain 2A, dystrophin, muscle creatine kinase, as well as synthetic muscle promoters with activities higher than naturally occurring promoters (see Li et al, Nat. Biotech., 17:241-245 (1999)). Examples of promoters that are tissue-specific are known for liver (albumin, Miyatake et al, J. Virol, 71:5124-32 (1997); hepatitis B virus core promoter, Sandig et al, Gene Ther., 3:1002-9 (1996); alpha-fetoprotein (AFP), Arbuthnot et al., Hum. Gene Ther., 7:1503-14 (1996)), bone osteocalcin (Stein et al, Mol. Biol. Rep., 24:185-96 (1997)); bone sialoprotein (Chen et al., J. Bone Miner. Res., 11:654-64 (1996)), lymphocytes (CD2, Hansal et al, J. Immunol, 161:1063-8 (1998); immunoglobulin heavy chain; T cell receptor chain), neuronal such as neuron-specific enolase (NSE) promoter (Andersen et al, Cell. Mol. Neurobiol, 13:503-15 (1993)), neurofilament light-chain gene (Piccioli et al, Proc. Natl. Acad. Sci. USA, 88:5611-5 (1991)), and the neuron-specific vgf gene (Piccioli et al, Neuron, 15:373-84 (1995)), among others.
[0400] In some embodiments, the Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element (WPRE) (Zuffrey et al. (1999) J Virol; 73 (4):2886-9) may be operably linked to the transgene.
Adenoviral Vector Construction
[0401] Adenoviral vectors are generated by the modification of the wild type adenovirus to express heterologous genes and/or delete or inactivate undesirable adenoviral sequences. Adenoviral vectors may also have altered replication competency. For example the vector may be replication defective or have limited replication such that it has a reduced ability to replicate in non-complementing cells, compared to the wild type virus. This may be brought about by mutating the virus e.g. by functionally inactivating or deleting a gene involved in replication, for example E1a, E1b, E2, E3 or E4 .
[0402] The adenoviral vectors in accordance with the present invention may comprise a functionally inactivated or deleted E1. Thus the adenoviral vectors according to the invention may be replication defective due to the absence of the ability to express adenoviral E1a and/or E1b. The recombinant adenoviruses may also bear functional inactivations in other genes (see WO 03/000283) for example, deletions in E3 or E4 genes. The adenovirus delayed early gene E3 may be eliminated from the adenovirus sequence which forms part of the recombinant virus. The function of E3 is not necessary to the production of the recombinant adenovirus particle. Thus, it is unnecessary to replace the function of this gene product in order to package a recombinant adenovirus useful in the invention. In one particular embodiment the recombinant adenoviruses have functionally deleted E1 and E3 genes. The construction of such vectors is described in Roy et al., Human Gene Therapy 15:519-530, 2004.
[0403] Recombinant adenoviruses may also be constructed having a functional deletion of the E4 gene. In a particular embodiment, the recombinant adenoviruses have functionally deleted E1 and E4 genes as described in Colloca et al. (2012) Sci. Transl. Med. 4:1-9; Roy et al. (2004) Virol. 324: 361-372. In some embodiments it may be desirable to retain the E4 ORF6 function. In one embodiment, the native E4 ORF6 region may be replaced by a heterologous E4 ORF6, such as from Ad5. Thus, in one particular embodiment, the adenoviral vector may be functionally deleted in E1 and have the E4 ORF6 region from Ad5.
[0404] Adenovirus vectors according to the invention may also contain a functional deletion in the delayed early gene E2a. Deletions may also be made in any of the late genes L1 through to L5 of the adenovirus genome. Similarly deletions in the intermediate genes IX and IVa may be useful.
[0405] Other deletions may be made in the other structural or non-structural adenovirus genes. The above deletions may be used individually, e.g. an adenovirus sequence for use in the present invention may contain deletions of E1 only. Alternatively, deletions of entire genes or portions thereof effective to destroy their biological activity may be used in any combination. For example in one exemplary vector, the adenovirus sequences may have deletions of the E1 genes and the E4 gene, or of the E1, E2a and E3 genes, or of the E1 and E3 genes (such as functional deletions in E1a and E1b, and a deletion of at least part of E3), or of the E1, E2a and E4 genes, with or without deletion of E3 and so on. Such deletions may be partial or full deletions of these genes and may be used in combination with other mutations, such as temperature sensitive mutations to achieve a desired result.
[0406] These vectors are generated using techniques known to those of skill in the art. Such techniques include conventional cloning techniques of cDNA such as those described in texts, use of overlapping oligonucleotide sequences of the adenovirus genomes, polymerase chain reaction, and any suitable method which provides the desired nucleotide sequence. Particularly suitable methods include standard homologous recombination methods such as those provided in Colloca et al. (2012) Sci. Transl. Med. 4:1-9; Roy et al. (2004) Virol. 324: 361-372; Roy et al. (2010) J. of Gene Med. 13:17-25; and WO2010/085984 or recombineering methods as described in Warming et al. Nuc. Acids Res. (2005) 33:e36.
[0407] Suitably, an adenovirus sequence for use in the present invention will contain functional inactivation (such as deletion) of at least the E1 and E4 genes, optionally with E3 functional inactivation (such as deletion), in conjunction with Ad5E4orf6 gene substitution.
[0408] In one embodiment the adenovirus comprises functional inactivation (such as deletion) of the E1 and E4 genes, with incorporation of E4orf6 from Ad5. In such embodiments adenovirus is suitably derived from ChAd155, ChAd3 or ChAd63, particularly ChAd3.
[0409] In a second embodiment the adenovirus comprises functional inactivation (such as deletion) of the E1, E3 and E4 genes, with incorporation of E4orf6 from Ad5. In such embodiments adenovirus is suitably derived from ChAd155, ChAd3 or ChAd63, particularly ChAd63.
Adenoviral Vector Production
[0410] The adenoviral vectors can be produced using any suitable cell line in which the virus is capable of replication. In particular, complementing cell lines which provide the factors missing from the viral vector that result in its impaired replication characteristics (such as E1) can be used. Without limitation, such a cell line may be HeLa (ATCC Accession No. CCL 2), A549 (ATCC Accession No. CCL 185), HEK 293, KB (CCL 17), Detroit (e.g., Detroit 510, CCL 72) and WI-38 (CCL 75) cells, among others. These cell lines are all available from the American Type Culture Collection, 10801 University Boulevard, Manassas, Va. 20110-2209. Other suitable parent cell lines may be obtained from other sources, such as PER.C6.TM. cells, as represented by the cells deposited under ECACC no. 96022940 at the European Collection of Animal Cell Cultures (ECACC) at the Centre for Applied Microbiology and Research (CAMR, UK) or Her 96 cells (Crucell).
[0411] A particularly suitable complementation cell line is the Procell92 cell line. The Procell92 cell line is based on HEK 293 cells which express adenoviral E1 genes, transfected with the Tet repressor under control of the human phosphoglycerate kinase-1 (PGK) promoter, and the G418-resistance gene (Vitelli et al. PLOS One (2013) 8 (e55435):1-9). Procell92.S is adapted for growth in suspension conditions and is also useful for producing adenoviral vectors expressing toxic proteins (www.okairos.com/e/inners.php?m=00084, last accessed 13 Apr. 2015).
Adenoviral Delivery Methods and Dosage
[0412] The adenoviral vectors may be as administered in immunogenic compositions. An immunogenic composition as described herein is a composition comprising one or more recombinant vectors capable of inducing an immune response, for example a humoral (e.g., antibody) and/or cell-mediated (e.g., a cytotoxic T cell) response, against the transgene product delivered by the vector following delivery to a mammal, suitably a human. A recombinant adenovirus may comprise (suitably in any of its gene deletions) a gene encoding the desired immunogen and may therefore be used in a vaccine.
[0413] Such vaccine or other immunogenic compositions may be formulated in a suitable delivery vehicle. Generally, doses for the immunogenic compositions are in the range defined below under `Delivery Methods and Dosage`.
[0414] Optionally, a vaccine or immunogenic composition of the invention may be formulated to contain other components, including, e.g., adjuvants, stabilizers, pH adjusters, preservatives and the like. An adjuvant can be administered with a priming DNA vaccine encoding an antigen to enhance the antigen-specific immune response compared with the immune response generated upon priming with a DNA vaccine encoding the antigen only.
[0415] The adenoviral vector may be prepared for administration by being suspended or dissolved in a pharmaceutically or physiologically acceptable carrier such as isotonic saline; isotonic salts solution or other formulations that will be apparent to those skilled in the art. The appropriate carrier will be evident to those skilled in the art and will depend in large part upon the route of administration. The compositions described herein may be administered to a mammal in a sustained release formulation using a biodegradable biocompatible polymer, or by on-site delivery using micelles, gels and liposomes.
[0416] In some embodiments, the recombinant adenovirus of the invention is administered to a subject by intramuscular injection, intravaginal injection, intravenous injection, intraperitoneal injection, subcutaneous injection, epicutaneous administration, intradermal administration, nasal administration or oral administration. Delivery to the lung may also be desirable. Intramuscular delivery may be a typical route, for reasons of simplicity and convenience.
[0417] If the therapeutic regimen involves co-administration of one or more adenoviral vectors and a further component, each formulated in different compositions, they are favourably administered co-locationally at or near the same site. For example, the components can be administered (e.g. via an administration route selected from intramuscular, transdermal, intradermal, sub-cutaneous) to the same side or extremity ("co-lateral" administration) or to opposite sides or extremities ("contra-lateral" administration).
[0418] Dosages of the viral vector will depend primarily on factors such as the condition being treated, the age, weight and health of the patient, and may thus vary among patients. For example, a therapeutically effective adult human or veterinary dosage of the viral vector generally contains 1.times.10.sup.5 to 1.times.10.sup.15 viral particles, such as from 1.times.10.sup.8 to 1.times.10.sup.12 (e.g., 1.times.10.sup.8, 5.times.10.sup.8, 1.times.10.sup.9, 5.times.10.sup.9, 1.times.10.sup.10, 2.5.times.10.sup.10, 5.times.10.sup.10, 1.times.10.sup.11 5.times.10.sup.11, 1.times.10.sup.12 particles). Alternatively, a viral vector can be administered at a dose that is typically from 1.times.10.sup.5 to 1.times.10.sup.10 plaque forming units (PFU), such as 1.times.10.sup.5 PFU, 5.times.10.sup.5 PFU, 1.times.10.sup.6 PFU, 5.times.10.sup.6 PFU, 1.times.10.sup.7 PFU, 5.times.10.sup.7 PFU, 1.times.10.sup.8 PFU, 5.times.10.sup.8 PFU, 1.times.10.sup.9 PFU, 5.times.10.sup.9 PFU, or 1.times.10.sup.10 PFU. Dosages will vary depending upon the size of the animal and the route of administration. For example, a suitable human or veterinary dosage (for about an 80 kg animal) for intramuscular injection is in the range of about 1.times.10.sup.9 to about 5.times.10.sup.12 particles per mL, for a single site. Optionally, multiple sites of administration may be used. In another example, a suitable human or veterinary dosage may be in the range of about 1.times.10.sup.11 to about 1.times.10.sup.15 particles for an oral formulation.
[0419] The adenoviral vector can be quantified by Quantitative PCR Analysis (Q-PCR), for example with primers and probe designed on CMV promoter region using as standard curve serial dilution of plasmid DNA containing the vector genome with expression cassette including HCMV promoter. The copy number in the test sample is determined by the parallel line analysis method. Alternative methods for vector particle quantification can be analytical HPLC or spectrophotometric method based on A.sub.260 nm.
[0420] Generally a human dose will be in a volume of between 0.5 ml and 2 ml. Thus the composition described herein can be formulated in a volume of, for example 0.5, 1.0, 1.5 or 2.0 ml human dose per individual or combined immunogenic components. A volume of 400-600 ul, such as around 500 ul is typically used, in particular for administration by the intramuscular route.
[0421] One of skill in the art may adjust these doses, depending on the route of administration and the therapeutic or vaccine application for which the recombinant vector is employed. The levels of expression of the transgene, or for an adjuvant, the level of circulating antibody, can be monitored to determine the frequency of dosage administration.
[0422] The therapeutic levels of, or level of immune response against, the protein encoded by the selected transgene can be monitored to determine the need, if any, for boosters. Following an assessment of CD8+T cell response, or optionally, antibody titers, in the serum, optional booster immunizations may be desired. Optionally, the adenoviral vector may be delivered in a single administration or in various combination regimens, e.g., in combination with a regimen or course of treatment involving other active ingredients or in a prime-boost regimen.
M72 Transgene
[0423] A further aspect of the present invention relates to a novel polynucleotide encoding an
[0424] M72 antigen which has been optimised for use in the present invention but will also have utility in other contexts. Consequently, the invention also provides a polynucleotide comprising SEQ ID No: 8 or a degenerate variant thereof having at least 95% identity to SEQ ID No: 8 (such as at least 98% identity, suitably at least 99% identity, in particular at least 99.5% identity and especially 100% identity). Also provided is a polynucleotide consisting of SEQ ID No: 8 or a degenerate variant thereof having at least 95% identity to SEQ ID No: 8 (such as at least 98% identity, in suitably at least 99% identity, particular at least 99.5% identity and especially 100% identity). By the term degenerate variant is meant a variant of the polynucleotide which encodes the same polypeptide.
[0425] The optimised polynucleotide helps ensure the benefits of the present invention are fully achieved through efficient transgene expression in human cells.
Adenoviral Constructs
[0426] A further aspect of the present invention relates to novel adenoviral constructs of use in the present invention but also having utility in other contexts. Consequently, the invention also provides a non-human simian adenovirus comprising a transgene encoding a Rv1196 or Rv0125 related antigen. Suitably the non-human simian adenovirus comprises a penton of SEQ ID No: 10, a hexon of SEQ ID No: 11 or a fibre of SEQ ID No: 12, in particular a penton of SEQ ID No: 10, a hexon of SEQ ID No: 11 and a fibre of SEQ ID No: 12. Alternatively, the non-human simian adenovirus comprises a penton of SEQ ID No: 15, a hexon of SEQ ID No: 16 or a fibre of SEQ ID No: 17, in particular a penton of SEQ ID No: 15, a hexon of SEQ ID No: 16 and a fibre of SEQ ID No: 17. Also, the non-human simian adenovirus may comprise a penton of SEQ ID No: 20, a hexon of SEQ ID No: 21 or a fibre of SEQ ID No: 22, in particular a penton of SEQ ID No: 21, a hexon of SEQ ID No: 22 and a fibre of SEQ ID No: 23.
[0427] The transgene encoding a Rv1196 related antigen may be a sequence encoding a polypeptide comprising, such as consisting of, a sequence having at least 90% identity to SEQ ID No: 1, especially at least 95%, for example at least 98%, such as at least 99% to SEQ ID No: 1, such as SEQ ID No: 1.
[0428] The transgene encoding a Rv0125 related antigen may be a sequence encoding a polypeptide comprising, such as consisting of, a sequence having at least 90% identity to SEQ ID No: 3, especially at least 95%, for example at least 98%, such as at least 99% to SEQ ID No: 3, such as SEQ ID No: 3.
[0429] Suitably the transgene will encode an antigen comprising (such as consisting of) a sequence having at least 90% identity to SEQ ID No. 6. Alternatively, the transgene will encode an antigen comprising (such as consisting of) a fragment of SEQ ID No: 6 which is at least 450 amino acids in length. In some embodiments the transgene will encode an antigen comprising (such as consisting of) amino acids 2-723 of SEQ ID No. 6. Suitably the transgene comprises SEQ ID No: 8 or a degenerate variant thereof having at least 95% identity to SEQ ID No: 8. In some embodiments the transgene comprises SEQ ID No: 8.
[0430] Suitably the adenovirus is replication deficient. For example, the adenovirus comprises functional inactivation (such as deletion) of the E1 gene. The adenovirus may comprise functional inactivation (such as deletion) of the E4 gene. The adenovirus may also comprise functional inactivation (such as deletion) of the E3 gene. The adenovirus may also comprise an Ad5E4orf6 gene substitution.
[0431] Exemplary adenovirus constructs according to the invention are those having the polynucleotide sequence of SEQ ID No: 13 or 18.
[0432] An additional aspect of the invention is a polynucleotide sequence comprising SEQ ID No: 13 or 18, such as a polynucleotide sequence consisting of SEQ ID No: 13 or 18.
Immunisation Regimes, Target Populations and Modes of Administration
[0433] In one embodiment the subject receives a single dose of the polypeptide antigen and a single dose of the associated adenovirus. In other embodiments the subject receives two doses of the polypeptide antigen and a single dose of the associated adenovirus (the additional dose of polypeptide antigen may be given prior to initiation of the standard protein/adeno or adeno/protein methods or following completion of the standard methods). In other embodiments the subject receives one dose of the polypeptide antigen and two doses of associated adenovirus (the additional dose of the associated adenovirus may be given prior to initiation of the standard protein/adeno or adeno/protein methods or following completion of the standard methods). When two doses of adenovirus encoding antigen are provided, they may or may not make use of the same adenovirus strain and insert.
[0434] When the subject receives two doses of polypeptide antigen, suitably the additional dose is one week to three months, in particular two weeks to two months, typically three weeks to six weeks, such as three weeks to five weeks, for example around four weeks prior to initiation or following completion of the standard method.
[0435] When the subject receives two doses of associated adenovirus, suitably the additional dose is one week to three months, in particular two weeks to two months, typically three weeks to six weeks, such as three weeks to five weeks, for example around four weeks prior to initiation or following completion of the standard method.
[0436] The subject to be treated using the method of the invention may be of any age. In one aspect of the invention, the subject is human.
[0437] In one embodiment the subject is an adult human (typically aged 18-60).
[0438] The polypeptide and adenovirus compositions may be administered via various suitable routes, including parenteral, such as intramuscular or subcutaneous administration.
[0439] In one particular embodiment, the one or more of the compositions is administered intradermally. The term intradermally as used herein is intended to refer to the application of antigens into the dermis and/or epidermis of human skin. Intradermal application of an immunogenic composition may be performed by using any cutaneous method known to the skilled person including, but not limited to, delivery using a short needle device (a device comprising a microneedle that is between about 0.2 and about 0.6 mm in length) or delivery using a skin patch. Suitable devices for use with the cutaneous vaccines described herein include short needle devices such as those described in U.S. Pat. Nos. 4,886,499, 5,190,521, 5,328,483, 5,527,288, 4,270,537, 5,015,235, 5,141,496, 5,417,662 and EP1092444. Cutaneous vaccines may also be administered by devices which limit the effective penetration length of a needle into the skin, such as those described in WO99/34850. Also suitable are jet injection devices which deliver liquid vaccines to the dermis via a liquid jet injector or via a needle. Also suitable are ballistic powder/particle delivery devices which use compressed gas to accelerate vaccine in powder form through the outer layers of the skin to the dermis. Skin patches will generally comprise a backing plate which includes a solid substrate. Patches deliver the antigen and adjuvant used in the invention to the dermis or epidermis. In particular embodiment, the patches useful in the present invention comprise a plurality of microprojections. The microprojections may be of any shape suitable for piercing the stratum corneum, epidermis and/or dermis and delivery and antigen and adjuvant to the epidermis or dermis. In a particular embodiment, microprojections are biodegradable and comprise a biodegradable polymer.
[0440] In an alternative approach, the polypeptide may be administered intramuscularly and the adenovirus administered intranasally or via aerosol to the lungs.
[0441] Suitably, both compositions are administered intramuscularly.
[0442] Immunogenic compositions used in the invention may be made by admixing the antigen(s) and the adjuvant. The antigen(s) may be provided in a lyophilized form or in a liquid formulation. A kit may be provided comprising a first container comprising the antigen and a second container comprising the adjuvant.
[0443] Suitably, the compositions according to the present invention have a human dose volume of between 0.05 ml and 1 ml, such as between 0.1 and 0.5 ml, in particular a dose volume of about 0.5 ml, or 0.7 ml. The volume of the second immunogenic composition may be reduced, and e.g. be between 0.05 ml and 0.5 ml, such as between 0.1 and 0.2 ml. The volumes of the compositions used may depend on the delivery route with smaller doses being given by the intradermal route.
[0444] Additional embodiments of the invention include:
[0445] (a) A non-human simian adenovirus comprising a transgene encoding a Rv1196 or Rv0125 related antigen, said adenovirus having at least the penton of SEQ ID No: 20, the hexon of SEQ ID No: 21 or the fibre of SEQ ID No: 22.
[0446] (b) The non-human simian adenovirus according to (a), comprising the penton (SEQ ID No: 20), hexon (SEQ ID No: 21) and fibre (SEQ ID No: 22) protein from ChAd155.
[0447] (c) The non-human simian adenovirus according to either (a) or (b) wherein the encoded antigen comprises a sequence having at least 90% identity to SEQ ID No. 6.
[0448] (d) The non-human simian adenovirus according to any one of (a), (b) or (c) wherein the encoded antigen comprises a fragment of SEQ ID No: 6 which is at least 450 amino acids in length, such as 2-723 of SEQ ID No. 6.
[0449] (e) The non-human simian adenovirus according to any one of (a) to (d), which is a replication deficient adenovirus.
[0450] (f) The non-human simian adenovirus according to any one of (a) to (e), wherein the adenovirus comprises functional inactivation (such as deletion) of the E1 gene.
[0451] (g) The non-human simian adenovirus according to any one of (a) to (f), wherein the adenovirus comprises functional inactivation (such as deletion) of the E4 gene.
[0452] (h) The non-human simian adenovirus according to any one of (a) to (g), wherein the adenovirus comprises functional inactivation (such as deletion) of the E3 gene.
[0453] (i) The non-human simian adenovirus according to any one of (a) to (e), wherein the adenovirus comprises an Ad5E4orf6 gene substitution.
[0454] The teaching of all references in the present application, including patent applications and granted patents, are herein fully incorporated by reference. A composition or method or process defined as "comprising" certain elements is understood to encompass a composition, method or process (respectively) consisting of those elements. The invention will be further described by reference to the following, non-limiting, example:
EXAMPLES
Example 1--Generation of ChAd3 and ChAd63 Vectors Encoding M72 Potein
Making Constructs
[0455] An M72 DNA sequence was optimized by GeneArt.RTM. (Life Technologies Corporation) for human expression (SEQ ID No: 8). According to standard methods, the optimised DNA sequence was synthesized and cloned by GeneWiz.RTM. under control of HCMV promoterTetO system and BGH polyA sequences into the shuttle plasmid PVJ using the EcoRV-NotI restriction sites. This plasmid was cleaved with SpeI and SgfI restriction enzymes and recombined into either ChAd3 (with E1 and E4 deletions) or ChAd63 (with E1, E3 and E4 deletions) vectors by homologous recombination in E. coli BJ5183.
[0456] Briefly, the construction of ChAd3 vectors proceeded through the steps provided below.
[0457] The pChAd3 vector is derived from the wild type chimp adenovirus 3 genome. The wild type chimp adenovirus type 3 was isolated from a healthy young chimpanzee housed at the New Iberia Research Center facility (New Iberia Research Center; The University of Louisiana at Lafayette) using standard procedures. The viral genome was then cloned in a plasmid vector and subsequently modified to carry the following modifications in different regions of ChAd3 viral genome:
[0458] 1) deletion of the E1 region (from bp 460 to bp 3543) of the viral genome;
[0459] 2) deletion of the entire ChAd3 E4 coding region (spanning from nucleotide 34634-37349 of ChAd3 wild type sequence) and substitution with Ad5E4orf6 gene. The deleted region all of the E4 region with the exception of E4 native promoter and polyadenylation signal.
[0460] The construction of ChAd 63 vectors proceeded through the steps provided below.
[0461] The wild type chimp adenovirus type 63 was isolated from a healthy group of Chimpanzee housed by the New Iberia facility using standard procedures. The viral genome was then cloned in a plasmid vector and subsequently modified to carry the following modifications in different regions of ChAd3 viral genome:
[0462] 1) deletion of the E1 region (from bp 456 to bp 3421) of the viral genome;
[0463] 2) deletion of the E3 region (from 27208 bp to 31786 bp) of the viral genome;
[0464] 3) deletion of the entire ChAd63 E4 coding region (spanning from nucleotide 33825 to 36216 of ChAd63 wt sequence) and substitution with Ad5E4orf6 gene. The deleted region contained all E4 region with the exception of E4 native promoter and polyadenylation signal.
Confirmatory Testing
[0465] Rescues and viruses amplification (from passage 1 to passage 4) were generated in procell92.S cell line according to standard procedures and the genetic structure of the viral DNAs was checked at passage 3 (M72-ChAd63) or passage 4 (M72-ChAd3) by two different restriction patterns. Each recombinant virus was purified from 1 litre scale culture through a CsCl gradient method. Purified viruses were titred by quantitative PCR and the infectivity measured by a hexon immunostaining method.
[0466] Good M72 expression was confirmed by Western Blot, after HeLa cell line infection with purified viruses. Genomic stability was evaluated until passage 10 and the DNA sequence of complete expression cassette were confirmed by sequencing.
Example 2--Adenovirus Dose Investigation in Mice
Test Groups
[0467] The aim of this study was to assess and compare the immunogenicity of 2 chimpanzee adenoviruses encoding for the tuberculosis M72 antigen: M72-ChAd3 and M72-ChAd63. The adenoviruses were produced according to Example 1.
[0468] Female 6 week old CB6F1/OlaHsd mice, 12 mice per group, were injected by the intramuscular route with 50 ul at day 0 (ChAd 3 solution: pH 7.4, 10 mM TRIS, 10 mM histidine, 5% sucrose, 75 mM NaCl, 1 mM MgCl.sub.2, 0.02% polysorbate 80, 0.1 mM EDTA, 0.5% (v/v) ethanol; ChAd63 solution: pH 6.6, 10 mM histidine, 7.5% sucrose, 35 mM NaCl, 1 mM MgCl.sub.2, 0.1% polysorbate 80, 0.1 mM EDTA, 0.5% (v/v) ethanol):
TABLE-US-00003 Number of Viral Group Adenovirus Particles 1 M72-ChAd3 10{circumflex over ( )}10 2 M72-ChAd3 10{circumflex over ( )}9 3 M72-ChAd3 10{circumflex over ( )}8 4 M72-ChAd3 10{circumflex over ( )}7 5 M72-ChAd63 10{circumflex over ( )}10 6 M72-ChAd63 10{circumflex over ( )}9 7 M72-ChAd63 10{circumflex over ( )}8 8 M72-ChAd63 10{circumflex over ( )}7
[0469] In order to have sufficient volume, the whole blood of 4 pools of 3 mice for groups was collected at days 7, 14, and 21.
Measurement of Cellular Immune Response-Intracellular Cytokine Staining (ICS)
Leukocyte Isolation from Whole Blood
[0470] At each time point, blood was collected from each mouse and subsequently pooled (4 pools of 3 mice). Blood was collected in tubes containing, RPMI/additives (RPMI 1640, supplemented with glutamine, penicillin/streptomycin, sodium pyruvate, non-essential amino-acids and 2-mercaptoethanol) containing heparin 5000 unit/ml (Heparin Leo). Ten volumes of Lysing buffer were added to the whole blood and tubes were incubated at room temperature (RD for 10 min. After centrifugation (335 g, 10 min at RT), the pellet was harvested in RPMI/additives and filtered (Cell strainer 100 um). Cells were pelleted again (335 g, 10 min at RT) and re-suspended in Complete Medium (RPMI 1640, supplemented with glutamine, penicillin/streptomycin, sodium pyruvate, non-essential amino-acids and 2-mercaptoethanol, and 5% heat inactivated fetal calf serum).
In Vitro Stimulation of Fresh Leukocytes
[0471] Leukocytes were plated in round bottom 96-well plates at approximately 1 million cells per well. Leukocytes were then stimulated for 6 hours (37.degree. C., 5% CO.sub.2) with anti-CD28 (clone 37.51) and anti-CD49d (clone 9C10) at 1 ug/ml, with or without 1 ug/ml of peptides covering the M72 sequence (mixture of 15-mer peptides overlapping by 11 amino acid residues). After a 2 hour stimulation period, BD GolgiPlug.TM. containing brefeldin A diluted in complete medium (final dilution 1/1000) was added for 4 additional hours. Plates were then transferred at 4.degree. C., overnight.
ICS IFNg, IL-2, TNF-a
[0472] Cells were stained and analysed using a 5-colour ICS assay.
[0473] Cells were transferred to V-bottom 96-well plates, centrifuged at 189 g for 5 min at 4.degree. C. after wash with 200 ul Flow Buffer (PBS 1.times., 1% FCS), re-suspended the cells in 50 ul Flow Buffer containing anti-CD16/32 (clone 2.4G2) diluted 1/50, for 10 min at 4.degree. C. Then, 50 ul Flow Buffer containing anti-CD4-V450 (clone RM4-5, diluted 1/50) and anti-CD8-PerCp-Cy5.5 (clone 53-6.7, diluted 1/50) antibodies and LIVE/DEAD.RTM. Pacific Orange (Life Technologies, diluted 1/500) was added for 30 min at 4.degree. C. Cells were centrifuged (189 g for 5 min at 4.degree. C.) and washed with 200 ul Flow Buffer.
[0474] Leukocytes were fixed and permeabilised by adding 200 ul of Cytofix/Cytoperm solution (Becton Dickinson commercial buffer) for 20 min at 4.degree. C. Cells were centrifuged (189 g for 5 min at 4.degree. C.) and washed with 200 ul Perm/Wash buffer (Becton Dickinson commercial buffer diluted 1:10 in distilled water). After an additional centrifugation step, cells were stained in 50 ul Perm/Wash buffer with anti-IL2-FITC (clone JES6-5H4, diluted 1/400), anti-IFNg-APC (clone XMG1.2, diluted 1/200) and anti-TNFa-PE (clone MP6-XT22, diluted 1/700) antibodies, for 1 hour at 4.degree. C. Cells were washed twice with the Perm/Wash buffer re-suspended in 220 ul PBS. Stained cells were analysed by flow cytometry using a LSRII and the FlowJo software.
Results
[0475] As seen in FIGS. 1 and 2 (CD4 T cell response for M72-ChAd3 and M72-ChAd63 constructs respectively) and FIGS. 3 and 4 (CD8 T cell response for M72-ChAd3 and M72-ChAd63 constructs respectively) a dose of 1.times.10.sup.8 viral particles per mouse induced the highest level M72 specific response at timepoint 14PI for both the M72-ChAd3 and M72-ChAd63 constructs.
Example 3--Investigation of the Impact of Adenovirus and Adjuvant Coformulation on Infectivity
[0476] The impact of the co-formulation was evaluated with recombinant eGFP-ChAd3 or eGFP-ChAd63. These ChAd3 and ChAd 63 are controls constructed to express the green fluorescent protein instead of the M72 transgene in the respective ChAdenovirus backbones (E1-E4 deleted ChAd3 and E1-E3-E4 deleted ChAd63). Co-formulation with the AS01E (a liposomal formulation of the TLR4 agonist 3D-MPL and the saponin QS21) was evaluated via an infectivity assay based on HeLa cells.
Materials and Methods
Infectivity Test
[0477] HeLa cells were grown in an exponential growth and seeded for 24 h before infection. HeLa cells were used between passage P45 and P65 (Molbiol; GSK Rix).
DAY 0: Cell Harvest
[0478] The medium was removed from the flask and the cells rinsed carefully with DPBS to remove residual cell medium. 5 ml of Trypsin-EDTA was added onto the cells followed by observing the cells under an inverted microscope until the cell layer detached and dispersed (2 to 4 minutes). The cell suspension was gently pipetted up and down and transferred into the Falcon tube and centrifuged at 1200 rpm for 5 to 10 minutes at room temperature. The cell pellet was re-suspended in an appropriate volume and counted. The cells were seeded in a 96 well-plate at 1.5.times.10.sup.4 cells/well (HeLa cells are expected to be at 3.times.10.sup.4 cells/well at the day of infection
DAY 1: Infection Day
[0479] The HeLa cells were observed and were between 50% to 80% confluent. The entire medium was removed from the cells. The recombinant eGFP-ChAd3 and eGFP-ChAd63 stocks were diluted down to a final titre of 5.times.10.sup.7 vp/ml in 80 ul of complete dulbecco modified eagle medium (DMEM). This volume is for 1 well. 80 ul of each sample was added in each well to infect (this is done in duplicate). A negative control of uninfected HeLa cells with a formulation with buffers alone in complete DMEM was used to measure any negative impact on the adeno-infectivity due to the adjuvant buffer. A positive infection control of HeLa cells infected with eGFP-ChAd3 (same condition of infection) was processed in identical conditions. After 3 hours at 37.degree. C., 5% CO.sub.2, 120 ul of complete DMEM was added and then cultured at 37.degree. C., 5% CO.sub.2 for approximately 24 hours.
Day 2: Harvest and FACS Read Out
[0480] The cell supernatants were harvested in a 96 well-plate. Hela cells were rinsed with 40 .mu.l of trypsin/EDTA and then incubated with an additional 40 .mu.l of the trypsin/EDTA mix. Once the cells were detaching from the plate, each well was then gently flushed to recover all the cells. The 96-well plate was then centrifuged at 1200 rpm for 10 minutes. The supernatants were discarded. The cells were suspended in 200 ul of DPBS and kept at 4.degree. C. until FACS acquisition was done on them (LSRII Beckman Dickinson).
Results
TABLE-US-00004
[0481] Test % GFP % viability Group Description A B Average (average) 1 DMEM control 0 0 0 89.14 2 AS01E 0 0 0 91.20 3 AS01E + 81.9 84.79 83.35 89.01 eGFP-ChAd3 4 AS01E + 70.43 73.28 71.86 88.95 eGFP-ChAd63 5 AS buffer + 86.4 86.93 86.67 88.57 eGFP-ChAd3 6 AS buffer + 69.59 71.38 70.49 87.49 eGFP-ChAd63 7 eGFP-ChAd3 78.78 81.29 80.04 90.96 8 eGFP-ChAd63 51.43 53.46 52.45 90.41
AS buffer pH 8.0, 10 mM PO.sub.4, 5 mM NaCl, 4.7% sorbitol ChAd3 buffer: pH 7.4, 10 mM TRIS, 10 mM histidine, 5% sucrose, 75 mM NaCl, 1 mM MgCl.sub.2, 0.02% polysorbate 80, 0.1 mM EDTA, 0.5% (v/v) ethanol ChAd63 buffer: pH 6.6, 10 mM histidine, 7.5% sucrose, 35 mM NaCl, 1 mM MgCl.sub.2, 0.1% polysorbate 80, 0.1 mM EDTA, 0.5% (v/v) ethanol
[0482] The data are expressed as the percentage of cells expressing GFP correlating with the number of cells infected by the ChAd-GFP virus. For all conditions, cell viability has been recorded. This was done to assess any possible cell toxicity which could mislead the conclusion. Fortunately, this was not the case as the cell viability through all the conditions was acceptable and comparable.
[0483] According to the data, no negative impact of the AS on either ChAd3- or ChAd63-GFP is observed. The ChAd3-GFP vector co-formulated with AS01E kept its infectivity potential, which was comparable to the ChAd3-GFP alone.
[0484] Same observation was made for ChAd63-GFP. In the latter case, an increase of infectivity was even observed which may be due to the buffers used in which the adenovirus was diluted.
Example 4--Investigation of the Impact of Adenovirus and Adjuvant Coformulation on QS21 Quenching
[0485] The aim of the study was to check the detoxification of QS21.
Characterisations
TABLE-US-00005
[0486] Visual H Osmo Group Description aspect (sticks) (mosm/kg) 1 AS01E Opalescent 5.5-6 291 2 eGFP-ChAd3 at Slightly 5-5.5 289 2.10 .times. 10.sup.9 in AS01E Opalescent 3 eGFP-ChAd63 at Slightly 5-5.5 297 2.10 .times. 10.sup.9 in AS01E Opalescent 4 eGFP-ChAd3 at Clear 5-5.5 292 2.10 .times. 10.sup.9 in AS01E buffer 5 eGFP-ChAd63 at Clear 5-5.5 304 2.10 .times. 10.sup.9 in AS01E buffer 6 eGFP-ChAd3 at Clear 7 425 2.10 .times. 10.sup.9 7 eGFP-ChAd63 at Clear 6-6.5 408 2.10 .times. 10.sup.9
[0487] The formulated samples containing the adjuvant or the adjuvant buffer have an osmolality 285 mOsm/kg. The pH was taken on indicator stick.
Washing of Red Blood Cells
[0488] The red blood cells (10 ml) were centrifuged for 10 minutes at 1600 rpm (550 g) at 4.degree. C. and the supernatant removed. The red blood cells were re-suspended gently with a volume of buffer DPBS equivalent to the original volume (.+-.10 ml). The operation was repeated (min 2-3 times) until the supernatant was clear (reddish staining disappeared, but the supernatant never becomes completely translucent) and then eliminated the last supernatant after washing. The pellet was stored at 4.degree. C. for 3 to 4 days maximum if not used directly (and washed again the day it is used) or was diluted around 10 times in buffer if used the same day.
Pre Dilutions of Red Blood Cells
[0489] Different pre dilutions of the red blood cells were performed. The pre dilution for which one a 100% of lysis is reached with an OD value (540 nm) between 1.5 and 2 was selected.
[0490] The following dilutions were prepared in haemolysis tubes:
TABLE-US-00006 Dilution Red blood cells dPBS 1/10 100 ul 900 ul 1/12.5 100 ul 1150 ul 1/15 100 ul 1400 ul
[0491] The red blood cells were centrifuged (10 ml) for 10 minutes at 1600 rpm (550 g) at 4.degree. C. 100 ul of the predilution was removed, mixed with 900 ul of WFI and centrifuged for 5 minutes at 2000 rpm (900 g). The supernatant was transferred in a cuvette for spectroscopy and measured the OD at 540 nm. The dilution chosen gave a OD between 1.5 and 2.
QS21 Standard Curve Preparation
[0492] The standard curve of QS21 was made from a QS21 working solution (at 2 mg/ml) diluted extemporaneously to 20 ug/ml in PO.sub.4/Sorbitol buffer.
[0493] Determination of lytic activity was carried out by a limit test.
[0494] 1. Limit of detection (LOD) was defined as the lowest concentration of QS21 leading to an OD:
[0495] Higher than the base level (OD>0.1)
[0496] Around three times higher than OD's buffer (the "0 ug" QS21)
[0497] In the ascendant part of the curve
[0498] Determined for each test.
[0499] 2. QS21 lytic activity was held to be positive in the adjuvant samples if the OD for the adjuvant sample was greater than the OD.sub.LOD.
[0500] Example QS21 Curve
TABLE-US-00007 Delta O.D. O.D. (O.D. sample - Samples QS21 (ug) Sample Buffer O.D. buffer) *Pass/fail AS01E 45 0.151 0.122 0.029 PASS ChAd3 in 45 0.148 0.122 0.026 PASS AS01E ChAd63 in 45 0.143 0.122 0.021 PASS AS01E *Pass: if Delta (O.D. sample - O.D. buffer) < LOD (O.D.) test of the day *Fail: if Delta (O.D. sample - O.D. buffer) > LOD (O.D.) test of the day
Conclusion
[0501] When formulating eGFP-ChAd3 and eGFP-ChAd63 separately with AS01E, the adjuvant size remains unchanged after formulation. Furthermore, there is no free QS21 after formulation, as seen by the red blood cell lysis test.
Example 5--M72 Dosage Regimes
[0502] Evaluating the application of non-replicative chimp adeno vectors expressing M72 in the context of a tuberculosis vaccine, by assessing both homologous (ChAd/ChAd) and heterologous (Prot/ChAd or ChAd/Prot) prime-boost vaccination strategies as well as in combination with M72/AS01E given as admixed or in co-administration.
Materials and Methods
Animal Model
[0503] Female mouse CB6F1/OlaHsd--6 weeks old--12 mice per group--were injected by intramuscular route with 50 ul at days 0-28 as indicated in table below.
TABLE-US-00008 Admin D 0 Admin D 28 Gr. Antigen Dose Solution Antigen Dose Solution Benchmark 1 M72/AS01E 1 ug AS buffer M72/AS01E 1 ug AS buffer Priming: Adeno/Boost: Protein 2 M72-ChAd3 10{circumflex over ( )}8vp ChAd3 M72/AS01E 1 ug AS buffer buffer 3 M72-ChAd63 10{circumflex over ( )}8vp ChAd63 buffer 4 eGFP-ChAd3 10{circumflex over ( )}8vp ChAd3 buffer Priming: Protein/Boost: Adeno 5 M72/AS01E 1 ug AS buffer M72-ChAd3 10{circumflex over ( )}8vp ChAd3 buffer 6 1 ug AS buffer M72-ChAd63 10{circumflex over ( )}8vp ChAd63 buffer Priming: Adeno + AS/Boost: Adeno + AS 7 M72-ChAd3/ 10{circumflex over ( )}8vp AS buffer M72-ChAd3/ 10{circumflex over ( )}8vp AS buffer AS01E AS01E Priming: Adeno/Boost: Adeno 8 M72-ChAd3 10{circumflex over ( )}8vp ChAd3 M72-ChAd3 10{circumflex over ( )}8vp ChAd3 buffer buffer 9 M72-ChAd3 10{circumflex over ( )}8vp ChAd3 M72-ChAd63 10{circumflex over ( )}8vp ChAd63 buffer buffer 10 M72-ChAd63 10{circumflex over ( )}8vp ChAd63 M72-ChAd3 10{circumflex over ( )}8vp ChAd3 buffer buffer Combo Priming and Boost; Adeno + Protein 11 M72/AS01E 1 ug AS buffer M72/AS01E 1 ug AS buffer M72-ChAd3 10{circumflex over ( )}8vp M72-ChAd3 10{circumflex over ( )}8vp Combo Combo 12 M72/AS01E 1 ug AS buffer M72/AS01E 1 ug AS buffer Co-Ad 10{circumflex over ( )}8vp (adjuvanted Co-Ad 10{circumflex over ( )}8vp (adjuvanted M72-ChAd3 protein) M72-ChAd3 protein) and ChAd3 and ChAd3 buffer buffer (adeno) (adeno) 13 M72/AS01E 1 ug AS buffer M72/AS01E 1 ug AS buffer eGFPChAd3 10{circumflex over ( )}8vp eGFPChAd3 10{circumflex over ( )}8vp Combo Combo
AS buffer: approx pH 8.0, 10 mM PO.sub.4, 5 mM NaCl, 4.7% sorbitol ChAd3 buffer: pH 7.4, 10 mM TRIS, 10 mM histidine, 5% sucrose, 75 mM NaCl, 1 mM MgCl.sub.2, 0.02% polysorbate 80, 0.1 mM EDTA, 0.5% (v/v) ethanol ChAd63 buffer: pH 6.6, 10 mM histidine, 7.5% sucrose, 35 mM NaCl, 1 mM MgCl.sub.2, 0.1% polysorbate 80, 0.1 mM EDTA, 0.5% (v/v) ethanol
TABLE-US-00009 Time point Fluid Test of collection Comment Whole ICS D 14(14PI) Evaluation of peripheral whole Blood blood responses of TB antigen- specific CD4 and CD8 cells, as determined by the simultaneous measurement of IFNg, IL-2, TNF- a, after restimulation with overlapping 15-mer peptide pools Whole ICS D 42(14PII)* Evaluation of peripheral whole Blood/ blood and lung responses of TB Lung antigen-specific CD4 and CD8 cells, as determined by the simultaneous measurement of IFNg, IL-2, TNF-a, IL-17 after restimulation with overlapping 15-mer peptide pools Serum Serology D 41(13PII)** anti-M72 IgTot *Limitations of +/-36 lung collection per day, therefore the study was performed in several replicates **Due to practical constraints, serum samples were taken 1 day prior to whole blood
[0504] In order to have sufficient volume, the whole blood of 4 pools of 3 mice for groups was collected at days 14, and 42. At day 42, the same process was applied for lungs.
[0505] Due to limitations of collection per day, one pool of each group was treated per day--during 4 days in order to have 4 pools per groups. Individual sera were collected at day 41.
[0506] The mice were identified in order to do the link between PI and PII for the both read-outs ICS and serology.
Cellular Immune Response-Intracellular Cytokine Staining (ICS)
Leukocyte Isolation from Whole Blood
[0507] At each time point, blood was collected from each mouse and subsequently pooled (4 pools of 3 mice). Blood was collected in tubes containing, RPMI/additives (RPMI 1640, supplemented with Glutamine, Penicillin/streptomycin, Sodium Pyruvate, non-essential amino-acids and 2-mercaptoethanol) containing heparin (1/10). Ten volumes of Lysing buffer were added to the whole blood and tubes were incubated at room temperature (RD for 10 min. After centrifugation (335 g, 10 min at RT), the pellet was harvested in RPMI/additives and filtered (Cell strainer 100 .mu.m). Cells were pelleted again (335 g, 10 min at RD and re-suspended in Complete Medium (RPMI 1640, supplemented with Glutamine, Penicillin/streptomycin, Sodium Pyruvate, non-essential amino-acids and 2-mercaptoethanol, and 5% Heat inactivated Fetal Calf Serum).
Leukocyte Isolation from Lung
[0508] Lung was collected in tubes containing, RPMI/additives (RPMI 1640, supplemented with Glutamine, Penicillin/streptomycin, Sodium Pyruvate, non-essential amino-acids and 2-mercaptoethanol). The sample was transferred in to a Petri dish and the specimen cut in to small pieces (approximately 5.times.5 mm). The specimens were re-suspended in a gentleMACS C tube (violet) containing 10 ml of pre-warmed complete medium containing Liberase (0.0625 Ul/ml=50 ul)+DNase (25 ug=25 ul). The C tube was attached upside down into the sleeve of the gentle MACS Dissociator and the Program mouse-lung 02 (40 sec) ran. At the end of the program, the C tube was detached and the C tube incubated for 30 min at 37.degree. C. in a shaking incubator. After the incubation the sample was transferred on to a cell strainer (100 um) placed on a 50 ml falcon tube. The cell strainer was rinsed twice with 5 ml Complete Medium (RPMI 1640, supplemented with Glutamine, Penicillin/streptomycin, Sodium Pyruvate, non-essential amino-acids and 2-mercaptoethanol, and 5% Heat inactivated Fetal Calf Serum). Complete Medium was added until 45 ml. The cells were centrifuged (400g for 10 min--4.degree. C.), aspirated the supernatant and then re-suspended cells in 10 ml Complete Medium. This step of washing was repeated. After the second wash, the cells were re-suspended in 3 ml Percoll P30. The Percoll P30 layer containing the cells was placed on top of the Percoll P40/P75 layers gradient. The cells were centrifuged for 20 mins 754 g at RT. The cells considered were at the interphase between P40 and P75 at the end of centrifugation (lower interphase). The P30 layer was aspirated and cells collected at interphase P40-P75. The cells were collected in 15 ml tube and Complete Medium added to a final volume of 15 ml. The cells were centrifuged (900 g for 10 min--4.degree. C.). The supernatant was aspirated and the cells washed with 15 ml Complete Medium twice. The pellet was re-suspended with Complete Medium to a volume final of 250 ul. The cells were counted with Macsquant method including viability with PI.
In Vitro Stimulation of Fresh Leukocytes
[0509] Leukocytes were plated in round bottom 96-well plates at approximately 1 million cells per well. Leukocytes were then stimulated for 6 hours (37.degree. C., 5% CO.sub.2) with anti-CD28 (clone 37.51) and anti-CD49d (clone 9C10) at 1 ug/ml, with or without 1 ug/ml of peptides covering the M72 sequence. After a 2 hour-stimulation, BD GolgiPlug.TM. containing brefeldin A diluted in complete medium (final dilution 1/1000) was added for 4 additional hours. Plates were then transferred at 4.degree. C., overnight.
ICS IFNg, IL-2, TNF-a--at 14PI--WBLO
[0510] Cells were stained and analyzed using a 5-colour ICS assay.
[0511] Cells were transferred to V-bottom 96-well plates, centrifuged at 189 g for 5 min at 4.degree. C. after wash with 200 ul Flow Buffer (PBS 1.times., 1% FCS), re-suspended the cells in 50 ul Flow Buffer containing anti-CD16/32 (clone 2.4G2) diluted 1/50, for 10 min at 4.degree. C. Then, 50 ul Flow Buffer containing anti-CD4-V450 (clone RM4-5, diluted 1/50) and anti-CD8-PerCp-Cy5.5 (clone 53-6.7, diluted 1/50) antibodies and Live&Dead PO (diluted 1/500) was added for 30 min at 4.degree. C. Cells were centrifuged (189 g for 5 min at 4.degree. C.) and washed with 200 ul Flow Buffer.
[0512] Leukocytes were fixed and permeabilized by adding 200 ul of Cytofix/Cytoperm solution (Becton Dickinson commercial buffer) for 20 min at 4.degree. C. Cells were centrifuged (189 g for 5 min at 4.degree. C.) and washed with 200 ul Perm/Wash buffer (Becton Dickinson commercial buffer diluted 1:10 in distilled water). After an additional centrifugation step, cells were stained in 50 ul Perm/Wash buffer with anti-IL2-FITC (clone JES6-5H4, diluted 1/400), anti-IFNg-APC (clone XMG1.2, diluted 1/200) and anti-TNFa-PE (clone MP6-XT22, diluted 1/700) antibodies, for 1 hour at 4.degree. C. Cells were washed twice with the Perm/Wash buffer re-suspended in 220 ul PBS. Stained cells were analyzed by flow cytometry using a LSRII and the FlowJo software.
ICS IFNg, IL-2, TNF-a and IL-17 at 14PII--WBLO & Lung
[0513] The same protocol was used except for the step cytokine; Cells were stained in 50 ul Perm/Wash buffer with anti-IL2-FITC (clone JES6-5H4, diluted 1/400), anti-IFNg-APC (clone XMG1.2, diluted 1/200) and anti-TNFa-PE (clone MP6-XT22, diluted 1/700), anti-IL17 BV786 (clone TC11-18H10, diluted 1/50) antibodies, for 1 hour at 4.degree. C.
Humoral Response--Anti-M72 Ig Tot Serology by Elisa
[0514] 96-well Elisa plates were coated with the recombinant antigen M72 at 0.25 ug/ml in PBS and incubated overnight at 4.degree. C. Sera from vaccinated mice at Post II were diluted at 1/5000 or 1/40000 for repeat, in PBS (0.2%)-BSA and then a 2 fold serial dilution is performed from well 1 to 12 and incubated. Serial dilutions of the standard and control material were used to calculate the anti-M72 antibody standard titres of tested sera and to ensure validity of the test. Plates were washed with PBS 0.1% tween20 buffer after each incubation step. A biotinylated goat antibody specific for mice Ig is then added and the antigen-antibody complex is revealed by incubation with a streptavidin-peroxidase complex and a peroxidase substrate ortho-phenylenediamine dihydrochlorid/H.sub.2O.sub.2. The Optical densities (O.D.) were recorded at 490-620 nm. The anti-M72 antibody titre of each individual mouse serum is determined from the standard curve of the ELISA using a regression model and expressed in ELISA unit (EU)/ml. Geometric Mean Titres (GMT) are then calculated for each group of mice.
Results
T Cell Responses
[0515] A. M72-specific CD4 T & CD8 T Cells Responses
[0516] To evaluate the application of non-replicative chimp adeno vectors expressing M72 in the context of a tuberculosis vaccine, both homologous (ChAd/ChAd) and heterologous (Prot/ChAd or ChAd/Prot) prime-boost vaccination strategies were assessed as well as in combination with M72/AS01E4 given as admixed or in co-administration. The prime/boost and combo strategies were evaluated in a D0-D28 schedule. Whole blood was collected at 14PI and 14PII to assess the systemic induction of M72 specific CD4 and CD8 T cells.
[0517] Across all groups, a specific CD4 T cell response was observed in whole blood with the peak response being below 3% (FIGS. 5 and 6). The highest levels of M72 specific CD4 T cells in whole blood were seen with the heterologous (Prot/ChAd or ChAd/Prot) vaccine strategies (FIG. 5). Priming the mice with M72-ChAd vectors and boosting with M72/AS01E induced higher level of M72 specific CD4 T cells than the opposite and in both cases the M72-ChAd3 was more potent than the M72-ChAd63 (FIG. 5). Prime-boost vaccination with M72-ChAd vectors, either homologous (M72-ChAd3/M72-ChAd3)+/-AS01E, or heterologous (M72-ChAd3/M72-ChAd63 or M72-ChAd63/M72-ChAd3), did not provide an added value in terms of the magnitude of the CD4 T cell response as compared to the M72/AS01E benchmark (FIG. 6). Combining both M72-ChAd3 and M72/AS01E slightly induced higher level of M72 specific CD4 T cells as compared to the M72/AS01E benchmark, but lower than heterologous (Prot/ChAd or ChAd/Prot) prime-boost vaccination. Co-administration induced similar M72 specific CD4 T cells levels than the combinations suggesting that physical proximity is not required (FIG. 6).
[0518] The level of M72 specific CD8 T cell was found to be highly increased in mice receiving a M72-ChAd vector either as a prime, boost or both. When the M72-ChAd vector was included during the priming, the level of M72 specific CD8 T cell did not further increase after boosting except when mice where primed with M72-ChAd63 and boosted with M72-ChAd3 (FIGS. 7 and 8). This also suggests that the M72-ChAd3 is more potent than M72-ChAd63. Boosting the M72/AS01E priming with M72-ChAd3 induced higher levels of CD8 T cells (Median=30%) than with M72-ChAd63 (Median=17%). The addition of M72-ChAd3 to the adjuvanted M72 protein in a combo also highly increased the level of M72 specific CD8 T cells (Median=13%) as compared to the current benchmark (Median=0.2%).
[0519] The same general pattern of response was observed in the lung (FIG. 9) as compared to the whole blood (FIGS. 5 and 6) except for the admixed combination vaccine strategy. In the lungs, combination strategy induced a lower level of response than whole blood, whereby a comparable level of M72 specific CD4 T cells could be observed with the combination vaccine and with heterologous (Prot/ChAd or ChAd/Prot) prime-boost vaccination, all of which showed an increased CD4 T cell level as compared to the current benchmark.
[0520] The general pattern of the M72 specific CD8 T cell response in the lungs (FIG. 10) also reflected what was observed in the whole blood (FIGS. 7 and 8). Very low levels of specific CD8 T cell response was observed when mice received the benchmark M72/AS01E (FIG. 10) and addition of M72-ChAd vector highly improved the level of CD8 T cells. The highest level of M72 specific CD8 T cell was seen when mice where primed with M72/AS01E and boosted with M72-ChAd3 (Median=36%--FIG. 10).
[0521] B. Cytokine Profile of the M72-specific CD4 & CD8 T Cells Responses
[0522] In groups that were primed with M72/AS01E, the M72-specific CD4+T cell response mostly included double (IL-2/TNFa) secreting cells in the whole blood at 14PI (FIGS. 11 and 12). In contrast, priming with a M72-ChAd vector induced a polyfunctional M72 specific CD4 T cell response with a majority of triple positive (IL2/IFNg/TNFa), and to a lower extend, double (IFNg/TNFa) and single (IFNg only) producing CD4 T cells (FIGS. 11 and 12). Combining both the protein and the ChAd vector induced low levels of triple (IL2/IFNg/TNFa), and double (IFNg/TNFa) producing CD4 T cells (FIGS. 11 and 12).
[0523] A similar CD4+T cell cytokine expression profiles was observed at 14PII in whole blood (FIGS. 13 and 14) and in the lungs (FIGS. 15 and 16) across all groups where vaccination strategy included a M72-ChAd vector. The M72-specific CD4+T cell response included predominantly triple (IL2/IFNg/TNFa) and double (IFNg/TNFa) positive cells. In comparison to the benchmark, a reduced level of IL2/TNFa and increased level of IFNg/TNFa secreting cells were observed in the presence of a M72-ChAd vector. The IL-17 secretion was also assessed at 14dPII both in whole blood and in the lungs. However, the detected levels were extremely low across all conditions.
[0524] Similar M72-specific CD8 T cell cytokine profiles were observed across all positive groups in whole blood at 14PI (FIGS. 17 and 18), at 14PII (FIGS. 19 and 20) as well as in the lungs at 14PII (FIGS. 21 and 22). The M72-specific CD8 T cell responses were mostly composed of double (IFNg/TNFa) and single (IFNg only) producing CD8 T cells. Low levels of IL2/INFg/TNFa and very low levels of TNFa producing CD8+T cell were also detected.
[0525] Taken together, the vaccination strategy did not notably impact the cytokine profile of M72 specific CD8 T cell.
Antibody Responses
[0526] A. Anti-M72 Ig Tot Serology
[0527] As shown in FIG. 23, the anti-M72 Ig Tot serology was highly variable across all groups and non-responders were observed with all vaccination strategies except in the combined approach given in co-administration. As for the T cell response, the M72-ChAd3 seems more potent than the M72-ChAd63 at inducing an immune response as the number of non-responding animals is increased when M72-ChAd63 is used.
[0528] Throughout the specification and the claims which follow, unless the context requires otherwise, the word `comprise`, and variations such as `comprises` and `comprising`, will be understood to imply the inclusion of a stated integer, step, group of integers or group of steps but not to the exclusion of any other integer, step, group of integers or group of steps.
[0529] All documents referred to herein, including patents and patent applications, are incorporated by reference in their entirety.
Sequence CWU
1
1
221391PRTMycobacterium tuberculosis 1Met Val Asp Phe Gly Ala Leu Pro Pro
Glu Ile Asn Ser Ala Arg Met1 5 10
15Tyr Ala Gly Pro Gly Ser Ala Ser Leu Val Ala Ala Ala Gln Met
Trp 20 25 30Asp Ser Val Ala
Ser Asp Leu Phe Ser Ala Ala Ser Ala Phe Gln Ser 35
40 45Val Val Trp Gly Leu Thr Val Gly Ser Trp Ile Gly
Ser Ser Ala Gly 50 55 60Leu Met Val
Ala Ala Ala Ser Pro Tyr Val Ala Trp Met Ser Val Thr65 70
75 80Ala Gly Gln Ala Glu Leu Thr Ala
Ala Gln Val Arg Val Ala Ala Ala 85 90
95Ala Tyr Glu Thr Ala Tyr Gly Leu Thr Val Pro Pro Pro Val
Ile Ala 100 105 110Glu Asn Arg
Ala Glu Leu Met Ile Leu Ile Ala Thr Asn Leu Leu Gly 115
120 125Gln Asn Thr Pro Ala Ile Ala Val Asn Glu Ala
Glu Tyr Gly Glu Met 130 135 140Trp Ala
Gln Asp Ala Ala Ala Met Phe Gly Tyr Ala Ala Ala Thr Ala145
150 155 160Thr Ala Thr Ala Thr Leu Leu
Pro Phe Glu Glu Ala Pro Glu Met Thr 165
170 175Ser Ala Gly Gly Leu Leu Glu Gln Ala Ala Ala Val
Glu Glu Ala Ser 180 185 190Asp
Thr Ala Ala Ala Asn Gln Leu Met Asn Asn Val Pro Gln Ala Leu 195
200 205Gln Gln Leu Ala Gln Pro Thr Gln Gly
Thr Thr Pro Ser Ser Lys Leu 210 215
220Gly Gly Leu Trp Lys Thr Val Ser Pro His Arg Ser Pro Ile Ser Asn225
230 235 240Met Val Ser Met
Ala Asn Asn His Met Ser Met Thr Asn Ser Gly Val 245
250 255Ser Met Thr Asn Thr Leu Ser Ser Met Leu
Lys Gly Phe Ala Pro Ala 260 265
270Ala Ala Ala Gln Ala Val Gln Thr Ala Ala Gln Asn Gly Val Arg Ala
275 280 285Met Ser Ser Leu Gly Ser Ser
Leu Gly Ser Ser Gly Leu Gly Gly Gly 290 295
300Val Ala Ala Asn Leu Gly Arg Ala Ala Ser Val Gly Ser Leu Ser
Val305 310 315 320Pro Gln
Ala Trp Ala Ala Ala Asn Gln Ala Val Thr Pro Ala Ala Arg
325 330 335Ala Leu Pro Leu Thr Ser Leu
Thr Ser Ala Ala Glu Arg Gly Pro Gly 340 345
350Gln Met Leu Gly Gly Leu Pro Val Gly Gln Met Gly Ala Arg
Ala Gly 355 360 365Gly Gly Leu Ser
Gly Val Leu Arg Val Pro Pro Arg Pro Tyr Val Met 370
375 380Pro His Ser Pro Ala Ala Gly385
3902391PRTMycobacterium tuberculosis 2Met Val Asp Phe Gly Ala Leu Pro Pro
Glu Ile Asn Ser Ala Arg Met1 5 10
15Tyr Ala Gly Pro Gly Ser Ala Ser Leu Val Ala Ala Ala Lys Met
Trp 20 25 30Asp Ser Val Ala
Ser Asp Leu Phe Ser Ala Ala Ser Ala Phe Gln Ser 35
40 45Val Val Trp Gly Leu Thr Val Gly Ser Trp Ile Gly
Ser Ser Ala Gly 50 55 60Leu Met Val
Ala Ala Ala Ser Pro Tyr Val Ala Trp Met Ser Val Thr65 70
75 80Ala Gly Gln Ala Glu Leu Thr Ala
Ala Gln Val Arg Val Ala Ala Ala 85 90
95Ala Tyr Glu Thr Ala Tyr Gly Leu Thr Val Pro Pro Pro Val
Ile Ala 100 105 110Glu Asn Arg
Ala Glu Leu Met Ile Leu Ile Ala Thr Asn Leu Leu Gly 115
120 125Gln Asn Thr Pro Ala Ile Ala Val Asn Glu Ala
Glu Tyr Gly Glu Met 130 135 140Trp Ala
Gln Asp Ala Ala Ala Met Phe Gly Tyr Ala Ala Ala Thr Ala145
150 155 160Thr Ala Thr Ala Thr Leu Leu
Pro Phe Glu Glu Ala Pro Glu Met Thr 165
170 175Ser Ala Gly Gly Leu Leu Glu Gln Ala Ala Ala Val
Glu Glu Ala Ser 180 185 190Asp
Thr Ala Ala Ala Asn Gln Leu Met Asn Asn Val Pro Gln Ala Leu 195
200 205Gln Gln Leu Ala Gln Pro Thr Gln Gly
Thr Thr Pro Ser Ser Lys Leu 210 215
220Gly Gly Leu Trp Lys Thr Val Ser Pro His Arg Ser Pro Ile Ser Asn225
230 235 240Met Val Ser Met
Ala Asn Asn His Met Ser Met Thr Asn Ser Gly Val 245
250 255Ser Met Thr Asn Thr Leu Ser Ser Met Leu
Lys Gly Phe Ala Pro Ala 260 265
270Ala Ala Ala Gln Ala Val Gln Thr Ala Ala Gln Asn Gly Val Arg Ala
275 280 285Met Ser Ser Leu Gly Ser Ser
Leu Gly Ser Ser Gly Leu Gly Gly Gly 290 295
300Val Ala Ala Asn Leu Gly Arg Ala Ala Ser Val Gly Ser Leu Ser
Val305 310 315 320Pro Gln
Ala Trp Ala Ala Ala Asn Gln Ala Val Thr Pro Ala Ala Arg
325 330 335Ala Leu Pro Leu Thr Ser Leu
Thr Ser Ala Ala Glu Arg Gly Pro Gly 340 345
350Gln Met Leu Gly Gly Leu Pro Val Gly Gln Met Gly Ala Arg
Ala Gly 355 360 365Gly Gly Leu Ser
Gly Val Leu Arg Val Pro Pro Arg Pro Tyr Val Met 370
375 380Pro His Ser Pro Ala Ala Gly385
3903323PRTMycobacterium tuberculosis 3Ala Pro Pro Ala Leu Ser Gln Asp Arg
Phe Ala Asp Phe Pro Ala Leu1 5 10
15Pro Leu Asp Pro Ser Ala Met Val Ala Gln Val Gly Pro Gln Val
Val 20 25 30Asn Ile Asn Thr
Lys Leu Gly Tyr Asn Asn Ala Val Gly Ala Gly Thr 35
40 45Gly Ile Val Ile Asp Pro Asn Gly Val Val Leu Thr
Asn Asn His Val 50 55 60Ile Ala Gly
Ala Thr Asp Ile Asn Ala Phe Ser Val Gly Ser Gly Gln65 70
75 80Thr Tyr Gly Val Asp Val Val Gly
Tyr Asp Arg Thr Gln Asp Val Ala 85 90
95Val Leu Gln Leu Arg Gly Ala Gly Gly Leu Pro Ser Ala Ala
Ile Gly 100 105 110Gly Gly Val
Ala Val Gly Glu Pro Val Val Ala Met Gly Asn Ser Gly 115
120 125Gly Gln Gly Gly Thr Pro Arg Ala Val Pro Gly
Arg Val Val Ala Leu 130 135 140Gly Gln
Thr Val Gln Ala Ser Asp Ser Leu Thr Gly Ala Glu Glu Thr145
150 155 160Leu Asn Gly Leu Ile Gln Phe
Asp Ala Ala Ile Gln Pro Gly Asp Ser 165
170 175Gly Gly Pro Val Val Asn Gly Leu Gly Gln Val Val
Gly Met Asn Thr 180 185 190Ala
Ala Ser Asp Asn Phe Gln Leu Ser Gln Gly Gly Gln Gly Phe Ala 195
200 205Ile Pro Ile Gly Gln Ala Met Ala Ile
Ala Gly Gln Ile Arg Ser Gly 210 215
220Gly Gly Ser Pro Thr Val His Ile Gly Pro Thr Ala Phe Leu Gly Leu225
230 235 240Gly Val Val Asp
Asn Asn Gly Asn Gly Ala Arg Val Gln Arg Val Val 245
250 255Gly Ser Ala Pro Ala Ala Ser Leu Gly Ile
Ser Thr Gly Asp Val Ile 260 265
270Thr Ala Val Asp Gly Ala Pro Ile Asn Ser Ala Thr Ala Met Ala Asp
275 280 285Ala Leu Asn Gly His His Pro
Gly Asp Val Ile Ser Val Thr Trp Gln 290 295
300Thr Lys Ser Gly Gly Thr Arg Thr Gly Asn Val Thr Leu Ala Glu
Gly305 310 315 320Pro Pro
Ala4725PRTArtificial SequenceM72 2-hisMISC_FEATURE(1)..(1)Start
codonMISC_FEATURE(2)..(3)His residues (improved expression in E.
coli)MISC_FEATURE(4)..(135)Rv0125
derivedMISC_FEATURE(136)..(137)LinkersMISC_FEATURE(138)..(528)Rv1196
derivedMISC_FEATURE(529)..(530)LinkersMISC_FEATURE(529)..(530)LinkerMISC_-
FEATURE(531)..(725)Rv0125 derived 4Met His His Thr Ala Ala Ser Asp Asn Phe
Gln Leu Ser Gln Gly Gly1 5 10
15Gln Gly Phe Ala Ile Pro Ile Gly Gln Ala Met Ala Ile Ala Gly Gln
20 25 30Ile Arg Ser Gly Gly Gly
Ser Pro Thr Val His Ile Gly Pro Thr Ala 35 40
45Phe Leu Gly Leu Gly Val Val Asp Asn Asn Gly Asn Gly Ala
Arg Val 50 55 60Gln Arg Val Val Gly
Ser Ala Pro Ala Ala Ser Leu Gly Ile Ser Thr65 70
75 80Gly Asp Val Ile Thr Ala Val Asp Gly Ala
Pro Ile Asn Ser Ala Thr 85 90
95Ala Met Ala Asp Ala Leu Asn Gly His His Pro Gly Asp Val Ile Ser
100 105 110Val Thr Trp Gln Thr
Lys Ser Gly Gly Thr Arg Thr Gly Asn Val Thr 115
120 125Leu Ala Glu Gly Pro Pro Ala Glu Phe Met Val Asp
Phe Gly Ala Leu 130 135 140Pro Pro Glu
Ile Asn Ser Ala Arg Met Tyr Ala Gly Pro Gly Ser Ala145
150 155 160Ser Leu Val Ala Ala Ala Gln
Met Trp Asp Ser Val Ala Ser Asp Leu 165
170 175Phe Ser Ala Ala Ser Ala Phe Gln Ser Val Val Trp
Gly Leu Thr Val 180 185 190Gly
Ser Trp Ile Gly Ser Ser Ala Gly Leu Met Val Ala Ala Ala Ser 195
200 205Pro Tyr Val Ala Trp Met Ser Val Thr
Ala Gly Gln Ala Glu Leu Thr 210 215
220Ala Ala Gln Val Arg Val Ala Ala Ala Ala Tyr Glu Thr Ala Tyr Gly225
230 235 240Leu Thr Val Pro
Pro Pro Val Ile Ala Glu Asn Arg Ala Glu Leu Met 245
250 255Ile Leu Ile Ala Thr Asn Leu Leu Gly Gln
Asn Thr Pro Ala Ile Ala 260 265
270Val Asn Glu Ala Glu Tyr Gly Glu Met Trp Ala Gln Asp Ala Ala Ala
275 280 285Met Phe Gly Tyr Ala Ala Ala
Thr Ala Thr Ala Thr Ala Thr Leu Leu 290 295
300Pro Phe Glu Glu Ala Pro Glu Met Thr Ser Ala Gly Gly Leu Leu
Glu305 310 315 320Gln Ala
Ala Ala Val Glu Glu Ala Ser Asp Thr Ala Ala Ala Asn Gln
325 330 335Leu Met Asn Asn Val Pro Gln
Ala Leu Gln Gln Leu Ala Gln Pro Thr 340 345
350Gln Gly Thr Thr Pro Ser Ser Lys Leu Gly Gly Leu Trp Lys
Thr Val 355 360 365Ser Pro His Arg
Ser Pro Ile Ser Asn Met Val Ser Met Ala Asn Asn 370
375 380His Met Ser Met Thr Asn Ser Gly Val Ser Met Thr
Asn Thr Leu Ser385 390 395
400Ser Met Leu Lys Gly Phe Ala Pro Ala Ala Ala Ala Gln Ala Val Gln
405 410 415Thr Ala Ala Gln Asn
Gly Val Arg Ala Met Ser Ser Leu Gly Ser Ser 420
425 430Leu Gly Ser Ser Gly Leu Gly Gly Gly Val Ala Ala
Asn Leu Gly Arg 435 440 445Ala Ala
Ser Val Gly Ser Leu Ser Val Pro Gln Ala Trp Ala Ala Ala 450
455 460Asn Gln Ala Val Thr Pro Ala Ala Arg Ala Leu
Pro Leu Thr Ser Leu465 470 475
480Thr Ser Ala Ala Glu Arg Gly Pro Gly Gln Met Leu Gly Gly Leu Pro
485 490 495Val Gly Gln Met
Gly Ala Arg Ala Gly Gly Gly Leu Ser Gly Val Leu 500
505 510Arg Val Pro Pro Arg Pro Tyr Val Met Pro His
Ser Pro Ala Ala Gly 515 520 525Asp
Ile Ala Pro Pro Ala Leu Ser Gln Asp Arg Phe Ala Asp Phe Pro 530
535 540Ala Leu Pro Leu Asp Pro Ser Ala Met Val
Ala Gln Val Gly Pro Gln545 550 555
560Val Val Asn Ile Asn Thr Lys Leu Gly Tyr Asn Asn Ala Val Gly
Ala 565 570 575Gly Thr Gly
Ile Val Ile Asp Pro Asn Gly Val Val Leu Thr Asn Asn 580
585 590His Val Ile Ala Gly Ala Thr Asp Ile Asn
Ala Phe Ser Val Gly Ser 595 600
605Gly Gln Thr Tyr Gly Val Asp Val Val Gly Tyr Asp Arg Thr Gln Asp 610
615 620Val Ala Val Leu Gln Leu Arg Gly
Ala Gly Gly Leu Pro Ser Ala Ala625 630
635 640Ile Gly Gly Gly Val Ala Val Gly Glu Pro Val Val
Ala Met Gly Asn 645 650
655Ser Gly Gly Gln Gly Gly Thr Pro Arg Ala Val Pro Gly Arg Val Val
660 665 670Ala Leu Gly Gln Thr Val
Gln Ala Ser Asp Ser Leu Thr Gly Ala Glu 675 680
685Glu Thr Leu Asn Gly Leu Ile Gln Phe Asp Ala Ala Ile Gln
Pro Gly 690 695 700Asp Ala Gly Gly Pro
Val Val Asn Gly Leu Gly Gln Val Val Gly Met705 710
715 720Asn Thr Ala Ala Ser
72552178DNAArtificial SequenceM72 2-his 5atgcatcaca cggccgcgtc cgataacttc
cagctgtccc agggtgggca gggattcgcc 60attccgatcg ggcaggcgat ggcgatcgcg
ggccagatcc gatcgggtgg ggggtcaccc 120accgttcata tcgggcctac cgccttcctc
ggcttgggtg ttgtcgacaa caacggcaac 180ggcgcacgag tccaacgcgt ggtcgggagc
gctccggcgg caagtctcgg catctccacc 240ggcgacgtga tcaccgcggt cgacggcgct
ccgatcaact cggccaccgc gatggcggac 300gcgcttaacg ggcatcatcc cggtgacgtc
atctcggtga cctggcaaac caagtcgggc 360ggcacgcgta cagggaacgt gacattggcc
gagggacccc cggccgaatt catggtggat 420ttcggggcgt taccaccgga gatcaactcc
gcgaggatgt acgccggccc gggttcggcc 480tcgctggtgg ccgcggctca gatgtgggac
agcgtggcga gtgacctgtt ttcggccgcg 540tcggcgtttc agtcggtggt ctggggtctg
acggtggggt cgtggatagg ttcgtcggcg 600ggtctgatgg tggcggcggc ctcgccgtat
gtggcgtgga tgagcgtcac cgcggggcag 660gccgagctga ccgccgccca ggtccgggtt
gctgcggcgg cctacgagac ggcgtatggg 720ctgacggtgc ccccgccggt gatcgccgag
aaccgtgctg aactgatgat tctgatagcg 780accaacctct tggggcaaaa caccccggcg
atcgcggtca acgaggccga atacggcgag 840atgtgggccc aagacgccgc cgcgatgttt
ggctacgccg cggcgacggc gacggcgacg 900gcgacgttgc tgccgttcga ggaggcgccg
gagatgacca gcgcgggtgg gctcctcgag 960caggccgccg cggtcgagga ggcctccgac
accgccgcgg cgaaccagtt gatgaacaat 1020gtgccccagg cgctgcaaca gctggcccag
cccacgcagg gcaccacgcc ttcttccaag 1080ctgggtggcc tgtggaagac ggtctcgccg
catcggtcgc cgatcagcaa catggtgtcg 1140atggccaaca accacatgtc gatgaccaac
tcgggtgtgt cgatgaccaa caccttgagc 1200tcgatgttga agggctttgc tccggcggcg
gccgcccagg ccgtgcaaac cgcggcgcaa 1260aacggggtcc gggcgatgag ctcgctgggc
agctcgctgg gttcttcggg tctgggcggt 1320ggggtggccg ccaacttggg tcgggcggcc
tcggtcggtt cgttgtcggt gccgcaggcc 1380tgggccgcgg ccaaccaggc agtcaccccg
gcggcgcggg cgctgccgct gaccagcctg 1440accagcgccg cggaaagagg gcccgggcag
atgctgggcg ggctgccggt ggggcagatg 1500ggcgccaggg ccggtggtgg gctcagtggt
gtgctgcgtg ttccgccgcg accctatgtg 1560atgccgcatt ctccggcagc cggcgatatc
gccccgccgg ccttgtcgca ggaccggttc 1620gccgacttcc ccgcgctgcc cctcgacccg
tccgcgatgg tcgcccaagt ggggccacag 1680gtggtcaaca tcaacaccaa actgggctac
aacaacgccg tgggcgccgg gaccggcatc 1740gtcatcgatc ccaacggtgt cgtgctgacc
aacaaccacg tgatcgcggg cgccaccgac 1800atcaatgcgt tcagcgtcgg ctccggccaa
acctacggcg tcgatgtggt cgggtatgac 1860cgcacccagg atgtcgcggt gctgcagctg
cgcggtgccg gtggcctgcc gtcggcggcg 1920atcggtggcg gcgtcgcggt tggtgagccc
gtcgtcgcga tgggcaacag cggtgggcag 1980ggcggaacgc cccgtgcggt gcctggcagg
gtggtcgcgc tcggccaaac cgtgcaggcg 2040tcggattcgc tgaccggtgc cgaagagaca
ttgaacgggt tgatccagtt cgatgccgcg 2100atccagcccg gtgatgcggg cgggcccgtc
gtcaacggcc taggacaggt ggtcggtatg 2160aacacggccg cgtcctag
21786723PRTArtificial SequenceM72 no his
6Met Thr Ala Ala Ser Asp Asn Phe Gln Leu Ser Gln Gly Gly Gln Gly1
5 10 15Phe Ala Ile Pro Ile Gly
Gln Ala Met Ala Ile Ala Gly Gln Ile Arg 20 25
30Ser Gly Gly Gly Ser Pro Thr Val His Ile Gly Pro Thr
Ala Phe Leu 35 40 45Gly Leu Gly
Val Val Asp Asn Asn Gly Asn Gly Ala Arg Val Gln Arg 50
55 60Val Val Gly Ser Ala Pro Ala Ala Ser Leu Gly Ile
Ser Thr Gly Asp65 70 75
80Val Ile Thr Ala Val Asp Gly Ala Pro Ile Asn Ser Ala Thr Ala Met
85 90 95Ala Asp Ala Leu Asn Gly
His His Pro Gly Asp Val Ile Ser Val Thr 100
105 110Trp Gln Thr Lys Ser Gly Gly Thr Arg Thr Gly Asn
Val Thr Leu Ala 115 120 125Glu Gly
Pro Pro Ala Glu Phe Met Val Asp Phe Gly Ala Leu Pro Pro 130
135 140Glu Ile Asn Ser Ala Arg Met Tyr Ala Gly Pro
Gly Ser Ala Ser Leu145 150 155
160Val Ala Ala Ala Gln Met Trp Asp Ser Val Ala Ser Asp Leu Phe Ser
165 170 175Ala Ala Ser Ala
Phe Gln Ser Val Val Trp Gly Leu Thr Val Gly Ser 180
185 190Trp Ile Gly Ser Ser Ala Gly Leu Met Val Ala
Ala Ala Ser Pro Tyr 195 200 205Val
Ala Trp Met Ser Val Thr Ala Gly Gln Ala Glu Leu Thr Ala Ala 210
215 220Gln Val Arg Val Ala Ala Ala Ala Tyr Glu
Thr Ala Tyr Gly Leu Thr225 230 235
240Val Pro Pro Pro Val Ile Ala Glu Asn Arg Ala Glu Leu Met Ile
Leu 245 250 255Ile Ala Thr
Asn Leu Leu Gly Gln Asn Thr Pro Ala Ile Ala Val Asn 260
265 270Glu Ala Glu Tyr Gly Glu Met Trp Ala Gln
Asp Ala Ala Ala Met Phe 275 280
285Gly Tyr Ala Ala Ala Thr Ala Thr Ala Thr Ala Thr Leu Leu Pro Phe 290
295 300Glu Glu Ala Pro Glu Met Thr Ser
Ala Gly Gly Leu Leu Glu Gln Ala305 310
315 320Ala Ala Val Glu Glu Ala Ser Asp Thr Ala Ala Ala
Asn Gln Leu Met 325 330
335Asn Asn Val Pro Gln Ala Leu Gln Gln Leu Ala Gln Pro Thr Gln Gly
340 345 350Thr Thr Pro Ser Ser Lys
Leu Gly Gly Leu Trp Lys Thr Val Ser Pro 355 360
365His Arg Ser Pro Ile Ser Asn Met Val Ser Met Ala Asn Asn
His Met 370 375 380Ser Met Thr Asn Ser
Gly Val Ser Met Thr Asn Thr Leu Ser Ser Met385 390
395 400Leu Lys Gly Phe Ala Pro Ala Ala Ala Ala
Gln Ala Val Gln Thr Ala 405 410
415Ala Gln Asn Gly Val Arg Ala Met Ser Ser Leu Gly Ser Ser Leu Gly
420 425 430Ser Ser Gly Leu Gly
Gly Gly Val Ala Ala Asn Leu Gly Arg Ala Ala 435
440 445Ser Val Gly Ser Leu Ser Val Pro Gln Ala Trp Ala
Ala Ala Asn Gln 450 455 460Ala Val Thr
Pro Ala Ala Arg Ala Leu Pro Leu Thr Ser Leu Thr Ser465
470 475 480Ala Ala Glu Arg Gly Pro Gly
Gln Met Leu Gly Gly Leu Pro Val Gly 485
490 495Gln Met Gly Ala Arg Ala Gly Gly Gly Leu Ser Gly
Val Leu Arg Val 500 505 510Pro
Pro Arg Pro Tyr Val Met Pro His Ser Pro Ala Ala Gly Asp Ile 515
520 525Ala Pro Pro Ala Leu Ser Gln Asp Arg
Phe Ala Asp Phe Pro Ala Leu 530 535
540Pro Leu Asp Pro Ser Ala Met Val Ala Gln Val Gly Pro Gln Val Val545
550 555 560Asn Ile Asn Thr
Lys Leu Gly Tyr Asn Asn Ala Val Gly Ala Gly Thr 565
570 575Gly Ile Val Ile Asp Pro Asn Gly Val Val
Leu Thr Asn Asn His Val 580 585
590Ile Ala Gly Ala Thr Asp Ile Asn Ala Phe Ser Val Gly Ser Gly Gln
595 600 605Thr Tyr Gly Val Asp Val Val
Gly Tyr Asp Arg Thr Gln Asp Val Ala 610 615
620Val Leu Gln Leu Arg Gly Ala Gly Gly Leu Pro Ser Ala Ala Ile
Gly625 630 635 640Gly Gly
Val Ala Val Gly Glu Pro Val Val Ala Met Gly Asn Ser Gly
645 650 655Gly Gln Gly Gly Thr Pro Arg
Ala Val Pro Gly Arg Val Val Ala Leu 660 665
670Gly Gln Thr Val Gln Ala Ser Asp Ser Leu Thr Gly Ala Glu
Glu Thr 675 680 685Leu Asn Gly Leu
Ile Gln Phe Asp Ala Ala Ile Gln Pro Gly Asp Ala 690
695 700Gly Gly Pro Val Val Asn Gly Leu Gly Gln Val Val
Gly Met Asn Thr705 710 715
720Ala Ala Ser72172DNAArtificial SequenceM72 no his 7atgacggccg
cgtccgataa cttccagctg tcccagggtg ggcagggatt cgccattccg 60atcgggcagg
cgatggcgat cgcgggccag atccgatcgg gtggggggtc acccaccgtt 120catatcgggc
ctaccgcctt cctcggcttg ggtgttgtcg acaacaacgg caacggcgca 180cgagtccaac
gcgtggtcgg gagcgctccg gcggcaagtc tcggcatctc caccggcgac 240gtgatcaccg
cggtcgacgg cgctccgatc aactcggcca ccgcgatggc ggacgcgctt 300aacgggcatc
atcccggtga cgtcatctcg gtgacctggc aaaccaagtc gggcggcacg 360cgtacaggga
acgtgacatt ggccgaggga cccccggccg aattcatggt ggatttcggg 420gcgttaccac
cggagatcaa ctccgcgagg atgtacgccg gcccgggttc ggcctcgctg 480gtggccgcgg
ctcagatgtg ggacagcgtg gcgagtgacc tgttttcggc cgcgtcggcg 540tttcagtcgg
tggtctgggg tctgacggtg gggtcgtgga taggttcgtc ggcgggtctg 600atggtggcgg
cggcctcgcc gtatgtggcg tggatgagcg tcaccgcggg gcaggccgag 660ctgaccgccg
cccaggtccg ggttgctgcg gcggcctacg agacggcgta tgggctgacg 720gtgcccccgc
cggtgatcgc cgagaaccgt gctgaactga tgattctgat agcgaccaac 780ctcttggggc
aaaacacccc ggcgatcgcg gtcaacgagg ccgaatacgg cgagatgtgg 840gcccaagacg
ccgccgcgat gtttggctac gccgcggcga cggcgacggc gacggcgacg 900ttgctgccgt
tcgaggaggc gccggagatg accagcgcgg gtgggctcct cgagcaggcc 960gccgcggtcg
aggaggcctc cgacaccgcc gcggcgaacc agttgatgaa caatgtgccc 1020caggcgctgc
aacagctggc ccagcccacg cagggcacca cgccttcttc caagctgggt 1080ggcctgtgga
agacggtctc gccgcatcgg tcgccgatca gcaacatggt gtcgatggcc 1140aacaaccaca
tgtcgatgac caactcgggt gtgtcgatga ccaacacctt gagctcgatg 1200ttgaagggct
ttgctccggc ggcggccgcc caggccgtgc aaaccgcggc gcaaaacggg 1260gtccgggcga
tgagctcgct gggcagctcg ctgggttctt cgggtctggg cggtggggtg 1320gccgccaact
tgggtcgggc ggcctcggtc ggttcgttgt cggtgccgca ggcctgggcc 1380gcggccaacc
aggcagtcac cccggcggcg cgggcgctgc cgctgaccag cctgaccagc 1440gccgcggaaa
gagggcccgg gcagatgctg ggcgggctgc cggtggggca gatgggcgcc 1500agggccggtg
gtgggctcag tggtgtgctg cgtgttccgc cgcgacccta tgtgatgccg 1560cattctccgg
cagccggcga tatcgccccg ccggccttgt cgcaggaccg gttcgccgac 1620ttccccgcgc
tgcccctcga cccgtccgcg atggtcgccc aagtggggcc acaggtggtc 1680aacatcaaca
ccaaactggg ctacaacaac gccgtgggcg ccgggaccgg catcgtcatc 1740gatcccaacg
gtgtcgtgct gaccaacaac cacgtgatcg cgggcgccac cgacatcaat 1800gcgttcagcg
tcggctccgg ccaaacctac ggcgtcgatg tggtcgggta tgaccgcacc 1860caggatgtcg
cggtgctgca gctgcgcggt gccggtggcc tgccgtcggc ggcgatcggt 1920ggcggcgtcg
cggttggtga gcccgtcgtc gcgatgggca acagcggtgg gcagggcgga 1980acgccccgtg
cggtgcctgg cagggtggtc gcgctcggcc aaaccgtgca ggcgtcggat 2040tcgctgaccg
gtgccgaaga gacattgaac gggttgatcc agttcgatgc cgcgatccag 2100cccggtgatg
cgggcgggcc cgtcgtcaac ggcctaggac aggtggtcgg tatgaacacg 2160gccgcgtcct
ag
217282172DNAArtificial SequenceM72 no his human optimised 8atgaccgccg
ccagcgacaa cttccagctg tctcagggcg gccagggctt cgccatccct 60atcggccaag
ctatggccat tgctggacag atcagaagcg gcggaggcag ccctaccgtg 120catatcggcc
ctaccgcctt cctgggcctg ggcgtggtgg acaacaacgg caacggcgcc 180agagtgcagc
gggtggtcgg atctgcccct gccgcaagcc tgggcatcag caccggggat 240gtgatcaccg
ccgtggatgg cgcccctatc aacagcgcca cagccatggc cgacgccctg 300aatggacacc
accccggcga cgtgatcagc gtgacctggc agaccaagag cggaggcacc 360agaaccggca
acgtgacact ggccgaggga cctcccgccg agttcatggt ggatttcggc 420gccctgcccc
ccgagatcaa ctccgccagg atgtatgccg gccctggcag cgcctctctg 480gtggccgctg
ctcagatgtg ggacagcgtg gccagcgatc tgttcagcgc cgcctccgcc 540ttccagtccg
tggtctgggg cctgaccgtg ggcagctgga tcggaagcag tgccggcctg 600atggtggctg
ccgcctctcc ctacgtggcc tggatgtcag tcacagccgg ccaggccgaa 660ctgactgccg
ctcaagtgcg agtggctgct gctgcctatg agacagccta cggcctgaca 720gtgcccccac
ccgtgatcgc cgagaaccgg gccgagctga tgatcctgat cgccaccaac 780ctgctgggcc
agaacacccc cgccattgcc gtgaacgagg ccgagtacgg cgagatgtgg 840gcccaggacg
ccgctgccat gtttggctat gccgctgcta cagccaccgc cactgccacc 900ctgctgccct
tcgaagaggc ccccgagatg acctctgccg gcggactgct ggaacaggcc 960gctgccgtgg
aagaggccag cgacacagcc gccgctaacc agctgatgaa caacgtgccc 1020caggccctgc
agcagctggc acagcctaca cagggcacca ccccttctag caagctcggc 1080ggcctgtgga
aaaccgtgtc cccccaccgg tcccccatca gcaacatggt gtccatggcc 1140aacaaccaca
tgagcatgac caacagcggc gtgtccatga ccaataccct gagcagcatg 1200ctgaagggct
ttgccccagc cgctgccgct caggctgtgc agacagctgc tcagaatggc 1260gtgcgggcca
tgagcagcct gggcagttcc ctgggcagct ctggactggg agggggcgtg 1320gccgccaatc
tgggcagagc cgctagcgtg ggcagcctgt ctgtgcctca agcctgggct 1380gctgccaatc
aggccgtgac accagccgct agagccctgc ctctgaccag cctgacctct 1440gctgccgaga
ggggccctgg ccagatgctg ggaggactgc ctgtgggcca gatgggagcc 1500agagccggcg
gaggactgag cggcgtgctg agagtgcccc ccagacccta cgtgatgccc 1560cactctcccg
ccgctggcga tattgcccct cccgccctga gccaggacag attcgccgac 1620ttccctgccc
tgcccctgga tccttctgcc atggtggctc aagtgggacc ccaggtggtg 1680aacatcaaca
ccaagctggg ctacaacaac gccgtgggag ccggcaccgg catcgtgatc 1740gaccccaatg
gcgtggtgct gaccaacaat cacgtgatcg ctggcgccac cgacatcaac 1800gccttcagcg
tgggctccgg ccagacctac ggcgtggacg tggtcggata cgaccggacc 1860caggatgtgg
ccgtgctgca gctgagaggc gctggcggac tgccttctgc cgccattgga 1920ggcggagtgg
ccgtgggaga acctgtggtg gccatgggca atagcggcgg acagggcggc 1980acacctagag
ctgtgcctgg aagagtggtg gccctgggac agaccgtgca ggccagcgat 2040agcctgacag
gcgccgagga aaccctgaac ggcctgatcc agttcgacgc cgccatccag 2100cctggggatg
ctggcggacc tgtggtgaac ggactgggcc aggtggtcgg aatgaatacc 2160gccgcctcct
ga 2172937741DNAPan
troglodytes 9catcatcaat aatatacctt attttggatt gaagccaata tgataatgag
atgggcggcg 60cgaggcgggg cgcggggcgg gaggcgggtt tgggggcggg ccggcgggcg
gggcggtgtg 120gcggaagtgg actttgtaag tgtggcggat gtgacttgct agtgccgggc
gcggtaaaag 180tgacgttttc cgtgcgcgac aacgcccccg ggaagtgaca tttttcccgc
ggtttttacc 240ggatgttgta gtgaatttgg gcgtaaccaa gtaagatttg gccattttcg
cgggaaaact 300gaaacgggga agtgaaatct gattaatttt gcgttagtca taccgcgtaa
tatttgtcta 360gggccgaggg actttggccg attacgtgga ggactcgccc aggtgttttt
tgaggtgaat 420ttccgcgttc cgggtcaaag tctccgtttt attattatag tcagctgacg
cggagtgtat 480ttataccctc tgatctcgtc aagaggccac tcttgagtgc cagcgagtag
agttttctcc 540tctgccgctc tccgctccgc tccgctcggc tctgacaccg gggaaaaaat
gagacatttc 600acctacgatg gcggtgtgct caccggccag ctggctgctg aggtcctgga
caccctgatc 660gaggaggtat tggccgataa ttatcctccc tcgactcctt ttgagccacc
tacacttcac 720gaactatacg atctggatgt ggtggggccc agcgatccga acgagcaggc
ggtttccagt 780ttttttccag agtccatgtt gttggccagc caggaggggg tcgaacttga
gacccctcct 840ccgatcgtgg attcccccga tccgccgcag ctgactaggc agcccgagcg
ctgtgcggga 900cctgagacta tgccccagct gctacctgag gtgatcgatc tcacctgtaa
tgagtctggt 960tttccaccca gcgaggatga ggacgaagag ggtgagcagt ttgtgttaga
ttctgtggaa 1020caacccgggc gaggatgcag gtcttgtcaa tatcaccgga aaaacacagg
agactcccag 1080attatgtgtt ctctgtgtta tatgaagatg acctgtatgt ttatttacag
taagtttatc 1140atcggtgggc aggtgggcta tagtgtgggt ggtggtcttt ggggggtttt
ttaatatatg 1200tcaggggtta tgctgaagac ttttttattg tgatttttaa aggtccagtg
tctgagcccg 1260agcaagaacc tgaaccggag cctgagcctt ctcgccccag gagaaagcct
gtaatcttaa 1320ctagacccag cgcaccggta gcgagaggcc tcagcagcgc ggagaccacc
gactccggtg 1380cttcctcatc acccccggag attcaccccc tggtgcccct atgtcccgtt
aagcccgttg 1440ccgtgagagt cagtgggcgg cggtctgctg tggagtgcat tgaggacttg
ctttttgatt 1500cacaggaacc tttggacttg agcttgaaac gccccaggca ttaaacctgg
tcacctggac 1560tgaatgagtt gacgcctatg tttgcttttg aatgacttaa tgtgtataga
taataaagag 1620tgagataatg ttttaattgc atggtgtgtt taacttgggc ggagtctgct
gggtatataa 1680gcttccctgg gctaaacttg gttacacttg acctcatgga ggcctgggag
tgtttggaga 1740actttgccgg agttcgtgcc ttgctggacg agagctctaa caatacctct
tggtggtgga 1800ggtatttgtg gggctctccc cagggcaagt tagtttgtag aatcaaggag
gattacaagt 1860gggaatttga agagcttttg aaatcctgtg gtgagctatt ggattctttg
aatctaggcc 1920accaggctct cttccaggag aaggtcatca ggactttgga tttttccaca
ccggggcgca 1980ttgcagccgc ggttgctttt ctagcttttt tgaaggatag atggagcgaa
gagacccact 2040tgagttcggg ctacgtcctg gattttctgg ccatgcaact gtggagagca
tggatcagac 2100acaagaacag gctgcaactg ttgtcttccg tccgcccgtt gctgattccg
gcggaggagc 2160aacaggccgg gtcagaggac cgggcccgtc gggatccgga ggagagggca
ccgaggccgg 2220gcgagaggag cgcgctgaac ctgggaaccg ggctgagcgg ccatccacat
cgggagtgaa 2280tgtcgggcag gtggtggatc tttttccaga actgcggcgg attttgacta
ttagggagga 2340tgggcaattt gttaagggtc ttaagaggga gaggggggct tctgagcata
acgaggaggc 2400cagtaattta gcttttagct tgatgaccag acaccgtcca gagtgcatca
cttttcagca 2460gattaaggac aattgtgcca atgagttgga tctgttgggt cagaagtata
gcatagagca 2520gctgaccact tactggctgc agccgggtga tgatctggag gaagctatta
gggtgtatgc 2580taaggtggcc ctgcggcccg attgcaagta caagctcaag gggctggtga
atatcaggaa 2640ttgttgctac atttctggca acggggcgga ggtggagata gagaccgaag
acagggtggc 2700tttcagatgc agcatgatga atatgtggcc gggggtgctg ggcatggacg
gggtggtgat 2760tatgaatgtg aggttcacgg ggcccaactt taacggcacg gtgtttttgg
ggaacaccaa 2820cctggtcctg cacggggtga gcttctatgg gtttaacaac acctgtgtgg
aggcctggac 2880cgatgtgaag gtccgcggtt gcgcctttta tggatgttgg aaggccatag
tgagccgccc 2940taagagcagg agttccatta agaaatgctt gtttgagagg tgcaccttgg
ggatcctggc 3000cgagggcaac tgcagggtgc gccacaatgt ggcctccgag tgcggttgct
tcatgctagt 3060caagagcgtg gcggtaatca agcataatat ggtgtgcggc aacagcgagg
acaaggcctc 3120acagatgctg acctgcacgg atggcaactg ccacttgctg aagaccatcc
atgtaaccag 3180ccacagccgg aaggcctggc ccgtgttcga gcacaacttg ctgacccgct
gctccttgca 3240tctgggcaac aggcgggggg tgttcctgcc ctatcaatgc aactttagtc
acaccaagat 3300cttgctagag cccgagagca tgtccaaggt gaacttgaac ggggtgtttg
acatgaccat 3360gaagatctgg aaggtgctga ggtacgacga gaccaggtcc cggtgcagac
cctgcgagtg 3420cgggggcaag catatgagga accagcccgt gatgctggat gtgaccgagg
agctgaggac 3480agaccacttg gttctggcct gcaccagggc cgagtttggt tctagcgatg
aagacacaga 3540ttgaggtggg tgagtgggcg tggcctgggg tggtcatgaa aatatataag
ttgggggtct 3600tagggtctct ttatttgtgt tgcagagacc gccggagcca tgagcgggag
cagcagcagc 3660agcagtagca gcagcgcctt ggatggcagc atcgtgagcc cttatttgac
gacgcggatg 3720ccccactggg ccggggtgcg tcagaatgtg atgggctcca gcatcgacgg
ccgacccgtc 3780ctgcccgcaa attccgccac gctgacctat gcgaccgtcg cggggacgcc
gttggacgcc 3840accgccgccg ccgccgccac cgcagccgcc tcggccgtgc gcagcctggc
cacggacttt 3900gcattcctgg gaccactggc gacaggggct acttctcggg ccgctgctgc
cgccgttcgc 3960gatgacaagc tgaccgccct gctggcgcag ttggatgcgc ttactcggga
actgggtgac 4020ctttctcagc aggtcatggc cctgcgccag caggtctcct ccctgcaagc
tggcgggaat 4080gcttctccca caaatgccgt ttaagataaa taaaaccaga ctctgtttgg
attaaagaaa 4140agtagcaagt gcattgctct ctttatttca taattttccg cgcgcgatag
gccctagacc 4200agcgttctcg gtcgttgagg gtgcggtgta tcttctccag gacgtggtag
aggtggctct 4260ggacgttgag atacatgggc atgagcccgt cccgggggtg gaggtagcac
cactgcagag 4320cttcatgctc cggggtggtg ttgtagatga tccagtcgta gcaggagcgc
tgggcatggt 4380gcctaaaaat gtccttcagc agcaggccga tggccagggg gaggcccttg
gtgtaagtgt 4440ttacaaaacg gttaagttgg gaagggtgca ttcggggaga gatgatgtgc
atcttggact 4500gtatttttag attggcgatg tttccgccca gatcccttct gggattcatg
ttgtgcagga 4560ccaccagtac agtgtatccg gtgcacttgg ggaatttgtc atgcagctta
gagggaaaag 4620cgtggaagaa cttggagacg cccttgtggc ctcccagatt ttccatgcat
tcgtccatga 4680tgatggcaat gggcccgcgg gaggcagctt gggcaaagat atttctgggg
tcgctgacgt 4740cgtagttgtg ttccagggtg aggtcgtcat aggccatttt tacaaagcgc
gggcggaggg 4800tgcccgactg ggggatgatg gtcccctctg gccctggggc gtagttgccc
tcgcagatct 4860gcatttccca ggccttaatc tcggaggggg gaatcatatc cacctgcggg
gcgatgaaga 4920aaacggtttc cggagccggg gagattaact gggatgagag caggtttcta
agcagctgtg 4980attttccaca accggtgggc ccataaataa cacctataac cggttgcagc
tggtagttta 5040gagagctgca gctgccgtcg tcccggagga ggggggccac ctcgttgagc
atgtccctga 5100cgcgcatgtt ctccccgacc agatccgcca gaaggcgctc gccgcccagg
gacagcagct 5160cttgcaagga agcaaagttt ttcagcggct tgaggccgtc cgccgtgggc
atgtttttca 5220gggtctggct cagcagctcc aggcggtccc agagctcggt gacgtgctct
acggcatctc 5280tatccagcat atctcctcgt ttcgcgggtt ggggcgactt tcgctgtagg
gcaccaagcg 5340gtggtcgtcc agcggggcca aagtcatgtc cttccatggg cgcagggtcc
tcgtcagggt 5400ggtctgggtc acggtgaagg ggtgcgctcc gggctgagcg cttgccaagg
tgcgcttgag 5460gctggttctg ctggtgctga agcgctgccg gtcttcgccc tgcgcgtcgg
ccaggtagca 5520tttgaccatg gtgtcatagt ccagcccctc cgcggcgtgt cccttggcgc
gcagcttgcc 5580cttggaggtg gcgccgcacg aggggcagag caggctcttg agcgcgtaga
gcttgggggc 5640gaggaagacc gattcggggg agtaggcgtc cgcgccgcag accccgcaca
cggtctcgca 5700ctccaccagc caggtgagct cggggcgcgc cgggtcaaaa accaggtttc
ccccatgctt 5760tttgatgcgt ttcttacctc gggtctccat gaggtggtgt ccccgctcgg
tgacgaagag 5820gctgtccgtg tctccgtaga ccgacttgag gggtcttttc tccagggggg
tccctcggtc 5880ttcctcgtag aggaactcgg accactctga gacgaaggcc cgcgtccagg
ccaggacgaa 5940ggaggctatg tgggaggggt agcggtcgtt gtccactagg gggtccacct
tctccaaggt 6000gtgaagacac atgtcgcctt cctcggcgtc caggaaggtg attggcttgt
aggtgtaggc 6060cacgtgaccg ggggttcctg acgggggggt ataaaagggg gtgggggcgc
gctcgtcgtc 6120actctcttcc gcatcgctgt ctgcgagggc cagctgctgg ggtgagtatt
ccctctcgaa 6180ggcgggcatg acctccgcgc tgaggttgtc agtttccaaa aacgaggagg
atttgatgtt 6240cacctgtccc gaggtgatac ctttgagggt acccgcgtcc atctggtcag
aaaacacgat 6300ctttttattg tccagcttgg tggcgaacga cccgtagagg gcgttggaga
gcagcttggc 6360gatggagcgc agggtctggt tcttgtccct gtcggcgcgc tccttggccg
cgatgttgag 6420ctgcacgtac tcgcgcgcga cgcagcgcca ctcggggaag acggtggtgc
gctcgtcggg 6480caccaggcgc acgcgccagc cgcggttgtg cagggtgacc aggtccacgc
tggtggcgac 6540ctcgccgcgc aggcgctcgt tggtccagca gagacggccg cccttgcgcg
agcagaaggg 6600gggcaggggg tcgagctggg tctcgtccgg ggggtccgcg tccacggtga
aaaccccggg 6660gcgcaggcgc gcgtcgaagt agtctatctt gcaaccttgc atgtccagcg
cctgctgcca 6720gtcgcgggcg gcgagcgcgc gctcgtaggg gttgagcggc gggccccagg
gcatggggtg 6780ggtgagtgcg gaggcgtaca tgccgcagat gtcatagacg tagaggggct
cccgcaggac 6840cccgatgtag gtggggtagc agcggccgcc gcggatgctg gcgcgcacgt
agtcatacag 6900ctcgtgcgag ggggcgagga ggtcggggcc caggttggtg cgggcggggc
gctccgcgcg 6960gaagacgatc tgcctgaaga tggcatgcga gttggaagag atggtggggc
gctggaagac 7020gttgaagctg gcgtcctgca ggccgacggc gtcgcgcacg aaggaggcgt
aggagtcgcg 7080cagcttgtgt accagctcgg cggtgacctg cacgtcgagc gcgcagtagt
cgagggtctc 7140gcggatgatg tcatatttag cctgcccctt ctttttccac agctcgcggt
tgaggacaaa 7200ctcttcgcgg tctttccagt actcttggat cgggaaaccg tccggttccg
aacggtaaga 7260gcctagcatg tagaactggt tgacggcctg gtaggcgcag cagcccttct
ccacggggag 7320ggcgtaggcc tgcgcggcct tgcggagcga ggtgtgggtc agggcgaagg
tgtccctgac 7380catgactttg aggtactggt gcttgaagtc ggagtcgtcg cagccgcccc
gctcccagag 7440cgagaagtcg gtgcgcttct tggagcgggg gttgggcaga gcgaaggtga
catcgttgaa 7500gaggattttg cccgcgcggg gcatgaagtt gcgggtgatg cggaagggcc
ccggcacttc 7560agagcggttg ttgatgacct gggcggcgag cacgatctcg tcgaagccgt
tgatgttgtg 7620gcccacgatg tagagttcca ggaagcgggg ccggcccttt acggtgggca
gcttctttag 7680ctcttcgtag gtgagctcct cgggcgaggc gaggccgtgc tcggccaggg
cccagtccgc 7740gaggtgcggg ttgtctctga ggaaggactc ccagaggtcg cgggccagga
gggtctgcag 7800gcggtccctg aaggtcctga actggcggcc cacggccatt ttttcggggg
tgatgcagta 7860gaaggtgagg gggtcttgct gccagcggtc ccagtcgagc tgcagggcga
ggtcgcgcgc 7920ggcggtgacc aggcgctcgt cgcccccgaa tttcatgacc agcatgaagg
gcacgagctg 7980ctttccgaag gcccccatcc aagtgtaggt ctctacatcg taggtgacaa
agaggcgctc 8040cgtgcgagga tgcgagccga tcgggaagaa ctggatctcc cgccaccagt
tggaggagtg 8100gctgttgatg tggtggaagt agaagtcccg tcgccgggcc gaacactcgt
gctggctttt 8160gtaaaagcga gcgcagtact ggcagcgctg cacgggctgt acctcctgca
cgagatgcac 8220ctttcgcccg cgcacgagga agccgagggg aaatctgagc cccccgcctg
gctcgcggca 8280tggctggtgc tcttctactt tggatgcgtg tccgtctccg tctggctcct
cgaggggtgt 8340tacggtggag cggaccacca cgccgcgcga gccgcaggtc cagatatcgg
cgcgcggcgg 8400tcggagtttg atgacgacat cgcgcagctg ggagctgtcc atggtctgga
gctcccgcgg 8460cggcggcagg tcagccggga gttcttgcag gttcacctcg cagagtcggg
ccagggcgcg 8520gggcaggtct aggtggtacc tgatctctag gggcgtgttg gtggcggcgt
cgatggcttg 8580caggagcccg catccccggg gggcgacgac ggtgccccgc ggggtggtgg
tggtggtggt 8640ggtggtggtg gtggcggtgc agctcagaag cggtgccgcg ggcgggcccc
cggaggtagg 8700gggggctccg gtcccgccgg caggggcggc agcggcacgt cggcgtggag
cgcgggcagg 8760agttggtgct gtgcccggag gttgctggcg aaggcgacga cgcggcggtt
gatctcctgg 8820atctggcgcc tctgcgtgaa gacgacgggc ccggtgagct tgaacctgaa
agagagttcg 8880acagaatcaa tctcggtgtc attgaccgcg gcctggcgca ggatctcctg
cacgtctccc 8940gagttgtctt ggtaggcgat ctcggccatg aactgctcga tctcttcctc
ctggaggtct 9000ccgcgtccgg cgcgttccac ggtggccgcc aggtcgttgg agatgcgccc
catgagctgc 9060gagaaggcgt tgagtccgcc ctcgttccag actcggctgt agaccacgcc
cccctggtca 9120tcgcgggcgc gcatgaccac ctgcgcgagg ttgagctcca cgtgccgcgc
gaagacggcg 9180tagttgcgca gacgctggaa gaggtagttg agggtggtgg cggtgtgctc
ggccacgaag 9240aagttcatga cccagcggcg caacgtggat tcgttgatgt cccccaaggc
ctccagccgt 9300tccatggcct cgtagaagtc cacggcgaag ttgaaaaact gggagttgcg
cgccgacacg 9360gtcaactcct cctccagaag acggatgagc tcggcgacgg tgtcgcgcac
ctcgcgctcg 9420aaggctatgg ggatctcttc ctccgctagc atcaccacct cctcctcttc
ctcctcttct 9480ggcacttcca tgatggcttc ctcctcttcg gggggcggcg gcggcggcgg
tgggggaggg 9540ggcgctctgc gccggcggcg gcgcaccggg aggcggtcca cgaagcgcgc
gatcatctcc 9600ccgcggcggc ggcgcatggt ctcggtgacg gcgcggccgt tctcccgggg
gcgcagttgg 9660aagacgccgc cggacatctg gtgctggggc gggtggccgt gaggcagcga
aacggcgctg 9720acgatgcatc tcaacaattg ctgcgtaggt acgccgccga gggacctgag
ggagtccata 9780tccaccggat ccgaaaacct ttcgaggaag gcgtctaacc agtcgcagtc
gcaaggtagg 9840ctgagcaccg tggcgggcgg cggggggtgg ggggagtgtc tggcggaggt
gctgctgatg 9900atgtaattga agtaggcgga cttgacacgg cggatggtcg acaggagcac
catgtccttg 9960ggtccggcct gctggatgcg gaggcggtcg gctatgcccc aggcttcgtt
ctggcatcgg 10020cgcaggtcct tgtagtagtc ttgcatgagc ctttccaccg gcacctcttc
tccttcctct 10080tctgcttctt ccatgtctgc ttcggccctg gggcggcgcc gcgcccccct
gccccccatg 10140cgcgtgaccc cgaaccccct gagcggttgg agcagggcca ggtcggcgac
gacgcgctcg 10200gccaggatgg cctgctgcac ctgcgtgagg gtggtttgga agtcatccaa
gtccacgaag 10260cggtggtagg cgcccgtgtt gatggtgtag gtgcagttgg ccatgacgga
ccagttgacg 10320gtctggtggc ccggttgcga catctcggtg tacctgagtc gcgagtaggc
gcgggagtcg 10380aagacgtagt cgttgcaagt ccgcaccagg tactggtagc ccaccaggaa
gtgcggcggc 10440ggctggcggt agaggggcca gcgcagggtg gcgggggctc cgggggccag
gtcttccagc 10500atgaggcggt ggtaggcgta gatgtacctg gacatccagg tgatacccgc
ggcggtggtg 10560gaggcgcgcg ggaagtcgcg cacccggttc cagatgttgc gcaggggcag
aaagtgctcc 10620atggtaggcg tgctctgtcc agtcagacgc gcgcagtcgt tgatactcta
gaccagggaa 10680aacgaaagcc ggtcagcggg cactcttccg tggtctggtg aatagatcgc
aagggtatca 10740tggcggaggg cctcggttcg agccccgggt ccgggccgga cggtccgcca
tgatccacgc 10800ggttaccgcc cgcgtgtcga acccaggtgt gcgacgtcag acaacggtgg
agtgttcctt 10860ttggcgtttt tctggccggg cgccggcgtc gcgtaagaga ctaagccgcg
aaagcgaaag 10920cagtaagtgg ctcgctcccc gtagccggag ggatccttgc taagggttgc
gttgcggcga 10980accccggttc gaatcccgta ctcgggccgg ccggacccgc ggctaaggtg
ttggattggc 11040ctccccctcg tataaagacc ccgcttgcgg attgactccg gacacgggga
cgagcccctt 11100ttatttttgc tttccccaga tgcatccggt gctgcggcag atgcgccccc
cgccccagca 11160gcagcaacaa caccagcaag agcggcagca acagcagcgg gagtcatgca
gggccccctc 11220acccaccctc ggcgggccgg ccacctcggc gtccgcggcc gtgtctggcg
cctgcggcgg 11280cggcgggggg ccggctgacg accccgagga gcccccgcgg cgcagggcca
gacactacct 11340ggacctggag gagggcgagg gcctggcgcg gctgggggcg ccgtctcccg
agcgccaccc 11400gcgggtgcag ctgaagcgcg actcgcgcga ggcgtacgtg cctcggcaga
acctgttcag 11460ggaccgcgcg ggcgaggagc ccgaggagat gcgggacagg aggttcagcg
cagggcggga 11520gctgcggcag gggctgaacc gcgagcggct gctgcgcgag gaggactttg
agcccgacgc 11580gcggacgggg atcagccccg cgcgcgcgca cgtggcggcc gccgacctgg
tgacggcgta 11640cgagcagacg gtgaaccagg agatcaactt ccaaaagagt ttcaacaacc
acgtgcgcac 11700gctggtggcg cgcgaggagg tgaccatcgg gctgatgcac ctgtgggact
ttgtaagcgc 11760gctggtgcag aaccccaaca gcaagcctct gacggcgcag ctgttcctga
tagtgcagca 11820cagcagggac aacgaggcgt ttagggacgc gctgctgaac atcaccgagc
ccgagggtcg 11880gtggctgctg gacctgatta acatcctgca gagcatagtg gtgcaggagc
gcagcctgag 11940cctggccgac aaggtggcgg ccatcaacta ctcgatgctg agcctgggca
agttttacgc 12000gcgcaagatc taccagacgc cgtacgtgcc catagacaag gaggtgaaga
tcgacggttt 12060ttacatgcgc atggcgctga aggtgctcac cctgagcgac gacctgggcg
tgtaccgcaa 12120cgagcgcatc cacaaggccg tgagcgtgag ccggcggcgc gagctgagcg
accgcgagct 12180gatgcacagc ctgcagcggg cgctggcggg cgccggcagc ggcgacaggg
aggcggagtc 12240ctacttcgat gcgggggcgg acctgcgctg ggcgcccagc cggcgggccc
tggaggccgc 12300gggggtccgc gaggactatg acgaggacgg cgaggaggat gaggagtacg
agctagagga 12360gggcgagtac ctggactaaa ccgcgggtgg tgtttccggt agatgcaaga
cccgaacgtg 12420gtggacccgg cgctgcgggc ggctctgcag agccagccgt ccggccttaa
ctcctcagac 12480gactggcgac aggtcatgga ccgcatcatg tcgctgacgg cgcgtaaccc
ggacgcgttc 12540cggcagcagc cgcaggccaa caggctctcc gccatcctgg aggcggtggt
gcctgcgcgc 12600tcgaacccca cgcacgagaa ggtgctggcc atagtgaacg cgctggccga
gaacagggcc 12660atccgcccgg acgaggccgg gctggtgtac gacgcgctgc tgcagcgcgt
ggcccgctac 12720aacagcggca acgtgcagac caacctggac cggctggtgg gggacgtgcg
cgaggcggtg 12780gcgcagcgcg agcgcgcgga tcggcagggc aacctgggct ccatggtggc
gctgaatgcc 12840ttcctgagca cgcagccggc caacgtgccg cgggggcagg aagactacac
caactttgtg 12900agcgcgctgc ggctgatggt gaccgagacc ccccagagcg aggtgtacca
gtcgggcccg 12960gactacttct tccagaccag cagacagggc ctgcagacgg tgaacctgag
ccaggctttc 13020aagaacctgc gggggctgtg gggcgtgaag gcgcccaccg gcgaccgggc
gacggtgtcc 13080agcctgctga cgcccaactc gcgcctgctg ctgctgctga tcgcgccgtt
cacggacagc 13140ggcagcgtgt cccgggacac ctacctgggg cacctgctga ccctgtaccg
cgaggccatc 13200gggcaggcgc aggtggacga gcacaccttc caggagatca ccagcgtgag
ccgcgcgctg 13260gggcaggagg acacgagcag cctggaggcg actctgaact acctgctgac
caaccggcgg 13320cagaagattc cctcgctgca cagcctgacc tccgaggagg agcgcatctt
gcgctacgtg 13380cagcagagcg tgagcctgaa cctgatgcgc gacggggtga cgcccagcgt
ggcgctggac 13440atgaccgcgc gcaacatgga accgggcatg tacgccgcgc accggcctta
catcaaccgc 13500ctgatggact acctgcatcg cgcggcggcc gtgaaccccg agtactttac
caacgccatc 13560ctgaacccgc actggctccc gccgcccggg ttctacagcg ggggcttcga
ggtcccggag 13620gccaacgatg gcttcctgtg ggacgacatg gacgacagcg tgttctcccc
gcggccgcag 13680gcgctggcgg aagcgtccct gctgcgtccc aagaaggagg aggaggaggc
gagtcgccgc 13740cgcggcagca gcggcgtggc ttctctgtcc gagctggggg cggcagccgc
cgcgcgcccc 13800gggtccctgg gcggcagccc ctttccgagc ctggtggggt ctctgcacag
cgagcgcacc 13860acccgccctc ggctgctggg cgaggacgag tacctgaata actccctgct
gcagccggtg 13920cgggagaaaa acctgccccc cgccttcccc aacaacggga tagagagcct
ggtggacaag 13980atgagcagat ggaagaccta tgcgcaggag cacagggacg cgcccgcgct
ccggccgccc 14040acgcggcgcc agcgccacga ccggcagcgg gggctggtgt gggatgacga
ggactccgcg 14100gacgatagca gcgtgctgga cctgggaggg agcggcaacc cgttcgcgca
cctgcgcccc 14160cgcctgggga ggatgtttta aaaaaaaaaa aagcaagaag catgatgcaa
aattaaataa 14220aactcaccaa ggccatggcg accgagcgtt ggtttcttgt gttcccttca
gtatgcggcg 14280cgcggcgatg taccaggagg gacctcctcc ctcttacgag agcgtggtgg
gcgcggcggc 14340ggcggcgccc tcttctccct ttgcgtcgca gctgctggag ccgccgtacg
tgcctccgcg 14400ctacctgcgg cctacggggg ggagaaacag catccgttac tcggagctgg
cgcccctgtt 14460cgacaccacc cgggtgtacc tggtggacaa caagtcggcg gacgtggcct
ccctgaacta 14520ccagaacgac cacagcaatt ttttgaccac ggtcatccag aacaatgact
acagcccgag 14580cgaggccagc acccagacca tcaatctgga tgaccggtcg cactggggcg
gcgacctgaa 14640aaccatcctg cacaccaaca tgcccaacgt gaacgagttc atgttcacca
ataagttcaa 14700ggcgcgggtg atggtgtcgc gctcgcacac caaggaagac cgggtggagc
tgaagtacga 14760gtgggtggag ttcgagctgc cagagggcaa ctactccgag accatgacca
ttgacctgat 14820gaacaacgcg atcgtggagc actatctgaa agtgggcagg caaaacgggg
tcctggagag 14880cgacatcggg gtcaagttcg acaccaggaa cttccgcctg gggctggacc
ccgtgaccgg 14940gctggttatg cccggggtgt acaccaacga ggccttccat cccgacatca
tcctgctgcc 15000cggctgcggg gtggacttca cttacagccg cctgagcaac ctcctgggca
tccgcaagcg 15060gcagcccttc caggagggct tcaggatcac ctacgaggac ctggaggggg
gcaacatccc 15120cgcgctcctc gatgtggagg cctaccagga tagcttgaag gaaaatgagg
cgggacagga 15180ggataccacc cccgccgcct ccgccgccgc cgagcagggc gaggatgctg
ctgacaccgc 15240ggccgcggac ggggcagagg ccgaccccgc tatggtggtg gaggctcccg
agcaggagga 15300ggatatgaat gacagtgcgg tgcgcggaga caccttcgtc acccgggggg
aggaaaagca 15360agcggaggcc gaggccgcgg ccgaggaaaa gcaactggcg gcagcagcgg
cggcggcggc 15420gttggccgcg gcggaggctg agtctgaggg gaccaagccc gccaaggagc
ccgtgattaa 15480gcccctgacc gaagatagca agaagcgcag ttacaacctg ctcaaggaca
gcaccaacac 15540cgcgtaccgc agctggtacc tggcctacaa ctacggcgac ccgtcgacgg
gggtgcgctc 15600ctggaccctg ctgtgcacgc cggacgtgac ctgcggctcg gagcaggtgt
actggtcgct 15660gcccgacatg atgcaagacc ccgtgacctt ccgctccacg cggcaggtca
gcaacttccc 15720ggtggtgggc gccgagctgc tgcccgtgca ctccaagagc ttctacaacg
accaggccgt 15780ctactcccag ctcatccgcc agttcacctc tctgacccac gtgttcaatc
gctttcctga 15840gaaccagatt ctggcgcgcc cgcccgcccc caccatcacc accgtcagtg
aaaacgttcc 15900tgctctcaca gatcacggga cgctaccgct gcgcaacagc atcggaggag
tccagcgagt 15960gaccgttact gacgccagac gccgcacctg cccctacgtt tacaaggcct
tgggcatagt 16020ctcgccgcgc gtcctttcca gccgcacttt ttgagcaaca ccaccatcat
gtccatcctg 16080atctcaccca gcaataactc cggctgggga ctgctgcgcg cgcccagcaa
gatgttcgga 16140ggggcgagga agcgttccga gcagcacccc gtgcgcgtgc gcgggcactt
ccgcgccccc 16200tggggagcgc acaaacgcgg ccgcgcgggg cgcaccaccg tggacgacgc
catcgactcg 16260gtggtggagc aggcgcgcaa ctacaggccc gcggtctcta ccgtggacgc
ggccatccag 16320accgtggtgc ggggcgcgcg gcggtacgcc aagctgaaga gccgccggaa
gcgcgtggcc 16380cgccgccacc gccgccgacc cggggccgcc gccaaacgcg ccgccgcggc
cctgcttcgc 16440cgggccaagc gcacgggccg ccgcgccgcc atgagggccg cgcgccgctt
ggccgccggc 16500atcaccgccg ccaccatggc cccccgtacc cgaagacgcg cggccgccgc
cgccgccgcc 16560gccatcagtg acatggccag caggcgccgg ggcaacgtgt actgggtgcg
cgactcggtg 16620accggcacgc gcgtgcccgt gcgcttccgc cccccgcgga cttgagatga
tgtgaaaaaa 16680caacactgag tctcctgctg ttgtgtgtat cccagcggcg gcggcgcgcg
cagcgtcatg 16740tccaagcgca aaatcaaaga agagatgctc caggtcgtcg cgccggagat
ctatgggccc 16800ccgaagaagg aagagcagga ttcgaagccc cgcaagataa agcgggtcaa
aaagaaaaag 16860aaagatgatg acgatgccga tggggaggtg gagttcctgc gcgccacggc
gcccaggcgc 16920ccggtgcagt ggaagggccg gcgcgtaaag cgcgtcctgc gccccggcac
cgcggtggtc 16980ttcacgcccg gcgagcgctc cacccggact ttcaagcgcg tctatgacga
ggtgtacggc 17040gacgaagacc tgctggagca ggccaacgag cgcttcggag agtttgctta
cgggaagcgt 17100cagcgggcgc tggggaagga ggacctgctg gcgctgccgc tggaccaggg
caaccccacc 17160cccagtctga agcccgtgac cctgcagcag gtgctgccga gcagcgcacc
ctccgaggcg 17220aagcggggtc tgaagcgcga gggcggcgac ctggcgccca ccgtgcagct
catggtgccc 17280aagcggcaga ggctggagga tgtgctggag aaaatgaaag tagaccccgg
tctgcagccg 17340gacatcaggg tccgtcccat caagcaggtg gcgccgggcc tcggcgtgca
gaccgtggac 17400gtggtcatcc ccaccggcaa ctcccccgcc gccaccacca ctaccgctgc
ctccacggac 17460atggagacac agaccgatcc cgccgcagcc gcagccgccg ccgcagccgc
gacctcctcg 17520gcggaggtgc agacggaccc ctggctgccg ccggcgatgt cagctccccg
cgcgcgccgc 17580ggacgcagaa agtacggcgc cgccaacgcg ctcctgcccg agtacgcctt
gcatccttcc 17640atcgcgccca cccccggcta ccgaggctat acctaccgcc cgcgaagagc
caagggttcc 17700acccgccgtc cccgccgacg cgccgccgcc accacccgcc gccgccgccg
cagacgccag 17760cccgcactgg ctccagtctc cgtgaggaga gtggcgcgcg acggacacac
cctggtgctg 17820cccagggcgc gctaccaccc cagcatcgtt taaaagcctg ttgtggttct
tgcagatatg 17880gccctcactt gccgcctccg tttcccggtg ccgggatacc gaggaggaag
atcgcgccgc 17940aggaggggtc tggccggccg cggcctgagc ggaggcagcc gccgcgcgca
ccggcggcga 18000cgcgccacca gccgacgcat gcgcggcggg gtgctgcccc tgttaatccc
cctgatcgcc 18060gcggcgatcg gcgccgtgcc cgggatcgcc tccgtggcct tgcaagcgtc
ccagaggcat 18120tgacagactt gcaaacttgc aaatatggaa aaaaaaaaaa aaccccaata
aaaagtctag 18180actctcacgc tcgcttggtc ctgtgactat tttgtagaat ggaagacatc
aactttgcgt 18240cgctggcccc gcgtcacggc tcgcgcccgt tcctgggaca ctggaacgat
atcggcacca 18300gcaacatgag cggtggcgcc ttcagttggg gctctctgtg gagcggcatt
aaaagtatcg 18360ggtctgccgt taaaaattac ggctcccggg cctggaacag cagcacgggc
cagatgttga 18420gagacaagtt gaaagagcag aacttccagc agaaggtggt ggagggcctg
gcctccggca 18480tcaacggggt ggtggacctg gccaaccagg ccgtgcagaa taaaatcaac
agcagactgg 18540acccccggcc gccggtggag gaggtgccgc cggcgctgga gacggtgtcc
cccgatgggc 18600gtggcgagaa gcgcccgcgg cccgataggg aagagaccac tctggtcacg
cagaccgatg 18660agccgccccc gtatgaggag gccctaaagc aaggtctgcc caccacgcgg
cccatcgcgc 18720ccatggccac cggggtggtg ggccgccaca cccccgccac gctggacttg
cctccgcccg 18780ccgatgtgcc gcagcagcag aaggcggcac agccgggccc gcccgcgacc
gcctcccgtt 18840cctccgccgg tcctctgcgc cgcgcggcca gcggcccccg cgggggggtc
gcgaggcacg 18900gcaactggca gagcacgctg aacagcatcg tgggtctggg ggtgcggtcc
gtgaagcgcc 18960gccgatgcta ctgaatagct tagctaacgt gttgtatgtg tgtatgcgcc
ctatgtcgcc 19020gccagaggag ctgctgagtc gccgccgttc gcgcgcccac caccaccgcc
actccgcccc 19080tcaagatggc gaccccatcg atgatgccgc agtggtcgta catgcacatc
tcgggccagg 19140acgcctcgga gtacctgagc cccgggctgg tgcagttcgc ccgcgccacc
gagagctact 19200tcagcctgag taacaagttt aggaacccca cggtggcgcc cacgcacgat
gtgaccaccg 19260accggtctca gcgcctgacg ctgcggttca ttcccgtgga ccgcgaggac
accgcgtact 19320cgtacaaggc gcggttcacc ctggccgtgg gcgacaaccg cgtgctggac
atggcctcca 19380cctactttga catccgcggg gtgctggacc ggggtcccac tttcaagccc
tactctggca 19440ccgcctacaa ctccctggcc cccaagggcg ctcccaactc ctgcgagtgg
gagcaagagg 19500aaactcaggc agttgaagaa gcagcagaag aggaagaaga agatgctgac
ggtcaagctg 19560aggaagagca agcagctacc aaaaagactc atgtatatgc tcaggctccc
ctttctggcg 19620aaaaaattag taaagatggt ctgcaaatag gaacggacgc tacagctaca
gaacaaaaac 19680ctatttatgc agaccctaca ttccagcccg aaccccaaat cggggagtcc
cagtggaatg 19740aggcagatgc tacagtcgcc ggcggtagag tgctaaagaa atctactccc
atgaaaccat 19800gctatggttc ctatgcaaga cccacaaatg ctaatggagg tcagggtgta
ctaacggcaa 19860atgcccaggg acagctagaa tctcaggttg aaatgcaatt cttttcaact
tctgaaaacg 19920cccgtaacga ggctaacaac attcagccca aattggtgct gtatagtgag
gatgtgcaca 19980tggagacccc ggatacgcac ctttcttaca agcccgcaaa aagcgatgac
aattcaaaaa 20040tcatgctggg tcagcagtcc atgcccaaca gacctaatta catcggcttc
agagacaact 20100ttatcggcct catgtattac aatagcactg gcaacatggg agtgcttgca
ggtcaggcct 20160ctcagttgaa tgcagtggtg gacttgcaag acagaaacac agaactgtcc
taccagctct 20220tgcttgattc catgggtgac agaaccagat acttttccat gtggaatcag
gcagtggaca 20280gttatgaccc agatgttaga attattgaaa atcatggaac tgaagacgag
ctccccaact 20340attgtttccc tctgggtggc ataggggtaa ctgacactta ccaggctgtt
aaaaccaaca 20400atggcaataa cgggggccag gtgacttgga caaaagatga aacttttgca
gatcgcaatg 20460aaataggggt gggaaacaat ttcgctatgg agatcaacct cagtgccaac
ctgtggagaa 20520acttcctgta ctccaacgtg gcgctgtacc taccagacaa gcttaagtac
aacccctcca 20580atgtggacat ctctgacaac cccaacacct acgattacat gaacaagcga
gtggtggccc 20640cggggctggt ggactgctac atcaacctgg gcgcgcgctg gtcgctggac
tacatggaca 20700acgtcaaccc cttcaaccac caccgcaatg cgggcctgcg ctaccgctcc
atgctcctgg 20760gcaacgggcg ctacgtgccc ttccacatcc aggtgcccca gaagttcttt
gccatcaaga 20820acctcctcct cctgccgggc tcctacacct acgagtggaa cttcaggaag
gatgtcaaca 20880tggtcctcca gagctctctg ggtaacgatc tcagggtgga cggggccagc
atcaagttcg 20940agagcatctg cctctacgcc accttcttcc ccatggccca caacacggcc
tccacgctcg 21000aggccatgct caggaacgac accaacgacc agtccttcaa tgactacctt
tccgccgcca 21060acatgctcta ccccataccc gccaacgcca ccaacgtccc catctccatc
ccctcgcgca 21120actgggcggc cttccgcggc tgggccttca cccgcctcaa gaccaaggag
accccctccc 21180tgggctcggg attcgacccc tactacacct actcgggctc tattccctac
ctggacggca 21240ccttctacct caaccacact ttcaagaagg tctcggtcac cttcgactcc
tcggtcagct 21300ggccgggcaa cgaccgtctg ctcaccccca acgagttcga gatcaagcgc
tcggtcgacg 21360gggaaggcta caacgtggcc cagtgcaaca tgaccaagga ctggttcctg
gtccagatgc 21420tggccaacta caacatcggc taccagggct tctacatccc agagagctac
aaggacagga 21480tgtactcctt cttcaggaac ttccagccca tgagccggca ggtggtggac
cagaccaagt 21540acaaggacta ccaggaggtg ggcatcatcc accagcacaa caactcgggc
ttcgtgggct 21600acctcgcccc caccatgcgc gagggacagg cctaccccgc caacttcccc
tacccgctca 21660taggcaagac cgcggtcgac agcatcaccc agaaaaagtt cctctgcgac
cgcaccctct 21720ggcgcatccc cttctccagc aacttcatgt ccatgggtgc gctctcggac
ctgggccaga 21780acttgctcta cgccaactcc gcccacgccc tcgacatgac cttcgaggtc
gaccccatgg 21840acgagcccac ccttctctat gttctgttcg aagtctttga cgtggtccgg
gtccaccagc 21900cgcaccgcgg cgtcatcgag accgtgtacc tgcgtacgcc cttctcggcc
ggcaacgcca 21960ccacctaaag aagcaagccg cagtcatcgc cgcctgcatg ccgtcgggtt
ccaccgagca 22020agagctcagg gccatcgtca gagacctggg atgcgggccc tattttttgg
gcaccttcga 22080caagcgcttc cctggctttg tctccccaca caagctggcc tgcgccatcg
tcaacacggc 22140cggccgcgag accgggggcg tgcactggct ggcctttgcc tggaacccgc
gctccaaaac 22200atgcttcctc tttgacccct tcggcttttc ggaccagcgg ctcaagcaaa
tctacgagtt 22260cgagtacgag ggcttgctgc gtcgcagcgc catcgcctcc tcgcccgacc
gctgcgtcac 22320cctcgaaaag tccacccaga ccgtgcaggg gcccgactcg gccgcctgcg
gtctcttctg 22380ctgcatgttt ctgcacgcct ttgtgcactg gcctcagagt cccatggacc
gcaaccccac 22440catgaacttg ctgacggggg tgcccaactc catgctccaa agcccccagg
tcgagcccac 22500cctgcgccgc aaccaggagc agctctacag cttcctggag cgccactcgc
cctacttccg 22560ccgccacagc gcacagatca ggagggccac ctccttctgc cacttgcaag
agatgcaaga 22620agggtaataa cgatgtacac acttttttct caataaatgg catttttttt
ttatttatac 22680aagctctctg gggtattcat ttcccaccac caccacccgc cgttgtcgcc
atctggctct 22740atttagaaat cgaaagggtt ctgccgggag tcgccgtgcg ccacgggcag
ggacacgttg 22800cgatactggt agcgggtgcc ccacttgaac tcgggcacca ccaggcgagg
cagctcgggg 22860aagttttcgc tccacaggct gcgggtcagc accagcgcgt tcatcaggtc
gggcgccgag 22920atcttgaagt cgcagttggg gccgccgccc tgcgcgcgcg agttgcggta
caccgggttg 22980cagcactgga acaccaacag cgccgggtgc ttcacgctgg ccagcacgct
gcggtcggag 23040atcagctcgg cgtccaggtc ctccgcgttg ctcagcgcga acggggtcat
cttgggcact 23100tgccgcccca ggaagggcgc gtgccccggt ttcgagttgc agtcgcagcg
cagcgggatc 23160agcaggtgcc cgtgcccgga ctcggcgttg gggtacagcg cgcgcatgaa
ggcctgcatc 23220tggcggaagg ccatctgggc cttggcgccc tccgagaaga acatgccgca
ggacttgccc 23280gagaactggt ttgcggggca gctggcgtcg tgcaggcagc agcgcgcgtc
ggtgttggcg 23340atctgcacca cgttgcgccc ccaccggttc ttcacgatct tggccttgga
cgattgctcc 23400ttcagcgcgc gctgcccgtt ctcgctggtc acatccatct cgatcacatg
ttccttgttc 23460accatgctgc tgccgtgcag acacttcagc tcgccctccg tctcggtgca
gcggtgctgc 23520cacagcgcgc agcccgtggg ctcgaaagac ttgtaggtca cctccgcgaa
ggactgcagg 23580tacccctgca aaaagcggcc catcatggtc acgaaggtct tgttgctgct
gaaggtcagc 23640tgcagcccgc ggtgctcctc gttcagccag gtcttgcaca cggccgccag
cgcctccacc 23700tggtcgggca gcatcttgaa gttcaccttc agctcattct ccacgtggta
cttgtccatc 23760agcgtgcgcg ccgcctccat gcccttctcc caggccgaca ccagcggcag
gctcacgggg 23820ttcttcacca tcaccgtggc cgccgcctcc gccgcgcttt cgctttccgc
cccgctgttc 23880tcttcctctt cctcctcttc ctcgccgccg cccactcgca gcccccgcac
cacggggtcg 23940tcttcctgca ggcgctgcac cttgcgcttg ccgttgcgcc cctgcttgat
gcgcacgggc 24000gggttgctga agcccaccat caccagcgcg gcctcttctt gctcgtcctc
gctgtccaga 24060atgacctccg gggagggggg gttggtcatc ctcagtaccg aggcacgctt
ctttttcttc 24120ctgggggcgt tcgccagctc cgcggctgcg gccgctgccg aggtcgaagg
ccgagggctg 24180ggcgtgcgcg gcaccagcgc gtcttgcgag ccgtcctcgt cctcctcgga
ctcgagacgg 24240aggcgggccc gcttcttcgg gggcgcgcgg ggcggcggag gcggcggcgg
cgacggagac 24300ggggacgaga catcgtccag ggtgggtgga cggcgggccg cgccgcgtcc
gcgctcgggg 24360gtggtttcgc gctggtcctc ttcccgactg gccatctccc actgctcctt
ctcctatagg 24420cagaaagaga tcatggagtc tctcatgcga gtcgagaagg aggaggacag
cctaaccgcc 24480ccctctgagc cctccaccac cgccgccacc accgccaatg ccgccgcgga
cgacgcgccc 24540accgagacca ccgccagtac caccctcccc agcgacgcac ccccgctcga
gaatgaagtg 24600ctgatcgagc aggacccggg ttttgtgagc ggagaggagg atgaggtgga
tgagaaggag 24660aaggaggagg tcgccgcctc agtgccaaaa gaggataaaa agcaagacca
ggacgacgca 24720gataaggatg agacagcagt cgggcggggg aacggaagcc atgatgctga
tgacggctac 24780ctagacgtgg gagacgacgt gctgcttaag cacctgcacc gccagtgcgt
catcgtctgc 24840gacgcgctgc aggagcgctg cgaagtgccc ctggacgtgg cggaggtcag
ccgcgcctac 24900gagcggcacc tcttcgcgcc gcacgtgccc cccaagcgcc gggagaacgg
cacctgcgag 24960cccaacccgc gtctcaactt ctacccggtc ttcgcggtac ccgaggtgct
ggccacctac 25020cacatcttct tccaaaactg caagatcccc ctctcctgcc gcgctaaccg
cacccgcgcc 25080gacaaaaccc tgaccctgcg gcagggcgcc cacatacctg atattgcctc
tctggaggaa 25140gtgcccaaga tcttcgaggg tctcggtcgc gacgagaaac gggcggcgaa
cgctctgcac 25200ggagacagcg aaaacgagag tcactcgggg gtgctggtgg agctcgaggg
cgacaacgcg 25260cgcctggccg tactcaagcg cagcatagag gtcacccact ttgcctaccc
ggcgctcaac 25320ctgcccccca aggtcatgag tgtggtcatg ggcgagctca tcatgcgccg
cgctcagccc 25380ctggccgcgg atgcaaactt gcaagagtcc tccgaggaag gcctgcccgc
ggtcagcgac 25440gagcagctag cgcgctggct ggagacccgc gaccccgcgc agctggagga
gcggcgcaag 25500ctcatgatgg ccgcggtgct ggtcaccgtg gagctcgagt gtctgcagcg
cttcttcgcg 25560gaccccgaga tgcagcgcaa gctcgaggag accctgcact acaccttccg
ccagggctac 25620gtgcgccagg cctgcaagat ctccaacgtg gagctctgca acctggtctc
ctacctgggc 25680atcctgcacg agaaccgcct cgggcagaac gtcctgcact ccaccctcaa
aggggaggcg 25740cgccgcgact acatccgcga ctgcgcctac ctcttcctct gctacacctg
gcagacggcc 25800atgggggtct ggcagcagtg cctggaggag cgcaacctca aggagctgga
aaagctactc 25860aagcgcaccc tcagggacct ctggacgggc ttcaacgagc gctcggtggc
cgccgcgctg 25920gcggacatca tcttccccga gcgcctgctc aagaccctgc agcagggcct
gcccgacttc 25980accagccaga gcatgctgca gaactttagg actttcatcc tggagcgctc
gggcatcctg 26040cctgccactt gctgcgcgct gcccagcgac ttcgtgccca tcaagtacag
ggagtgcccg 26100ccgccgctct ggggccactg ctacctcttc cagctggcca actacctcgc
ctaccactcg 26160gacctcatgg aagacgtgag cggcgagggc ctgctcgagt gccactgccg
ctgcaacctc 26220tgcacgcccc accgctctct agtctgcaac ccgcagctgc tcagcgagag
tcagattatc 26280ggtaccttcg agctgcaggg tccctcgcct gacgagaagt ccgcggctcc
ggggctgaaa 26340ctcactccgg ggctgtggac ttccgcctac ctacgcaaat ttgtacctga
ggactaccac 26400gcccacgaga tcaggttcta cgaagaccaa tcccgcccgc ccaaggcgga
gctcaccgcc 26460tgcgtcatca cccaggggca catcctgggc caattgcaag ccatcaacaa
agcccgccga 26520gagttcttgc tgaaaaaggg tcggggggtg tacctggacc cccagtccgg
cgaggagcta 26580aacccgctac ccccgccgcc gccccagcag cgggaccttg cttcccagga
tggcacccag 26640aaagaagcag cagccgccgc cgccgcagcc atacatgctt ctggaggaag
aggaggagga 26700ctgggacagt caggcagagg aggtttcgga cgaggagcag gaggagatga
tggaagactg 26760ggaggaggac agcagcctag acgaggaagc ttcagaggcc gaagaggtgg
cagacgcaac 26820accatcaccc tcggtcgcag ccccctcgcc ggggcccctg aaatcctccg
aacccagcac 26880cagcgctata acctccgctc ctccggcgcc ggcgccaccc gcccgcagac
ccaaccgtag 26940atgggacacc acaggaaccg gggtcggtaa gtccaagtgc ccgccgccgc
caccgcagca 27000gcagcagcag cgccagggct accgctcgtg gcgcgggcac aagaacgcca
tagtcgcctg 27060cttgcaagac tgcgggggca acatctcttt cgcccggcgc ttcctgctat
tccaccacgg 27120ggtcgccttt ccccgcaatg tcctgcatta ctaccgtcat ctctacagcc
cctactgcag 27180cggcgaccca gaggcggcag cggcagccac agcggcgacc accacctagg
aagatatcct 27240ccgcgggcaa gacagcggca gcagcggcca ggagacccgc ggcagcagcg
gcgggagcgg 27300tgggcgcact gcgcctctcg cccaacgaac ccctctcgac ccgggagctc
agacacagga 27360tcttccccac tttgtatgcc atcttccaac agagcagagg ccaggagcag
gagctgaaaa 27420taaaaaacag atctctgcgc tccctcaccc gcagctgtct gtatcacaaa
agcgaagatc 27480agcttcggcg cacgctggag gacgcggagg cactcttcag caaatactgc
gcgctcactc 27540ttaaagacta gctccgcgcc cttctcgaat ttaggcggga gaaaactacg
tcatcgccgg 27600ccgccgccca gcccgcccag ccgagatgag caaagagatt cccacgccat
acatgtggag 27660ctaccagccg cagatgggac tcgcggcggg agcggcccag gactactcca
cccgcatgaa 27720ctacatgagc gcgggacccc acatgatctc acaggtcaac gggatccgcg
cccagcgaaa 27780ccaaatactg ctggaacagg cggccatcac cgccacgccc cgccataatc
tcaacccccg 27840aaattggccc gccgccctcg tgtaccagga aaccccctcc gccaccaccg
tactacttcc 27900gcgtgacgcc caggccgaag tccagatgac taactcaggg gcgcagctcg
cgggcggctt 27960tcgtcacggg gcgcggccgc tccgaccagg tataagacac ctgatgatca
gaggccgagg 28020tatccagctc aacgacgagt cggtgagctc ttcgctcggt ctccgtccgg
acggaacttt 28080ccagctcgcc ggatccggcc gctcttcgtt cacgccccgc caggcgtacc
tgactctgca 28140gacctcgtcc tcggagcccc gctccggagg catcggaacc ctccagttcg
tggaggagtt 28200cgtgccctcg gtctacttca accccttctc gggacctccc ggacgctacc
ccgaccagtt 28260cattccgaac tttgacgcgg tgaaggactc ggcggacggc tacgactgaa
tgtcaggtgc 28320cgaggcagag cagcttcgcc tgagacacct cgagcactgc cgccgccaca
agtgcttcgc 28380ccgcggttcc ggtgagttct gctactttca gctacccgag gagcataccg
aggggccggc 28440gcacggcgtc cgcctgacca cccagggcga ggttacctgt tccctcatcc
gggagttcac 28500cctccgtccc ctgctagtgg agcgggagcg gggtccctgt gtcctaacta
tcgcctgcaa 28560ctgccctaac cctggattac atcaagatct ttgctgtcat ctctgtgctg
agtttaataa 28620acgctgagat cagaatctac tggggctcct gtcgccatcc tgtgaacgcc
accgtcttca 28680cccaccccga ccaggcccag gcgaacctca cctgcggtct gcatcggagg
gccaagaagt 28740acctcacctg gtacttcaac ggcaccccct ttgtggttta caacagcttc
gacggggacg 28800gagtctccct gaaagaccag ctctccggtc tcagctactc catccacaag
aacaccaccc 28860tccaactctt ccctccctac ctgccgggaa cctacgagtg cgtcaccggc
cgctgcaccc 28920acctcacccg cctgatcgta aaccagagct ttccgggaac agataactcc
ctcttcccca 28980gaacaggagg tgagctcagg aaactccccg gggaccaggg cggagacgta
ccttcgaccc 29040ttgtggggtt aggatttttt attaccgggt tgctggctct tttaatcaaa
gcttccttga 29100gatttgttct ttccttctac gtgtatgaac acctcagcct ccaataactc
taccctttct 29160tcggaatcag gtgacttctc tgaaatcggg cttggtgtgc tgcttactct
gttgattttt 29220ttccttatca tactcagcct tctgtgcctc aggctcgccg cctgctgcgc
acacatctat 29280atctactgct ggttgctcaa gtgcaggggt cgccacccaa gatgaacagg
tacatggtcc 29340tatcgatcct aggcctgctg gccctggcgg cctgcagcgc cgccaaaaaa
gagattacct 29400ttgaggagcc cgcttgcaat gtaactttca agcccgaggg tgaccaatgc
accaccctcg 29460tcaaatgcgt taccaatcat gagaggctgc gcatcgacta caaaaacaaa
actggccagt 29520ttgcggtcta tagtgtgttt acgcccggag acccctctaa ctactctgtc
accgtcttcc 29580agggcggaca gtctaagata ttcaattaca ctttcccttt ttatgagtta
tgcgatgcgg 29640tcatgtacat gtcaaaacag tacaacctgt ggcctccctc tccccaggcg
tgtgtggaaa 29700atactgggtc ttactgctgt atggctttgg caatcactac gctcgctcta
atctgcacgg 29760tgctatacat aaaattcagg cagaggcgaa tctttatcga tgaaaagaaa
atgccttgat 29820cgctaacacc ggctttctat ctgcagaatg aatgcaatca cctccctact
aatcaccacc 29880accctccttg cgattgccca tgggttgaca cgaatcgaag tgccagtggg
gtccaatgtc 29940accatggtgg gccccgccgg caattccacc ctcatgtggg aaaaatttgt
ccgcaatcaa 30000tgggttcatt tctgctctaa ccgaatcagt atcaagccca gagccatctg
cgatgggcaa 30060aatctaactc tgatcaatgt gcaaatgatg gatgctgggt actattacgg
gcagcgggga 30120gaaatcatta attactggcg accccacaag gactacatgc tgcatgtagt
cgaggcactt 30180cccactacca cccccactac cacctctccc accaccacta ccaccactac
tactactact 30240actaccacta ccgctgcccg ccatacccgc aaaagcacca tgattagcac
aaagccccct 30300cgtgctcact cccacgccgg cgggcccatc ggtgcgacct cagaaaccac
cgagctttgc 30360ttctgccaat gcactaacgc cagcgctcat gaactgttcg acctggagaa
tgaggatgcc 30420cagcagagct ccgcttgcct gacccaggag gctgtggagc ccgttgccct
gaagcagatc 30480ggtgattcaa taattgactc ttcttctttt gccactcccg aataccctcc
cgattctact 30540ttccacatca cgggtaccaa agaccctaac ctctctttct acctgatgct
gctgctctgt 30600atctctgtgg tctcttccgc gctgatgtta ctggggatgt tctgctgcct
gatctgccgc 30660agaaagagaa aagctcgctc tcagggccaa ccactgatgc ccttccccta
ccccccggat 30720tttgcagata acaagatatg agctcgctgc tgacactaac cgctttacta
gcctgcgctc 30780taacccttgt cgcttgcgac tcgagattcc acaatgtcac agctgtggca
ggagaaaatg 30840ttactttcaa ctccacggcc gatacccagt ggtcgtggag tggctcaggt
agctacttaa 30900ctatctgcaa tagctccact tcccccagca tatccccaac caagtaccaa
tgcaatgcca 30960gcctgttcac cctcatcaac gcttccaccc tggacaatgg actctatgta
ggctatgtac 31020cctttggtgg gcaaggaaag acccacgctt acaacctgga agttcgccag
cccagaacca 31080ctacccaagc ttctcccacc accaccacca ccaccaccac caccatcacc
agcagcagca 31140gcagccacag cagcagcagc agattattga ctttggtttt ggccagctca
tctgccgcta 31200cccaggccat ctacagctct gtgcccgaaa ccactcagat ccaccgccca
gaaacgacca 31260ccgccaccac cctacacacc tccagcgatc agatgccgac caacatcacc
cccttggctc 31320ttcaaatggg acttacaagc cccactccaa aaccagtgga tgcggccgag
gtctccgccc 31380tcgtcaatga ctgggcgggg ctgggaatgt ggtggttcgc cataggcatg
atggcgctct 31440gcctgcttct gctctggctc atctgctgcc tccaccgcag gcgagccaga
ccccccatct 31500atagacccat cattgtcctg aaccccgata atgatgggat ccatagattg
gatggcctga 31560aaaacctact tttttctttt acagtatgat aaattgagac atgcctcgca
ttttcttgta 31620catgttcctt ctcccacctt ttctggggtg ttctacgctg gccgctgtgt
ctcacctgga 31680ggtagactgc ctctcaccct tcactgtcta cctgctttac ggattggtca
ccctcactct 31740catctgcagc ctaatcacag taatcatcgc cttcatccag tgcattgatt
acatctgtgt 31800gcgcctcgca tacttcagac accacccgca gtaccgagac aggaacattg
cccaacttct 31860aagactgctc taatcatgca taagactgtg atctgccttc tgatcctctg
catcctgccc 31920accctcacct cctgccagta caccacaaaa tctccgcgca aaagacatgc
ctcctgccgc 31980ttcacccaac tgtggaatat acccaaatgc tacaacgaaa agagcgagct
ctccgaagct 32040tggctgtatg gggtcatctg tgtcttagtt ttctgcagca ctgtctttgc
cctcatgatc 32100tacccctact ttgatttggg atggaacgcg atcgatgcca tgaattaccc
cacctttccc 32160gcacccgaga taattccact gcgacaagtt gtacccgttg tcgttaatca
acgcccccca 32220tcccctacgc ccactgaaat cagctacttt aacctaacag gcggagatga
ctgacgccct 32280agatctagaa atggacggca tcagtaccga gcagcgtctc ctagagaggc
gcaggcaggc 32340ggctgagcaa gagcgcctca atcaggagct ccgagatctc gttaacctgc
accagtgcaa 32400aagaggcatc ttttgtctgg taaagcaggc caaagtcacc tacgagaaga
ccggcaacag 32460ccaccgcctc agttacaaat tgcccaccca gcgccagaag ctggtgctca
tggtgggtga 32520gaatcccatc accgtcaccc agcactcggt agagaccgag gggtgtctgc
actctccctg 32580tcggggtcca gaagacctct gcaccctggt aaagaccctg tgcggtctca
gagatttagt 32640cccctttaac taatcaaaca ctggaatcaa taaaaagaat cacttactta
aaatcagaca 32700gcaggtctct gtccagttta ttcagcagca cctccttccc ctcctcccaa
ctctggtact 32760ccaaacgcct tctggcggca aacttcctcc acaccctgaa gggaatgtca
gattcttgct 32820cctgtccctc cgcacccact atcttcatgt tgttgcagat gaagcgcacc
aaaacgtctg 32880acgagagctt caaccccgtg tacccctatg acacggaaag cggccctccc
tccgtccctt 32940tcctcacccc tcccttcgtg tctcccgatg gattccaaga aagccccccc
ggggtcctgt 33000ctctgaacct ggccgagccc ctggtcactt cccacggcat gctcgccctg
aaaatgggaa 33060gtggcctctc cctggacgac gctggcaacc tcacctctca agatatcacc
accgctagcc 33120ctcccctcaa aaaaaccaag accaacctca gcctagaaac ctcatccccc
ctaactgtaa 33180gcacctcagg cgccctcacc gtagcagccg ccgctcccct ggcagtggcc
ggcacctccc 33240tcaccatgca atcagaggcc cccctgacag tacaggatgc aaaactcacc
ctggccacca 33300aaggccccct gaccgtgtct gaaggcaaac tggccttgca aacatcggcc
ccgctgacgg 33360ccgctgacag cagcaccctc accgttagcg ccacaccacc aattaatgta
agcagtggaa 33420gtttaggctt agacatggaa gaccctatgt atactcacga tggaaaactg
ggaataagaa 33480ttgggggtcc actaagagta gtagacagct tgcacacact cactgtagtt
accggaaatg 33540gactaactgt agataacaat gccctccaaa ctagagttac gggcgcccta
ggttatgaca 33600catcaggaaa tctacaattg agagctgcag gaggtatgcg aattgatgca
aatggccaac 33660ttatccttaa tgtggcatac ccatttgatg ctcagaacaa tctcagcctt
agacttggtc 33720agggacccct gtatataaac acagaccaca acctggattt gaattgcaac
agaggtctaa 33780ccacaactac caccaacaac acaaaaaaac ttgagactaa aattagctca
ggcttagact 33840atgacaccaa tggtgctgtc attattaaac ttggcactgg tctaagcttc
gacaacacag 33900gcgccctaac tgtgggaaac actggtgatg ataaactgac tctgtggacg
accccagacc 33960catctccaaa ttgcagaatt cactcagaca aagactgcaa gtttactcta
gtcctaacta 34020agtgtggaag ccaaatcctg gcctctgtcg ccgccctagc ggtatcagga
aatctggctt 34080cgataacagg caccgttgcc agcgttacca tctttctcag atttgatcag
aatggagtgc 34140ttatggaaaa ctcctcgcta gacaggcagt actggaactt cagaaatggc
aactcaacta 34200acgctgcccc ctacaccaat gcagttgggt tcatgccaaa cctcgcagca
taccccaaaa 34260cgcaaagcca gactgctaaa aacaacattg taagtcaggt ttacttgaat
ggagacaaat 34320ccaaacccat gacccttacc atcaccctca atggaactaa tgaatccagt
gaaactagcc 34380aggtgagtca ctactccatg tcatttacat gggcttggga aagtgggcaa
tatgccactg 34440aaacctttgc caccaactcc ttcacctttt cttacattgc tgaacaataa
aaagcatgac 34500actgatgttc atttctgatt cttattttat tattttcaaa cacaacaaaa
tcattcaagt 34560cattcttcca tcttagctta atagacacag tagcttaata gacccagtag
tgcaaagccc 34620cattctagct tatagatcag acagtgataa ttaaccacca ccaccaccat
accttttgat 34680tcaggaaatc atgatcatca caggatccta gtcttcaggc cgccccctcc
ctcccaagac 34740acagaataca cagtcctctc cccccgactg gctttaaata acaccatctg
gttggtcaca 34800gacatgttct taggggttat attccacacg gtctcctgcc gcgccaggcg
ctcgtcggtg 34860atgttgataa actctcccgg cagctcgctc aagttcacgt cgctgtccag
cggctgaacc 34920tccggctgac gcgataactg tgcgaccggc tgctggacaa acggaggccg
cgcctacaag 34980ggggtagagt cataatcctc ggtcaggata gggcggtgat gcagcagcag
cgagcgaaac 35040atctgctgcc gccgccgctc cgtccggcag gaaaacaaca agccggtggt
ctcctccgcg 35100ataatccgca ccgcccgcag catcagcttc ctcgttctcc gcgcgcagca
cctcaccctg 35160atctcgctca agtcggcgca gtaggtacag cacagcacca cgatgttatt
catgatccca 35220cagtgcaggg cgctgtatcc aaagctcatg ccgggaacca ccgcccccac
gtggccatcg 35280taccacaagc gcacgtaaat taagtgtcga cccctcatga acgtgctgga
cacaaacatt 35340acttccttgg gcatgttgta attcaccacc tcccggtacc agataaacct
ctggttaaac 35400agggcacctt ccaccaccat cctgaaccaa gaggccagaa cctgcccacc
ggctatgcac 35460tgcagggaac ccgggttgga acaatgacaa tgcagactcc aaggctcgta
accgtggatc 35520atccggctgc tgaaggcatc gatgttggca caacacagac acacgtgcat
gcactttctc 35580atgattagca gctcttccct cgtcaggatc atatcccaag gaataaccca
ttcttgaatc 35640aacgtaaaac ccacacagca gggaaggcct cgcacataac tcacgttgtg
catggtcagc 35700gtgttgcatt ctggaaacag cggatgatcc tccagtatcg aggcgcgggt
ctccttctca 35760cagggaggta aagggtccct gctgtacgga ctgcgccggg acgaccgaga
tcgtgttgag 35820cgtagtgtca tggaaaaggg aacgccggac gtggtcatac ttcttgaagc
agaaccaggt 35880tcgcgcgtgg caggcctcct tgcgtctgcg gtctcgccgt ctagctcgct
ccgtgtgata 35940gttgtagtac agccactccc gcagagcgtc gaggcgcacc ctggcttccg
gatctatgta 36000gactccgtct tgcaccgcgg ccctgataat atccaccacc gtagaataag
caacacccag 36060ccaagcaata cactcgctct gcgagcggca gacaggagga gcgggcagag
atgggagaac 36120catgataaaa aacttttttt aaagaatatt ttccaattct tcgaaagtaa
gatctatcaa 36180gtggcagcgc tcccctccac tggcgcggtc aaactctacg gccaaagcac
agacaacggc 36240atttctaaga tgttccttaa tggcgtccaa aagacacacc gctctcaagt
tgcagtaaac 36300tatgaatgaa aacccatccg gctgattttc caatatagac gcgccggcgg
cgtccaccaa 36360acccagataa ttttcttctc tccagcggtt tagaatctgt ctaagcaaat
cccttatatc 36420aagtccggcc atgccaaaaa tctgctcaag agcgccctcc accttcatga
ccaagcagcg 36480catcatgatt gcaaaaattc aggttcttca gagacctgta taagattcaa
aatgggaaca 36540ttaacaaaaa ttcctctgtc gcgcagatcc cttcgcaggg caagctgaac
ataatcagac 36600aggtctgaac ggaccagtga ggccaaatcc ccaccaggaa ccagatccag
agaccctata 36660ctgattatga cgcgcatact cggggctatg ctgaccagcg tagcgccgat
gtaggcgtgc 36720tgcatgggcg gcgagataaa atgcaaagtg ctggttaaaa aatcaggcaa
agcctcgcgc 36780aaaaaagcta acacatcata atcatgctca tgcaggtagt tgcaggtaag
ctcaggaacc 36840aaaacggaat aacacacgat tttcctctca aacatgactt cgcggatact
gcgtaaaaca 36900aaaattataa ataaaaaatt aattaactta aacattggaa gcctgtctca
caacaggaaa 36960aaccacttta atcaacataa gacgggccac gggcatgccg gcatagccgt
aaaaaaattg 37020gtccccgtga ttaacaagta ccacagacag ctccccggtc atgtcggggg
tcatcatgtg 37080agactctgta tacacgtctg gattgtgaac atcagacaaa caaagaaatc
gagccacgta 37140gcccggaggt ataatcaccc gcaggcggag gtacagcaaa acgaccccca
taggaggaat 37200cacaaaatta gtaggagaaa aaaatacata aacaccagaa aaaccctgtt
gctgaggcaa 37260aatagcgccc tcccgatcca aaacaacata aagcgcttcc acaggagcag
ccataacaaa 37320gacccgagtc ttaccagtaa aagaaaaaag atctctcaac gcagcaccag
caccaacact 37380tcgcagtgta aaaggccaag tgccgagaga gtatatatag gaataaaaag
tgacgtaaac 37440gggcaaagtc caaaaaacgc ccagaaaaac cgcacgcgaa cctacgcccc
gaaacgaaag 37500ccaaaaaaca ctagacactc ccttccggcg tcaacttccg ctttcccacg
ctacgtcact 37560tgccccagtc aaacaaacta catatcccga acttccaagt cgccacgccc
aaaacaccgc 37620ctacacctcc ccgcccgccg gcccgccccc aaacccgcct cccgccccgc
gccccgcctc 37680gcgccgccca tctcattatc atattggctt caatccaaaa taaggtatat
tattgatgat 37740g
3774110593PRTPan troglodytes 10Met Arg Arg Ala Ala Met Tyr Gln
Glu Gly Pro Pro Pro Ser Tyr Glu1 5 10
15Ser Val Val Gly Ala Ala Ala Ala Ala Pro Ser Ser Pro Phe
Ala Ser 20 25 30Gln Leu Leu
Glu Pro Pro Tyr Val Pro Pro Arg Tyr Leu Arg Pro Thr 35
40 45Gly Gly Arg Asn Ser Ile Arg Tyr Ser Glu Leu
Ala Pro Leu Phe Asp 50 55 60Thr Thr
Arg Val Tyr Leu Val Asp Asn Lys Ser Ala Asp Val Ala Ser65
70 75 80Leu Asn Tyr Gln Asn Asp His
Ser Asn Phe Leu Thr Thr Val Ile Gln 85 90
95Asn Asn Asp Tyr Ser Pro Ser Glu Ala Ser Thr Gln Thr
Ile Asn Leu 100 105 110Asp Asp
Arg Ser His Trp Gly Gly Asp Leu Lys Thr Ile Leu His Thr 115
120 125Asn Met Pro Asn Val Asn Glu Phe Met Phe
Thr Asn Lys Phe Lys Ala 130 135 140Arg
Val Met Val Ser Arg Ser His Thr Lys Glu Asp Arg Val Glu Leu145
150 155 160Lys Tyr Glu Trp Val Glu
Phe Glu Leu Pro Glu Gly Asn Tyr Ser Glu 165
170 175Thr Met Thr Ile Asp Leu Met Asn Asn Ala Ile Val
Glu His Tyr Leu 180 185 190Lys
Val Gly Arg Gln Asn Gly Val Leu Glu Ser Asp Ile Gly Val Lys 195
200 205Phe Asp Thr Arg Asn Phe Arg Leu Gly
Leu Asp Pro Val Thr Gly Leu 210 215
220Val Met Pro Gly Val Tyr Thr Asn Glu Ala Phe His Pro Asp Ile Ile225
230 235 240Leu Leu Pro Gly
Cys Gly Val Asp Phe Thr Tyr Ser Arg Leu Ser Asn 245
250 255Leu Leu Gly Ile Arg Lys Arg Gln Pro Phe
Gln Glu Gly Phe Arg Ile 260 265
270Thr Tyr Glu Asp Leu Glu Gly Gly Asn Ile Pro Ala Leu Leu Asp Val
275 280 285Glu Ala Tyr Gln Asp Ser Leu
Lys Glu Asn Glu Ala Gly Gln Glu Asp 290 295
300Thr Thr Pro Ala Ala Ser Ala Ala Ala Glu Gln Gly Glu Asp Ala
Ala305 310 315 320Asp Thr
Ala Ala Ala Asp Gly Ala Glu Ala Asp Pro Ala Met Val Val
325 330 335Glu Ala Pro Glu Gln Glu Glu
Asp Met Asn Asp Ser Ala Val Arg Gly 340 345
350Asp Thr Phe Val Thr Arg Gly Glu Glu Lys Gln Ala Glu Ala
Glu Ala 355 360 365Ala Ala Glu Glu
Lys Gln Leu Ala Ala Ala Ala Ala Ala Ala Ala Leu 370
375 380Ala Ala Ala Glu Ala Glu Ser Glu Gly Thr Lys Pro
Ala Lys Glu Pro385 390 395
400Val Ile Lys Pro Leu Thr Glu Asp Ser Lys Lys Arg Ser Tyr Asn Leu
405 410 415Leu Lys Asp Ser Thr
Asn Thr Ala Tyr Arg Ser Trp Tyr Leu Ala Tyr 420
425 430Asn Tyr Gly Asp Pro Ser Thr Gly Val Arg Ser Trp
Thr Leu Leu Cys 435 440 445Thr Pro
Asp Val Thr Cys Gly Ser Glu Gln Val Tyr Trp Ser Leu Pro 450
455 460Asp Met Met Gln Asp Pro Val Thr Phe Arg Ser
Thr Arg Gln Val Ser465 470 475
480Asn Phe Pro Val Val Gly Ala Glu Leu Leu Pro Val His Ser Lys Ser
485 490 495Phe Tyr Asn Asp
Gln Ala Val Tyr Ser Gln Leu Ile Arg Gln Phe Thr 500
505 510Ser Leu Thr His Val Phe Asn Arg Phe Pro Glu
Asn Gln Ile Leu Ala 515 520 525Arg
Pro Pro Ala Pro Thr Ile Thr Thr Val Ser Glu Asn Val Pro Ala 530
535 540Leu Thr Asp His Gly Thr Leu Pro Leu Arg
Asn Ser Ile Gly Gly Val545 550 555
560Gln Arg Val Thr Val Thr Asp Ala Arg Arg Arg Thr Cys Pro Tyr
Val 565 570 575Tyr Lys Ala
Leu Gly Ile Val Ser Pro Arg Val Leu Ser Ser Arg Thr 580
585 590Phe11960PRTPan troglodytes 11Met Ala Thr
Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser1 5
10 15Gly Gln Asp Ala Ser Glu Tyr Leu Ser
Pro Gly Leu Val Gln Phe Ala 20 25
30Arg Ala Thr Glu Ser Tyr Phe Ser Leu Ser Asn Lys Phe Arg Asn Pro
35 40 45Thr Val Ala Pro Thr His Asp
Val Thr Thr Asp Arg Ser Gln Arg Leu 50 55
60Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr65
70 75 80Lys Ala Arg Phe
Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met 85
90 95Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val
Leu Asp Arg Gly Pro Thr 100 105
110Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ser Leu Ala Pro Lys Gly
115 120 125Ala Pro Asn Ser Cys Glu Trp
Glu Gln Glu Glu Thr Gln Ala Val Glu 130 135
140Glu Ala Ala Glu Glu Glu Glu Glu Asp Ala Asp Gly Gln Ala Glu
Glu145 150 155 160Glu Gln
Ala Ala Thr Lys Lys Thr His Val Tyr Ala Gln Ala Pro Leu
165 170 175Ser Gly Glu Lys Ile Ser Lys
Asp Gly Leu Gln Ile Gly Thr Asp Ala 180 185
190Thr Ala Thr Glu Gln Lys Pro Ile Tyr Ala Asp Pro Thr Phe
Gln Pro 195 200 205Glu Pro Gln Ile
Gly Glu Ser Gln Trp Asn Glu Ala Asp Ala Thr Val 210
215 220Ala Gly Gly Arg Val Leu Lys Lys Ser Thr Pro Met
Lys Pro Cys Tyr225 230 235
240Gly Ser Tyr Ala Arg Pro Thr Asn Ala Asn Gly Gly Gln Gly Val Leu
245 250 255Thr Ala Asn Ala Gln
Gly Gln Leu Glu Ser Gln Val Glu Met Gln Phe 260
265 270Phe Ser Thr Ser Glu Asn Ala Arg Asn Glu Ala Asn
Asn Ile Gln Pro 275 280 285Lys Leu
Val Leu Tyr Ser Glu Asp Val His Met Glu Thr Pro Asp Thr 290
295 300His Leu Ser Tyr Lys Pro Ala Lys Ser Asp Asp
Asn Ser Lys Ile Met305 310 315
320Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile Gly Phe Arg
325 330 335Asp Asn Phe Ile
Gly Leu Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly 340
345 350Val Leu Ala Gly Gln Ala Ser Gln Leu Asn Ala
Val Val Asp Leu Gln 355 360 365Asp
Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp Ser Met Gly 370
375 380Asp Arg Thr Arg Tyr Phe Ser Met Trp Asn
Gln Ala Val Asp Ser Tyr385 390 395
400Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr Glu Asp Glu
Leu 405 410 415Pro Asn Tyr
Cys Phe Pro Leu Gly Gly Ile Gly Val Thr Asp Thr Tyr 420
425 430Gln Ala Val Lys Thr Asn Asn Gly Asn Asn
Gly Gly Gln Val Thr Trp 435 440
445Thr Lys Asp Glu Thr Phe Ala Asp Arg Asn Glu Ile Gly Val Gly Asn 450
455 460Asn Phe Ala Met Glu Ile Asn Leu
Ser Ala Asn Leu Trp Arg Asn Phe465 470
475 480Leu Tyr Ser Asn Val Ala Leu Tyr Leu Pro Asp Lys
Leu Lys Tyr Asn 485 490
495Pro Ser Asn Val Asp Ile Ser Asp Asn Pro Asn Thr Tyr Asp Tyr Met
500 505 510Asn Lys Arg Val Val Ala
Pro Gly Leu Val Asp Cys Tyr Ile Asn Leu 515 520
525Gly Ala Arg Trp Ser Leu Asp Tyr Met Asp Asn Val Asn Pro
Phe Asn 530 535 540His His Arg Asn Ala
Gly Leu Arg Tyr Arg Ser Met Leu Leu Gly Asn545 550
555 560Gly Arg Tyr Val Pro Phe His Ile Gln Val
Pro Gln Lys Phe Phe Ala 565 570
575Ile Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr Tyr Glu Trp Asn
580 585 590Phe Arg Lys Asp Val
Asn Met Val Leu Gln Ser Ser Leu Gly Asn Asp 595
600 605Leu Arg Val Asp Gly Ala Ser Ile Lys Phe Glu Ser
Ile Cys Leu Tyr 610 615 620Ala Thr Phe
Phe Pro Met Ala His Asn Thr Ala Ser Thr Leu Glu Ala625
630 635 640Met Leu Arg Asn Asp Thr Asn
Asp Gln Ser Phe Asn Asp Tyr Leu Ser 645
650 655Ala Ala Asn Met Leu Tyr Pro Ile Pro Ala Asn Ala
Thr Asn Val Pro 660 665 670Ile
Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly Trp Ala Phe 675
680 685Thr Arg Leu Lys Thr Lys Glu Thr Pro
Ser Leu Gly Ser Gly Phe Asp 690 695
700Pro Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu Asp Gly Thr Phe705
710 715 720Tyr Leu Asn His
Thr Phe Lys Lys Val Ser Val Thr Phe Asp Ser Ser 725
730 735Val Ser Trp Pro Gly Asn Asp Arg Leu Leu
Thr Pro Asn Glu Phe Glu 740 745
750Ile Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val Ala Gln Cys Asn
755 760 765Met Thr Lys Asp Trp Phe Leu
Val Gln Met Leu Ala Asn Tyr Asn Ile 770 775
780Gly Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr Lys Asp Arg Met
Tyr785 790 795 800Ser Phe
Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val Asp Gln
805 810 815Thr Lys Tyr Lys Asp Tyr Gln
Glu Val Gly Ile Ile His Gln His Asn 820 825
830Asn Ser Gly Phe Val Gly Tyr Leu Ala Pro Thr Met Arg Glu
Gly Gln 835 840 845Ala Tyr Pro Ala
Asn Phe Pro Tyr Pro Leu Ile Gly Lys Thr Ala Val 850
855 860Asp Ser Ile Thr Gln Lys Lys Phe Leu Cys Asp Arg
Thr Leu Trp Arg865 870 875
880Ile Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala Leu Ser Asp Leu
885 890 895Gly Gln Asn Leu Leu
Tyr Ala Asn Ser Ala His Ala Leu Asp Met Thr 900
905 910Phe Glu Val Asp Pro Met Asp Glu Pro Thr Leu Leu
Tyr Val Leu Phe 915 920 925Glu Val
Phe Asp Val Val Arg Val His Gln Pro His Arg Gly Val Ile 930
935 940Glu Thr Val Tyr Leu Arg Thr Pro Phe Ser Ala
Gly Asn Ala Thr Thr945 950 955
96012543PRTPan troglodytes 12Met Lys Arg Thr Lys Thr Ser Asp Glu Ser
Phe Asn Pro Val Tyr Pro1 5 10
15Tyr Asp Thr Glu Ser Gly Pro Pro Ser Val Pro Phe Leu Thr Pro Pro
20 25 30Phe Val Ser Pro Asp Gly
Phe Gln Glu Ser Pro Pro Gly Val Leu Ser 35 40
45Leu Asn Leu Ala Glu Pro Leu Val Thr Ser His Gly Met Leu
Ala Leu 50 55 60Lys Met Gly Ser Gly
Leu Ser Leu Asp Asp Ala Gly Asn Leu Thr Ser65 70
75 80Gln Asp Ile Thr Thr Ala Ser Pro Pro Leu
Lys Lys Thr Lys Thr Asn 85 90
95Leu Ser Leu Glu Thr Ser Ser Pro Leu Thr Val Ser Thr Ser Gly Ala
100 105 110Leu Thr Val Ala Ala
Ala Ala Pro Leu Ala Val Ala Gly Thr Ser Leu 115
120 125Thr Met Gln Ser Glu Ala Pro Leu Thr Val Gln Asp
Ala Lys Leu Thr 130 135 140Leu Ala Thr
Lys Gly Pro Leu Thr Val Ser Glu Gly Lys Leu Ala Leu145
150 155 160Gln Thr Ser Ala Pro Leu Thr
Ala Ala Asp Ser Ser Thr Leu Thr Val 165
170 175Ser Ala Thr Pro Pro Ile Asn Val Ser Ser Gly Ser
Leu Gly Leu Asp 180 185 190Met
Glu Asp Pro Met Tyr Thr His Asp Gly Lys Leu Gly Ile Arg Ile 195
200 205Gly Gly Pro Leu Arg Val Val Asp Ser
Leu His Thr Leu Thr Val Val 210 215
220Thr Gly Asn Gly Leu Thr Val Asp Asn Asn Ala Leu Gln Thr Arg Val225
230 235 240Thr Gly Ala Leu
Gly Tyr Asp Thr Ser Gly Asn Leu Gln Leu Arg Ala 245
250 255Ala Gly Gly Met Arg Ile Asp Ala Asn Gly
Gln Leu Ile Leu Asn Val 260 265
270Ala Tyr Pro Phe Asp Ala Gln Asn Asn Leu Ser Leu Arg Leu Gly Gln
275 280 285Gly Pro Leu Tyr Ile Asn Thr
Asp His Asn Leu Asp Leu Asn Cys Asn 290 295
300Arg Gly Leu Thr Thr Thr Thr Thr Asn Asn Thr Lys Lys Leu Glu
Thr305 310 315 320Lys Ile
Ser Ser Gly Leu Asp Tyr Asp Thr Asn Gly Ala Val Ile Ile
325 330 335Lys Leu Gly Thr Gly Leu Ser
Phe Asp Asn Thr Gly Ala Leu Thr Val 340 345
350Gly Asn Thr Gly Asp Asp Lys Leu Thr Leu Trp Thr Thr Pro
Asp Pro 355 360 365Ser Pro Asn Cys
Arg Ile His Ser Asp Lys Asp Cys Lys Phe Thr Leu 370
375 380Val Leu Thr Lys Cys Gly Ser Gln Ile Leu Ala Ser
Val Ala Ala Leu385 390 395
400Ala Val Ser Gly Asn Leu Ala Ser Ile Thr Gly Thr Val Ala Ser Val
405 410 415Thr Ile Phe Leu Arg
Phe Asp Gln Asn Gly Val Leu Met Glu Asn Ser 420
425 430Ser Leu Asp Arg Gln Tyr Trp Asn Phe Arg Asn Gly
Asn Ser Thr Asn 435 440 445Ala Ala
Pro Tyr Thr Asn Ala Val Gly Phe Met Pro Asn Leu Ala Ala 450
455 460Tyr Pro Lys Thr Gln Ser Gln Thr Ala Lys Asn
Asn Ile Val Ser Gln465 470 475
480Val Tyr Leu Asn Gly Asp Lys Ser Lys Pro Met Thr Leu Thr Ile Thr
485 490 495Leu Asn Gly Thr
Asn Glu Ser Ser Glu Thr Ser Gln Val Ser His Tyr 500
505 510Ser Met Ser Phe Thr Trp Ala Trp Glu Ser Gly
Gln Tyr Ala Thr Glu 515 520 525Thr
Phe Ala Thr Asn Ser Phe Thr Phe Ser Tyr Ile Ala Glu Gln 530
535 5401336182DNAArtificial SequenceM72-ChAd3
construct 13catcatcaat aatatacctt attttggatt gaagccaata tgataatgag
atgggcggcg 60cgaggcgggg cgcggggcgg gaggcgggtt tgggggcggg ccggcgggcg
gggcggtgtg 120gcggaagtgg actttgtaag tgtggcggat gtgacttgct agtgccgggc
gcggtaaaag 180tgacgttttc cgtgcgcgac aacgcccccg ggaagtgaca tttttcccgc
ggtttttacc 240ggatgttgta gtgaatttgg gcgtaaccaa gtaagatttg gccattttcg
cgggaaaact 300gaaacgggga agtgaaatct gattaatttt gcgttagtca taccgcgtaa
tatttgtcta 360gggccgaggg actttggccg attacgtgga ggactcgccc aggtgttttt
tgaggtgaat 420ttccgcgttc cgggtcaaag tctccgtttt attattatag gatatcccat
tgcatacgtt 480gtatccatat cataatatgt acatttatat tggctcatgt ccaacattac
cgccatgttg 540acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
ttcatagccc 600atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
gaccgcccaa 660cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
caatagggac 720tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
cagtacatca 780agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
ggcccgcctg 840gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
tctacgtatt 900agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
gtggatagcg 960gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
gtttgttttg 1020gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
tgacgcaaat 1080gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctcccta
tcagtgatag 1140agatctccct atcagtgata gagatcgtcg acgagctcgt ttagtgaacc
gtcagatcgc 1200ctggagacgc catccacgct gttttgacct ccatagaaga caccgggacc
gatccagcct 1260ccgcggccgg gaacggtgca ttggaacgcg gattccccgt gccaagagtg
agatcttccg 1320tttatctagg taccagatat cgccaccatg accgccgcca gcgacaactt
ccagctgtct 1380cagggcggcc agggcttcgc catccctatc ggccaagcta tggccattgc
tggacagatc 1440agaagcggcg gaggcagccc taccgtgcat atcggcccta ccgccttcct
gggcctgggc 1500gtggtggaca acaacggcaa cggcgccaga gtgcagcggg tggtcggatc
tgcccctgcc 1560gcaagcctgg gcatcagcac cggggatgtg atcaccgccg tggatggcgc
ccctatcaac 1620agcgccacag ccatggccga cgccctgaat ggacaccacc ccggcgacgt
gatcagcgtg 1680acctggcaga ccaagagcgg aggcaccaga accggcaacg tgacactggc
cgagggacct 1740cccgccgagt tcatggtgga tttcggcgcc ctgccccccg agatcaactc
cgccaggatg 1800tatgccggcc ctggcagcgc ctctctggtg gccgctgctc agatgtggga
cagcgtggcc 1860agcgatctgt tcagcgccgc ctccgccttc cagtccgtgg tctggggcct
gaccgtgggc 1920agctggatcg gaagcagtgc cggcctgatg gtggctgccg cctctcccta
cgtggcctgg 1980atgtcagtca cagccggcca ggccgaactg actgccgctc aagtgcgagt
ggctgctgct 2040gcctatgaga cagcctacgg cctgacagtg cccccacccg tgatcgccga
gaaccgggcc 2100gagctgatga tcctgatcgc caccaacctg ctgggccaga acacccccgc
cattgccgtg 2160aacgaggccg agtacggcga gatgtgggcc caggacgccg ctgccatgtt
tggctatgcc 2220gctgctacag ccaccgccac tgccaccctg ctgcccttcg aagaggcccc
cgagatgacc 2280tctgccggcg gactgctgga acaggccgct gccgtggaag aggccagcga
cacagccgcc 2340gctaaccagc tgatgaacaa cgtgccccag gccctgcagc agctggcaca
gcctacacag 2400ggcaccaccc cttctagcaa gctcggcggc ctgtggaaaa ccgtgtcccc
ccaccggtcc 2460cccatcagca acatggtgtc catggccaac aaccacatga gcatgaccaa
cagcggcgtg 2520tccatgacca ataccctgag cagcatgctg aagggctttg ccccagccgc
tgccgctcag 2580gctgtgcaga cagctgctca gaatggcgtg cgggccatga gcagcctggg
cagttccctg 2640ggcagctctg gactgggagg gggcgtggcc gccaatctgg gcagagccgc
tagcgtgggc 2700agcctgtctg tgcctcaagc ctgggctgct gccaatcagg ccgtgacacc
agccgctaga 2760gccctgcctc tgaccagcct gacctctgct gccgagaggg gccctggcca
gatgctggga 2820ggactgcctg tgggccagat gggagccaga gccggcggag gactgagcgg
cgtgctgaga 2880gtgcccccca gaccctacgt gatgccccac tctcccgccg ctggcgatat
tgcccctccc 2940gccctgagcc aggacagatt cgccgacttc cctgccctgc ccctggatcc
ttctgccatg 3000gtggctcaag tgggacccca ggtggtgaac atcaacacca agctgggcta
caacaacgcc 3060gtgggagccg gcaccggcat cgtgatcgac cccaatggcg tggtgctgac
caacaatcac 3120gtgatcgctg gcgccaccga catcaacgcc ttcagcgtgg gctccggcca
gacctacggc 3180gtggacgtgg tcggatacga ccggacccag gatgtggccg tgctgcagct
gagaggcgct 3240ggcggactgc cttctgccgc cattggaggc ggagtggccg tgggagaacc
tgtggtggcc 3300atgggcaata gcggcggaca gggcggcaca cctagagctg tgcctggaag
agtggtggcc 3360ctgggacaga ccgtgcaggc cagcgatagc ctgacaggcg ccgaggaaac
cctgaacggc 3420ctgatccagt tcgacgccgc catccagcct ggggatgctg gcggacctgt
ggtgaacgga 3480ctgggccagg tggtcggaat gaataccgcc gcctcctaat agtgagcggc
cgcgatctgc 3540tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
ccttgaccct 3600ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
cgcattgtct 3660gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
gggaggattg 3720ggaagacaat agcaggcatg ctggggatgc ggtgggctct agatatcagc
gatcgctgag 3780gtgggtgagt gggcgtggcc tggggtggtc atgaaaatat ataagttggg
ggtcttaggg 3840tctctttatt tgtgttgcag agaccgccgg agccatgagc gggagcagca
gcagcagcag 3900tagcagcagc gccttggatg gcagcatcgt gagcccttat ttgacgacgc
ggatgcccca 3960ctgggccggg gtgcgtcaga atgtgatggg ctccagcatc gacggccgac
ccgtcctgcc 4020cgcaaattcc gccacgctga cctatgcgac cgtcgcgggg acgccgttgg
acgccaccgc 4080cgccgccgcc gccaccgcag ccgcctcggc cgtgcgcagc ctggccacgg
actttgcatt 4140cctgggacca ctggcgacag gggctacttc tcgggccgct gctgccgccg
ttcgcgatga 4200caagctgacc gccctgctgg cgcagttgga tgcgcttact cgggaactgg
gtgacctttc 4260tcagcaggtc atggccctgc gccagcaggt ctcctccctg caagctggcg
ggaatgcttc 4320tcccacaaat gccgtttaag ataaataaaa ccagactctg tttggattaa
agaaaagtag 4380caagtgcatt gctctcttta tttcataatt ttccgcgcgc gataggccct
agaccagcgt 4440tctcggtcgt tgagggtgcg gtgtatcttc tccaggacgt ggtagaggtg
gctctggacg 4500ttgagataca tgggcatgag cccgtcccgg gggtggaggt agcaccactg
cagagcttca 4560tgctccgggg tggtgttgta gatgatccag tcgtagcagg agcgctgggc
atggtgccta 4620aaaatgtcct tcagcagcag gccgatggcc agggggaggc ccttggtgta
agtgtttaca 4680aaacggttaa gttgggaagg gtgcattcgg ggagagatga tgtgcatctt
ggactgtatt 4740tttagattgg cgatgtttcc gcccagatcc cttctgggat tcatgttgtg
caggaccacc 4800agtacagtgt atccggtgca cttggggaat ttgtcatgca gcttagaggg
aaaagcgtgg 4860aagaacttgg agacgccctt gtggcctccc agattttcca tgcattcgtc
catgatgatg 4920gcaatgggcc cgcgggaggc agcttgggca aagatatttc tggggtcgct
gacgtcgtag 4980ttgtgttcca gggtgaggtc gtcataggcc atttttacaa agcgcgggcg
gagggtgccc 5040gactggggga tgatggtccc ctctggccct ggggcgtagt tgccctcgca
gatctgcatt 5100tcccaggcct taatctcgga ggggggaatc atatccacct gcggggcgat
gaagaaaacg 5160gtttccggag ccggggagat taactgggat gagagcaggt ttctaagcag
ctgtgatttt 5220ccacaaccgg tgggcccata aataacacct ataaccggtt gcagctggta
gtttagagag 5280ctgcagctgc cgtcgtcccg gaggaggggg gccacctcgt tgagcatgtc
cctgacgcgc 5340atgttctccc cgaccagatc cgccagaagg cgctcgccgc ccagggacag
cagctcttgc 5400aaggaagcaa agtttttcag cggcttgagg ccgtccgccg tgggcatgtt
tttcagggtc 5460tggctcagca gctccaggcg gtcccagagc tcggtgacgt gctctacggc
atctctatcc 5520agcatatctc ctcgtttcgc gggttggggc gactttcgct gtagggcacc
aagcggtggt 5580cgtccagcgg ggccaaagtc atgtccttcc atgggcgcag ggtcctcgtc
agggtggtct 5640gggtcacggt gaaggggtgc gctccgggct gagcgcttgc caaggtgcgc
ttgaggctgg 5700ttctgctggt gctgaagcgc tgccggtctt cgccctgcgc gtcggccagg
tagcatttga 5760ccatggtgtc atagtccagc ccctccgcgg cgtgtccctt ggcgcgcagc
ttgcccttgg 5820aggtggcgcc gcacgagggg cagagcaggc tcttgagcgc gtagagcttg
ggggcgagga 5880agaccgattc gggggagtag gcgtccgcgc cgcagacccc gcacacggtc
tcgcactcca 5940ccagccaggt gagctcgggg cgcgccgggt caaaaaccag gtttccccca
tgctttttga 6000tgcgtttctt acctcgggtc tccatgaggt ggtgtccccg ctcggtgacg
aagaggctgt 6060ccgtgtctcc gtagaccgac ttgaggggtc ttttctccag gggggtccct
cggtcttcct 6120cgtagaggaa ctcggaccac tctgagacga aggcccgcgt ccaggccagg
acgaaggagg 6180ctatgtggga ggggtagcgg tcgttgtcca ctagggggtc caccttctcc
aaggtgtgaa 6240gacacatgtc gccttcctcg gcgtccagga aggtgattgg cttgtaggtg
taggccacgt 6300gaccgggggt tcctgacggg ggggtataaa agggggtggg ggcgcgctcg
tcgtcactct 6360cttccgcatc gctgtctgcg agggccagct gctggggtga gtattccctc
tcgaaggcgg 6420gcatgacctc cgcgctgagg ttgtcagttt ccaaaaacga ggaggatttg
atgttcacct 6480gtcccgaggt gatacctttg agggtacccg cgtccatctg gtcagaaaac
acgatctttt 6540tattgtccag cttggtggcg aacgacccgt agagggcgtt ggagagcagc
ttggcgatgg 6600agcgcagggt ctggttcttg tccctgtcgg cgcgctcctt ggccgcgatg
ttgagctgca 6660cgtactcgcg cgcgacgcag cgccactcgg ggaagacggt ggtgcgctcg
tcgggcacca 6720ggcgcacgcg ccagccgcgg ttgtgcaggg tgaccaggtc cacgctggtg
gcgacctcgc 6780cgcgcaggcg ctcgttggtc cagcagagac ggccgccctt gcgcgagcag
aaggggggca 6840gggggtcgag ctgggtctcg tccggggggt ccgcgtccac ggtgaaaacc
ccggggcgca 6900ggcgcgcgtc gaagtagtct atcttgcaac cttgcatgtc cagcgcctgc
tgccagtcgc 6960gggcggcgag cgcgcgctcg taggggttga gcggcgggcc ccagggcatg
gggtgggtga 7020gtgcggaggc gtacatgccg cagatgtcat agacgtagag gggctcccgc
aggaccccga 7080tgtaggtggg gtagcagcgg ccgccgcgga tgctggcgcg cacgtagtca
tacagctcgt 7140gcgagggggc gaggaggtcg gggcccaggt tggtgcgggc ggggcgctcc
gcgcggaaga 7200cgatctgcct gaagatggca tgcgagttgg aagagatggt ggggcgctgg
aagacgttga 7260agctggcgtc ctgcaggccg acggcgtcgc gcacgaagga ggcgtaggag
tcgcgcagct 7320tgtgtaccag ctcggcggtg acctgcacgt cgagcgcgca gtagtcgagg
gtctcgcgga 7380tgatgtcata tttagcctgc cccttctttt tccacagctc gcggttgagg
acaaactctt 7440cgcggtcttt ccagtactct tggatcggga aaccgtccgg ttccgaacgg
taagagccta 7500gcatgtagaa ctggttgacg gcctggtagg cgcagcagcc cttctccacg
gggagggcgt 7560aggcctgcgc ggccttgcgg agcgaggtgt gggtcagggc gaaggtgtcc
ctgaccatga 7620ctttgaggta ctggtgcttg aagtcggagt cgtcgcagcc gccccgctcc
cagagcgaga 7680agtcggtgcg cttcttggag cgggggttgg gcagagcgaa ggtgacatcg
ttgaagagga 7740ttttgcccgc gcggggcatg aagttgcggg tgatgcggaa gggccccggc
acttcagagc 7800ggttgttgat gacctgggcg gcgagcacga tctcgtcgaa gccgttgatg
ttgtggccca 7860cgatgtagag ttccaggaag cggggccggc cctttacggt gggcagcttc
tttagctctt 7920cgtaggtgag ctcctcgggc gaggcgaggc cgtgctcggc cagggcccag
tccgcgaggt 7980gcgggttgtc tctgaggaag gactcccaga ggtcgcgggc caggagggtc
tgcaggcggt 8040ccctgaaggt cctgaactgg cggcccacgg ccattttttc gggggtgatg
cagtagaagg 8100tgagggggtc ttgctgccag cggtcccagt cgagctgcag ggcgaggtcg
cgcgcggcgg 8160tgaccaggcg ctcgtcgccc ccgaatttca tgaccagcat gaagggcacg
agctgctttc 8220cgaaggcccc catccaagtg taggtctcta catcgtaggt gacaaagagg
cgctccgtgc 8280gaggatgcga gccgatcggg aagaactgga tctcccgcca ccagttggag
gagtggctgt 8340tgatgtggtg gaagtagaag tcccgtcgcc gggccgaaca ctcgtgctgg
cttttgtaaa 8400agcgagcgca gtactggcag cgctgcacgg gctgtacctc ctgcacgaga
tgcacctttc 8460gcccgcgcac gaggaagccg aggggaaatc tgagcccccc gcctggctcg
cggcatggct 8520ggtgctcttc tactttggat gcgtgtccgt ctccgtctgg ctcctcgagg
ggtgttacgg 8580tggagcggac caccacgccg cgcgagccgc aggtccagat atcggcgcgc
ggcggtcgga 8640gtttgatgac gacatcgcgc agctgggagc tgtccatggt ctggagctcc
cgcggcggcg 8700gcaggtcagc cgggagttct tgcaggttca cctcgcagag tcgggccagg
gcgcggggca 8760ggtctaggtg gtacctgatc tctaggggcg tgttggtggc ggcgtcgatg
gcttgcagga 8820gcccgcatcc ccggggggcg acgacggtgc cccgcggggt ggtggtggtg
gtggtggtgg 8880tggtggtggc ggtgcagctc agaagcggtg ccgcgggcgg gcccccggag
gtaggggggg 8940ctccggtccc gccggcaggg gcggcagcgg cacgtcggcg tggagcgcgg
gcaggagttg 9000gtgctgtgcc cggaggttgc tggcgaaggc gacgacgcgg cggttgatct
cctggatctg 9060gcgcctctgc gtgaagacga cgggcccggt gagcttgaac ctgaaagaga
gttcgacaga 9120atcaatctcg gtgtcattga ccgcggcctg gcgcaggatc tcctgcacgt
ctcccgagtt 9180gtcttggtag gcgatctcgg ccatgaactg ctcgatctct tcctcctgga
ggtctccgcg 9240tccggcgcgt tccacggtgg ccgccaggtc gttggagatg cgccccatga
gctgcgagaa 9300ggcgttgagt ccgccctcgt tccagactcg gctgtagacc acgcccccct
ggtcatcgcg 9360ggcgcgcatg accacctgcg cgaggttgag ctccacgtgc cgcgcgaaga
cggcgtagtt 9420gcgcagacgc tggaagaggt agttgagggt ggtggcggtg tgctcggcca
cgaagaagtt 9480catgacccag cggcgcaacg tggattcgtt gatgtccccc aaggcctcca
gccgttccat 9540ggcctcgtag aagtccacgg cgaagttgaa aaactgggag ttgcgcgccg
acacggtcaa 9600ctcctcctcc agaagacgga tgagctcggc gacggtgtcg cgcacctcgc
gctcgaaggc 9660tatggggatc tcttcctccg ctagcatcac cacctcctcc tcttcctcct
cttctggcac 9720ttccatgatg gcttcctcct cttcgggggg cggcggcggc ggcggtgggg
gagggggcgc 9780tctgcgccgg cggcggcgca ccgggaggcg gtccacgaag cgcgcgatca
tctccccgcg 9840gcggcggcgc atggtctcgg tgacggcgcg gccgttctcc cgggggcgca
gttggaagac 9900gccgccggac atctggtgct ggggcgggtg gccgtgaggc agcgaaacgg
cgctgacgat 9960gcatctcaac aattgctgcg taggtacgcc gccgagggac ctgagggagt
ccatatccac 10020cggatccgaa aacctttcga ggaaggcgtc taaccagtcg cagtcgcaag
gtaggctgag 10080caccgtggcg ggcggcgggg ggtgggggga gtgtctggcg gaggtgctgc
tgatgatgta 10140attgaagtag gcggacttga cacggcggat ggtcgacagg agcaccatgt
ccttgggtcc 10200ggcctgctgg atgcggaggc ggtcggctat gccccaggct tcgttctggc
atcggcgcag 10260gtccttgtag tagtcttgca tgagcctttc caccggcacc tcttctcctt
cctcttctgc 10320ttcttccatg tctgcttcgg ccctggggcg gcgccgcgcc cccctgcccc
ccatgcgcgt 10380gaccccgaac cccctgagcg gttggagcag ggccaggtcg gcgacgacgc
gctcggccag 10440gatggcctgc tgcacctgcg tgagggtggt ttggaagtca tccaagtcca
cgaagcggtg 10500gtaggcgccc gtgttgatgg tgtaggtgca gttggccatg acggaccagt
tgacggtctg 10560gtggcccggt tgcgacatct cggtgtacct gagtcgcgag taggcgcggg
agtcgaagac 10620gtagtcgttg caagtccgca ccaggtactg gtagcccacc aggaagtgcg
gcggcggctg 10680gcggtagagg ggccagcgca gggtggcggg ggctccgggg gccaggtctt
ccagcatgag 10740gcggtggtag gcgtagatgt acctggacat ccaggtgata cccgcggcgg
tggtggaggc 10800gcgcgggaag tcgcgcaccc ggttccagat gttgcgcagg ggcagaaagt
gctccatggt 10860aggcgtgctc tgtccagtca gacgcgcgca gtcgttgata ctctagacca
gggaaaacga 10920aagccggtca gcgggcactc ttccgtggtc tggtgaatag atcgcaaggg
tatcatggcg 10980gagggcctcg gttcgagccc cgggtccggg ccggacggtc cgccatgatc
cacgcggtta 11040ccgcccgcgt gtcgaaccca ggtgtgcgac gtcagacaac ggtggagtgt
tccttttggc 11100gtttttctgg ccgggcgccg gcgtcgcgta agagactaag ccgcgaaagc
gaaagcagta 11160agtggctcgc tccccgtagc cggagggatc cttgctaagg gttgcgttgc
ggcgaacccc 11220ggttcgaatc ccgtactcgg gccggccgga cccgcggcta aggtgttgga
ttggcctccc 11280cctcgtataa agaccccgct tgcggattga ctccggacac ggggacgagc
cccttttatt 11340tttgctttcc ccagatgcat ccggtgctgc ggcagatgcg ccccccgccc
cagcagcagc 11400aacaacacca gcaagagcgg cagcaacagc agcgggagtc atgcagggcc
ccctcaccca 11460ccctcggcgg gccggccacc tcggcgtccg cggccgtgtc tggcgcctgc
ggcggcggcg 11520gggggccggc tgacgacccc gaggagcccc cgcggcgcag ggccagacac
tacctggacc 11580tggaggaggg cgagggcctg gcgcggctgg gggcgccgtc tcccgagcgc
cacccgcggg 11640tgcagctgaa gcgcgactcg cgcgaggcgt acgtgcctcg gcagaacctg
ttcagggacc 11700gcgcgggcga ggagcccgag gagatgcggg acaggaggtt cagcgcaggg
cgggagctgc 11760ggcaggggct gaaccgcgag cggctgctgc gcgaggagga ctttgagccc
gacgcgcgga 11820cggggatcag ccccgcgcgc gcgcacgtgg cggccgccga cctggtgacg
gcgtacgagc 11880agacggtgaa ccaggagatc aacttccaaa agagtttcaa caaccacgtg
cgcacgctgg 11940tggcgcgcga ggaggtgacc atcgggctga tgcacctgtg ggactttgta
agcgcgctgg 12000tgcagaaccc caacagcaag cctctgacgg cgcagctgtt cctgatagtg
cagcacagca 12060gggacaacga ggcgtttagg gacgcgctgc tgaacatcac cgagcccgag
ggtcggtggc 12120tgctggacct gattaacatc ctgcagagca tagtggtgca ggagcgcagc
ctgagcctgg 12180ccgacaaggt ggcggccatc aactactcga tgctgagcct gggcaagttt
tacgcgcgca 12240agatctacca gacgccgtac gtgcccatag acaaggaggt gaagatcgac
ggtttttaca 12300tgcgcatggc gctgaaggtg ctcaccctga gcgacgacct gggcgtgtac
cgcaacgagc 12360gcatccacaa ggccgtgagc gtgagccggc ggcgcgagct gagcgaccgc
gagctgatgc 12420acagcctgca gcgggcgctg gcgggcgccg gcagcggcga cagggaggcg
gagtcctact 12480tcgatgcggg ggcggacctg cgctgggcgc ccagccggcg ggccctggag
gccgcggggg 12540tccgcgagga ctatgacgag gacggcgagg aggatgagga gtacgagcta
gaggagggcg 12600agtacctgga ctaaaccgcg ggtggtgttt ccggtagatg caagacccga
acgtggtgga 12660cccggcgctg cgggcggctc tgcagagcca gccgtccggc cttaactcct
cagacgactg 12720gcgacaggtc atggaccgca tcatgtcgct gacggcgcgt aacccggacg
cgttccggca 12780gcagccgcag gccaacaggc tctccgccat cctggaggcg gtggtgcctg
cgcgctcgaa 12840ccccacgcac gagaaggtgc tggccatagt gaacgcgctg gccgagaaca
gggccatccg 12900cccggacgag gccgggctgg tgtacgacgc gctgctgcag cgcgtggccc
gctacaacag 12960cggcaacgtg cagaccaacc tggaccggct ggtgggggac gtgcgcgagg
cggtggcgca 13020gcgcgagcgc gcggatcggc agggcaacct gggctccatg gtggcgctga
atgccttcct 13080gagcacgcag ccggccaacg tgccgcgggg gcaggaagac tacaccaact
ttgtgagcgc 13140gctgcggctg atggtgaccg agacccccca gagcgaggtg taccagtcgg
gcccggacta 13200cttcttccag accagcagac agggcctgca gacggtgaac ctgagccagg
ctttcaagaa 13260cctgcggggg ctgtggggcg tgaaggcgcc caccggcgac cgggcgacgg
tgtccagcct 13320gctgacgccc aactcgcgcc tgctgctgct gctgatcgcg ccgttcacgg
acagcggcag 13380cgtgtcccgg gacacctacc tggggcacct gctgaccctg taccgcgagg
ccatcgggca 13440ggcgcaggtg gacgagcaca ccttccagga gatcaccagc gtgagccgcg
cgctggggca 13500ggaggacacg agcagcctgg aggcgactct gaactacctg ctgaccaacc
ggcggcagaa 13560gattccctcg ctgcacagcc tgacctccga ggaggagcgc atcttgcgct
acgtgcagca 13620gagcgtgagc ctgaacctga tgcgcgacgg ggtgacgccc agcgtggcgc
tggacatgac 13680cgcgcgcaac atggaaccgg gcatgtacgc cgcgcaccgg ccttacatca
accgcctgat 13740ggactacctg catcgcgcgg cggccgtgaa ccccgagtac tttaccaacg
ccatcctgaa 13800cccgcactgg ctcccgccgc ccgggttcta cagcgggggc ttcgaggtcc
cggaggccaa 13860cgatggcttc ctgtgggacg acatggacga cagcgtgttc tccccgcggc
cgcaggcgct 13920ggcggaagcg tccctgctgc gtcccaagaa ggaggaggag gaggcgagtc
gccgccgcgg 13980cagcagcggc gtggcttctc tgtccgagct gggggcggca gccgccgcgc
gccccgggtc 14040cctgggcggc agcccctttc cgagcctggt ggggtctctg cacagcgagc
gcaccacccg 14100ccctcggctg ctgggcgagg acgagtacct gaataactcc ctgctgcagc
cggtgcggga 14160gaaaaacctg ccccccgcct tccccaacaa cgggatagag agcctggtgg
acaagatgag 14220cagatggaag acctatgcgc aggagcacag ggacgcgccc gcgctccggc
cgcccacgcg 14280gcgccagcgc cacgaccggc agcgggggct ggtgtgggat gacgaggact
ccgcggacga 14340tagcagcgtg ctggacctgg gagggagcgg caacccgttc gcgcacctgc
gcccccgcct 14400ggggaggatg ttttaaaaaa aaaaaaagca agaagcatga tgcaaaatta
aataaaactc 14460accaaggcca tggcgaccga gcgttggttt cttgtgttcc cttcagtatg
cggcgcgcgg 14520cgatgtacca ggagggacct cctccctctt acgagagcgt ggtgggcgcg
gcggcggcgg 14580cgccctcttc tccctttgcg tcgcagctgc tggagccgcc gtacgtgcct
ccgcgctacc 14640tgcggcctac gggggggaga aacagcatcc gttactcgga gctggcgccc
ctgttcgaca 14700ccacccgggt gtacctggtg gacaacaagt cggcggacgt ggcctccctg
aactaccaga 14760acgaccacag caattttttg accacggtca tccagaacaa tgactacagc
ccgagcgagg 14820ccagcaccca gaccatcaat ctggatgacc ggtcgcactg gggcggcgac
ctgaaaacca 14880tcctgcacac caacatgccc aacgtgaacg agttcatgtt caccaataag
ttcaaggcgc 14940gggtgatggt gtcgcgctcg cacaccaagg aagaccgggt ggagctgaag
tacgagtggg 15000tggagttcga gctgccagag ggcaactact ccgagaccat gaccattgac
ctgatgaaca 15060acgcgatcgt ggagcactat ctgaaagtgg gcaggcaaaa cggggtcctg
gagagcgaca 15120tcggggtcaa gttcgacacc aggaacttcc gcctggggct ggaccccgtg
accgggctgg 15180ttatgcccgg ggtgtacacc aacgaggcct tccatcccga catcatcctg
ctgcccggct 15240gcggggtgga cttcacttac agccgcctga gcaacctcct gggcatccgc
aagcggcagc 15300ccttccagga gggcttcagg atcacctacg aggacctgga ggggggcaac
atccccgcgc 15360tcctcgatgt ggaggcctac caggatagct tgaaggaaaa tgaggcggga
caggaggata 15420ccacccccgc cgcctccgcc gccgccgagc agggcgagga tgctgctgac
accgcggccg 15480cggacggggc agaggccgac cccgctatgg tggtggaggc tcccgagcag
gaggaggata 15540tgaatgacag tgcggtgcgc ggagacacct tcgtcacccg gggggaggaa
aagcaagcgg 15600aggccgaggc cgcggccgag gaaaagcaac tggcggcagc agcggcggcg
gcggcgttgg 15660ccgcggcgga ggctgagtct gaggggacca agcccgccaa ggagcccgtg
attaagcccc 15720tgaccgaaga tagcaagaag cgcagttaca acctgctcaa ggacagcacc
aacaccgcgt 15780accgcagctg gtacctggcc tacaactacg gcgacccgtc gacgggggtg
cgctcctgga 15840ccctgctgtg cacgccggac gtgacctgcg gctcggagca ggtgtactgg
tcgctgcccg 15900acatgatgca agaccccgtg accttccgct ccacgcggca ggtcagcaac
ttcccggtgg 15960tgggcgccga gctgctgccc gtgcactcca agagcttcta caacgaccag
gccgtctact 16020cccagctcat ccgccagttc acctctctga cccacgtgtt caatcgcttt
cctgagaacc 16080agattctggc gcgcccgccc gcccccacca tcaccaccgt cagtgaaaac
gttcctgctc 16140tcacagatca cgggacgcta ccgctgcgca acagcatcgg aggagtccag
cgagtgaccg 16200ttactgacgc cagacgccgc acctgcccct acgtttacaa ggccttgggc
atagtctcgc 16260cgcgcgtcct ttccagccgc actttttgag caacaccacc atcatgtcca
tcctgatctc 16320acccagcaat aactccggct ggggactgct gcgcgcgccc agcaagatgt
tcggaggggc 16380gaggaagcgt tccgagcagc accccgtgcg cgtgcgcggg cacttccgcg
ccccctgggg 16440agcgcacaaa cgcggccgcg cggggcgcac caccgtggac gacgccatcg
actcggtggt 16500ggagcaggcg cgcaactaca ggcccgcggt ctctaccgtg gacgcggcca
tccagaccgt 16560ggtgcggggc gcgcggcggt acgccaagct gaagagccgc cggaagcgcg
tggcccgccg 16620ccaccgccgc cgacccgggg ccgccgccaa acgcgccgcc gcggccctgc
ttcgccgggc 16680caagcgcacg ggccgccgcg ccgccatgag ggccgcgcgc cgcttggccg
ccggcatcac 16740cgccgccacc atggcccccc gtacccgaag acgcgcggcc gccgccgccg
ccgccgccat 16800cagtgacatg gccagcaggc gccggggcaa cgtgtactgg gtgcgcgact
cggtgaccgg 16860cacgcgcgtg cccgtgcgct tccgcccccc gcggacttga gatgatgtga
aaaaacaaca 16920ctgagtctcc tgctgttgtg tgtatcccag cggcggcggc gcgcgcagcg
tcatgtccaa 16980gcgcaaaatc aaagaagaga tgctccaggt cgtcgcgccg gagatctatg
ggcccccgaa 17040gaaggaagag caggattcga agccccgcaa gataaagcgg gtcaaaaaga
aaaagaaaga 17100tgatgacgat gccgatgggg aggtggagtt cctgcgcgcc acggcgccca
ggcgcccggt 17160gcagtggaag ggccggcgcg taaagcgcgt cctgcgcccc ggcaccgcgg
tggtcttcac 17220gcccggcgag cgctccaccc ggactttcaa gcgcgtctat gacgaggtgt
acggcgacga 17280agacctgctg gagcaggcca acgagcgctt cggagagttt gcttacggga
agcgtcagcg 17340ggcgctgggg aaggaggacc tgctggcgct gccgctggac cagggcaacc
ccacccccag 17400tctgaagccc gtgaccctgc agcaggtgct gccgagcagc gcaccctccg
aggcgaagcg 17460gggtctgaag cgcgagggcg gcgacctggc gcccaccgtg cagctcatgg
tgcccaagcg 17520gcagaggctg gaggatgtgc tggagaaaat gaaagtagac cccggtctgc
agccggacat 17580cagggtccgt cccatcaagc aggtggcgcc gggcctcggc gtgcagaccg
tggacgtggt 17640catccccacc ggcaactccc ccgccgccac caccactacc gctgcctcca
cggacatgga 17700gacacagacc gatcccgccg cagccgcagc cgccgccgca gccgcgacct
cctcggcgga 17760ggtgcagacg gacccctggc tgccgccggc gatgtcagct ccccgcgcgc
gccgcggacg 17820cagaaagtac ggcgccgcca acgcgctcct gcccgagtac gccttgcatc
cttccatcgc 17880gcccaccccc ggctaccgag gctataccta ccgcccgcga agagccaagg
gttccacccg 17940ccgtccccgc cgacgcgccg ccgccaccac ccgccgccgc cgccgcagac
gccagcccgc 18000actggctcca gtctccgtga ggagagtggc gcgcgacgga cacaccctgg
tgctgcccag 18060ggcgcgctac caccccagca tcgtttaaaa gcctgttgtg gttcttgcag
atatggccct 18120cacttgccgc ctccgtttcc cggtgccggg ataccgagga ggaagatcgc
gccgcaggag 18180gggtctggcc ggccgcggcc tgagcggagg cagccgccgc gcgcaccggc
ggcgacgcgc 18240caccagccga cgcatgcgcg gcggggtgct gcccctgtta atccccctga
tcgccgcggc 18300gatcggcgcc gtgcccggga tcgcctccgt ggccttgcaa gcgtcccaga
ggcattgaca 18360gacttgcaaa cttgcaaata tggaaaaaaa aaaaaaaccc caataaaaag
tctagactct 18420cacgctcgct tggtcctgtg actattttgt agaatggaag acatcaactt
tgcgtcgctg 18480gccccgcgtc acggctcgcg cccgttcctg ggacactgga acgatatcgg
caccagcaac 18540atgagcggtg gcgccttcag ttggggctct ctgtggagcg gcattaaaag
tatcgggtct 18600gccgttaaaa attacggctc ccgggcctgg aacagcagca cgggccagat
gttgagagac 18660aagttgaaag agcagaactt ccagcagaag gtggtggagg gcctggcctc
cggcatcaac 18720ggggtggtgg acctggccaa ccaggccgtg cagaataaaa tcaacagcag
actggacccc 18780cggccgccgg tggaggaggt gccgccggcg ctggagacgg tgtcccccga
tgggcgtggc 18840gagaagcgcc cgcggcccga tagggaagag accactctgg tcacgcagac
cgatgagccg 18900cccccgtatg aggaggccct aaagcaaggt ctgcccacca cgcggcccat
cgcgcccatg 18960gccaccgggg tggtgggccg ccacaccccc gccacgctgg acttgcctcc
gcccgccgat 19020gtgccgcagc agcagaaggc ggcacagccg ggcccgcccg cgaccgcctc
ccgttcctcc 19080gccggtcctc tgcgccgcgc ggccagcggc ccccgcgggg gggtcgcgag
gcacggcaac 19140tggcagagca cgctgaacag catcgtgggt ctgggggtgc ggtccgtgaa
gcgccgccga 19200tgctactgaa tagcttagct aacgtgttgt atgtgtgtat gcgccctatg
tcgccgccag 19260aggagctgct gagtcgccgc cgttcgcgcg cccaccacca ccgccactcc
gcccctcaag 19320atggcgaccc catcgatgat gccgcagtgg tcgtacatgc acatctcggg
ccaggacgcc 19380tcggagtacc tgagccccgg gctggtgcag ttcgcccgcg ccaccgagag
ctacttcagc 19440ctgagtaaca agtttaggaa ccccacggtg gcgcccacgc acgatgtgac
caccgaccgg 19500tctcagcgcc tgacgctgcg gttcattccc gtggaccgcg aggacaccgc
gtactcgtac 19560aaggcgcggt tcaccctggc cgtgggcgac aaccgcgtgc tggacatggc
ctccacctac 19620tttgacatcc gcggggtgct ggaccggggt cccactttca agccctactc
tggcaccgcc 19680tacaactccc tggcccccaa gggcgctccc aactcctgcg agtgggagca
agaggaaact 19740caggcagttg aagaagcagc agaagaggaa gaagaagatg ctgacggtca
agctgaggaa 19800gagcaagcag ctaccaaaaa gactcatgta tatgctcagg ctcccctttc
tggcgaaaaa 19860attagtaaag atggtctgca aataggaacg gacgctacag ctacagaaca
aaaacctatt 19920tatgcagacc ctacattcca gcccgaaccc caaatcgggg agtcccagtg
gaatgaggca 19980gatgctacag tcgccggcgg tagagtgcta aagaaatcta ctcccatgaa
accatgctat 20040ggttcctatg caagacccac aaatgctaat ggaggtcagg gtgtactaac
ggcaaatgcc 20100cagggacagc tagaatctca ggttgaaatg caattctttt caacttctga
aaacgcccgt 20160aacgaggcta acaacattca gcccaaattg gtgctgtata gtgaggatgt
gcacatggag 20220accccggata cgcacctttc ttacaagccc gcaaaaagcg atgacaattc
aaaaatcatg 20280ctgggtcagc agtccatgcc caacagacct aattacatcg gcttcagaga
caactttatc 20340ggcctcatgt attacaatag cactggcaac atgggagtgc ttgcaggtca
ggcctctcag 20400ttgaatgcag tggtggactt gcaagacaga aacacagaac tgtcctacca
gctcttgctt 20460gattccatgg gtgacagaac cagatacttt tccatgtgga atcaggcagt
ggacagttat 20520gacccagatg ttagaattat tgaaaatcat ggaactgaag acgagctccc
caactattgt 20580ttccctctgg gtggcatagg ggtaactgac acttaccagg ctgttaaaac
caacaatggc 20640aataacgggg gccaggtgac ttggacaaaa gatgaaactt ttgcagatcg
caatgaaata 20700ggggtgggaa acaatttcgc tatggagatc aacctcagtg ccaacctgtg
gagaaacttc 20760ctgtactcca acgtggcgct gtacctacca gacaagctta agtacaaccc
ctccaatgtg 20820gacatctctg acaaccccaa cacctacgat tacatgaaca agcgagtggt
ggccccgggg 20880ctggtggact gctacatcaa cctgggcgcg cgctggtcgc tggactacat
ggacaacgtc 20940aaccccttca accaccaccg caatgcgggc ctgcgctacc gctccatgct
cctgggcaac 21000gggcgctacg tgcccttcca catccaggtg ccccagaagt tctttgccat
caagaacctc 21060ctcctcctgc cgggctccta cacctacgag tggaacttca ggaaggatgt
caacatggtc 21120ctccagagct ctctgggtaa cgatctcagg gtggacgggg ccagcatcaa
gttcgagagc 21180atctgcctct acgccacctt cttccccatg gcccacaaca cggcctccac
gctcgaggcc 21240atgctcagga acgacaccaa cgaccagtcc ttcaatgact acctttccgc
cgccaacatg 21300ctctacccca tacccgccaa cgccaccaac gtccccatct ccatcccctc
gcgcaactgg 21360gcggccttcc gcggctgggc cttcacccgc ctcaagacca aggagacccc
ctccctgggc 21420tcgggattcg acccctacta cacctactcg ggctctattc cctacctgga
cggcaccttc 21480tacctcaacc acactttcaa gaaggtctcg gtcaccttcg actcctcggt
cagctggccg 21540ggcaacgacc gtctgctcac ccccaacgag ttcgagatca agcgctcggt
cgacggggaa 21600ggctacaacg tggcccagtg caacatgacc aaggactggt tcctggtcca
gatgctggcc 21660aactacaaca tcggctacca gggcttctac atcccagaga gctacaagga
caggatgtac 21720tccttcttca ggaacttcca gcccatgagc cggcaggtgg tggaccagac
caagtacaag 21780gactaccagg aggtgggcat catccaccag cacaacaact cgggcttcgt
gggctacctc 21840gcccccacca tgcgcgaggg acaggcctac cccgccaact tcccctaccc
gctcataggc 21900aagaccgcgg tcgacagcat cacccagaaa aagttcctct gcgaccgcac
cctctggcgc 21960atccccttct ccagcaactt catgtccatg ggtgcgctct cggacctggg
ccagaacttg 22020ctctacgcca actccgccca cgccctcgac atgaccttcg aggtcgaccc
catggacgag 22080cccacccttc tctatgttct gttcgaagtc tttgacgtgg tccgggtcca
ccagccgcac 22140cgcggcgtca tcgagaccgt gtacctgcgt acgcccttct cggccggcaa
cgccaccacc 22200taaagaagca agccgcagtc atcgccgcct gcatgccgtc gggttccacc
gagcaagagc 22260tcagggccat cgtcagagac ctgggatgcg ggccctattt tttgggcacc
ttcgacaagc 22320gcttccctgg ctttgtctcc ccacacaagc tggcctgcgc catcgtcaac
acggccggcc 22380gcgagaccgg gggcgtgcac tggctggcct ttgcctggaa cccgcgctcc
aaaacatgct 22440tcctctttga ccccttcggc ttttcggacc agcggctcaa gcaaatctac
gagttcgagt 22500acgagggctt gctgcgtcgc agcgccatcg cctcctcgcc cgaccgctgc
gtcaccctcg 22560aaaagtccac ccagaccgtg caggggcccg actcggccgc ctgcggtctc
ttctgctgca 22620tgtttctgca cgcctttgtg cactggcctc agagtcccat ggaccgcaac
cccaccatga 22680acttgctgac gggggtgccc aactccatgc tccaaagccc ccaggtcgag
cccaccctgc 22740gccgcaacca ggagcagctc tacagcttcc tggagcgcca ctcgccctac
ttccgccgcc 22800acagcgcaca gatcaggagg gccacctcct tctgccactt gcaagagatg
caagaagggt 22860aataacgatg tacacacttt tttctcaata aatggcattt tttttttatt
tatacaagct 22920ctctggggta ttcatttccc accaccacca cccgccgttg tcgccatctg
gctctattta 22980gaaatcgaaa gggttctgcc gggagtcgcc gtgcgccacg ggcagggaca
cgttgcgata 23040ctggtagcgg gtgccccact tgaactcggg caccaccagg cgaggcagct
cggggaagtt 23100ttcgctccac aggctgcggg tcagcaccag cgcgttcatc aggtcgggcg
ccgagatctt 23160gaagtcgcag ttggggccgc cgccctgcgc gcgcgagttg cggtacaccg
ggttgcagca 23220ctggaacacc aacagcgccg ggtgcttcac gctggccagc acgctgcggt
cggagatcag 23280ctcggcgtcc aggtcctccg cgttgctcag cgcgaacggg gtcatcttgg
gcacttgccg 23340ccccaggaag ggcgcgtgcc ccggtttcga gttgcagtcg cagcgcagcg
ggatcagcag 23400gtgcccgtgc ccggactcgg cgttggggta cagcgcgcgc atgaaggcct
gcatctggcg 23460gaaggccatc tgggccttgg cgccctccga gaagaacatg ccgcaggact
tgcccgagaa 23520ctggtttgcg gggcagctgg cgtcgtgcag gcagcagcgc gcgtcggtgt
tggcgatctg 23580caccacgttg cgcccccacc ggttcttcac gatcttggcc ttggacgatt
gctccttcag 23640cgcgcgctgc ccgttctcgc tggtcacatc catctcgatc acatgttcct
tgttcaccat 23700gctgctgccg tgcagacact tcagctcgcc ctccgtctcg gtgcagcggt
gctgccacag 23760cgcgcagccc gtgggctcga aagacttgta ggtcacctcc gcgaaggact
gcaggtaccc 23820ctgcaaaaag cggcccatca tggtcacgaa ggtcttgttg ctgctgaagg
tcagctgcag 23880cccgcggtgc tcctcgttca gccaggtctt gcacacggcc gccagcgcct
ccacctggtc 23940gggcagcatc ttgaagttca ccttcagctc attctccacg tggtacttgt
ccatcagcgt 24000gcgcgccgcc tccatgccct tctcccaggc cgacaccagc ggcaggctca
cggggttctt 24060caccatcacc gtggccgccg cctccgccgc gctttcgctt tccgccccgc
tgttctcttc 24120ctcttcctcc tcttcctcgc cgccgcccac tcgcagcccc cgcaccacgg
ggtcgtcttc 24180ctgcaggcgc tgcaccttgc gcttgccgtt gcgcccctgc ttgatgcgca
cgggcgggtt 24240gctgaagccc accatcacca gcgcggcctc ttcttgctcg tcctcgctgt
ccagaatgac 24300ctccggggag ggggggttgg tcatcctcag taccgaggca cgcttctttt
tcttcctggg 24360ggcgttcgcc agctccgcgg ctgcggccgc tgccgaggtc gaaggccgag
ggctgggcgt 24420gcgcggcacc agcgcgtctt gcgagccgtc ctcgtcctcc tcggactcga
gacggaggcg 24480ggcccgcttc ttcgggggcg cgcggggcgg cggaggcggc ggcggcgacg
gagacgggga 24540cgagacatcg tccagggtgg gtggacggcg ggccgcgccg cgtccgcgct
cgggggtggt 24600ttcgcgctgg tcctcttccc gactggccat ctcccactgc tccttctcct
ataggcagaa 24660agagatcatg gagtctctca tgcgagtcga gaaggaggag gacagcctaa
ccgccccctc 24720tgagccctcc accaccgccg ccaccaccgc caatgccgcc gcggacgacg
cgcccaccga 24780gaccaccgcc agtaccaccc tccccagcga cgcacccccg ctcgagaatg
aagtgctgat 24840cgagcaggac ccgggttttg tgagcggaga ggaggatgag gtggatgaga
aggagaagga 24900ggaggtcgcc gcctcagtgc caaaagagga taaaaagcaa gaccaggacg
acgcagataa 24960ggatgagaca gcagtcgggc gggggaacgg aagccatgat gctgatgacg
gctacctaga 25020cgtgggagac gacgtgctgc ttaagcacct gcaccgccag tgcgtcatcg
tctgcgacgc 25080gctgcaggag cgctgcgaag tgcccctgga cgtggcggag gtcagccgcg
cctacgagcg 25140gcacctcttc gcgccgcacg tgccccccaa gcgccgggag aacggcacct
gcgagcccaa 25200cccgcgtctc aacttctacc cggtcttcgc ggtacccgag gtgctggcca
cctaccacat 25260cttcttccaa aactgcaaga tccccctctc ctgccgcgct aaccgcaccc
gcgccgacaa 25320aaccctgacc ctgcggcagg gcgcccacat acctgatatt gcctctctgg
aggaagtgcc 25380caagatcttc gagggtctcg gtcgcgacga gaaacgggcg gcgaacgctc
tgcacggaga 25440cagcgaaaac gagagtcact cgggggtgct ggtggagctc gagggcgaca
acgcgcgcct 25500ggccgtactc aagcgcagca tagaggtcac ccactttgcc tacccggcgc
tcaacctgcc 25560ccccaaggtc atgagtgtgg tcatgggcga gctcatcatg cgccgcgctc
agcccctggc 25620cgcggatgca aacttgcaag agtcctccga ggaaggcctg cccgcggtca
gcgacgagca 25680gctagcgcgc tggctggaga cccgcgaccc cgcgcagctg gaggagcggc
gcaagctcat 25740gatggccgcg gtgctggtca ccgtggagct cgagtgtctg cagcgcttct
tcgcggaccc 25800cgagatgcag cgcaagctcg aggagaccct gcactacacc ttccgccagg
gctacgtgcg 25860ccaggcctgc aagatctcca acgtggagct ctgcaacctg gtctcctacc
tgggcatcct 25920gcacgagaac cgcctcgggc agaacgtcct gcactccacc ctcaaagggg
aggcgcgccg 25980cgactacatc cgcgactgcg cctacctctt cctctgctac acctggcaga
cggccatggg 26040ggtctggcag cagtgcctgg aggagcgcaa cctcaaggag ctggaaaagc
tactcaagcg 26100caccctcagg gacctctgga cgggcttcaa cgagcgctcg gtggccgccg
cgctggcgga 26160catcatcttc cccgagcgcc tgctcaagac cctgcagcag ggcctgcccg
acttcaccag 26220ccagagcatg ctgcagaact ttaggacttt catcctggag cgctcgggca
tcctgcctgc 26280cacttgctgc gcgctgccca gcgacttcgt gcccatcaag tacagggagt
gcccgccgcc 26340gctctggggc cactgctacc tcttccagct ggccaactac ctcgcctacc
actcggacct 26400catggaagac gtgagcggcg agggcctgct cgagtgccac tgccgctgca
acctctgcac 26460gccccaccgc tctctagtct gcaacccgca gctgctcagc gagagtcaga
ttatcggtac 26520cttcgagctg cagggtccct cgcctgacga gaagtccgcg gctccggggc
tgaaactcac 26580tccggggctg tggacttccg cctacctacg caaatttgta cctgaggact
accacgccca 26640cgagatcagg ttctacgaag accaatcccg cccgcccaag gcggagctca
ccgcctgcgt 26700catcacccag gggcacatcc tgggccaatt gcaagccatc aacaaagccc
gccgagagtt 26760cttgctgaaa aagggtcggg gggtgtacct ggacccccag tccggcgagg
agctaaaccc 26820gctacccccg ccgccgcccc agcagcggga ccttgcttcc caggatggca
cccagaaaga 26880agcagcagcc gccgccgccg cagccataca tgcttctgga ggaagaggag
gaggactggg 26940acagtcaggc agaggaggtt tcggacgagg agcaggagga gatgatggaa
gactgggagg 27000aggacagcag cctagacgag gaagcttcag aggccgaaga ggtggcagac
gcaacaccat 27060caccctcggt cgcagccccc tcgccggggc ccctgaaatc ctccgaaccc
agcaccagcg 27120ctataacctc cgctcctccg gcgccggcgc cacccgcccg cagacccaac
cgtagatggg 27180acaccacagg aaccggggtc ggtaagtcca agtgcccgcc gccgccaccg
cagcagcagc 27240agcagcgcca gggctaccgc tcgtggcgcg ggcacaagaa cgccatagtc
gcctgcttgc 27300aagactgcgg gggcaacatc tctttcgccc ggcgcttcct gctattccac
cacggggtcg 27360cctttccccg caatgtcctg cattactacc gtcatctcta cagcccctac
tgcagcggcg 27420acccagaggc ggcagcggca gccacagcgg cgaccaccac ctaggaagat
atcctccgcg 27480ggcaagacag cggcagcagc ggccaggaga cccgcggcag cagcggcggg
agcggtgggc 27540gcactgcgcc tctcgcccaa cgaacccctc tcgacccggg agctcagaca
caggatcttc 27600cccactttgt atgccatctt ccaacagagc agaggccagg agcaggagct
gaaaataaaa 27660aacagatctc tgcgctccct cacccgcagc tgtctgtatc acaaaagcga
agatcagctt 27720cggcgcacgc tggaggacgc ggaggcactc ttcagcaaat actgcgcgct
cactcttaaa 27780gactagctcc gcgcccttct cgaatttagg cgggagaaaa ctacgtcatc
gccggccgcc 27840gcccagcccg cccagccgag atgagcaaag agattcccac gccatacatg
tggagctacc 27900agccgcagat gggactcgcg gcgggagcgg cccaggacta ctccacccgc
atgaactaca 27960tgagcgcggg accccacatg atctcacagg tcaacgggat ccgcgcccag
cgaaaccaaa 28020tactgctgga acaggcggcc atcaccgcca cgccccgcca taatctcaac
ccccgaaatt 28080ggcccgccgc cctcgtgtac caggaaaccc cctccgccac caccgtacta
cttccgcgtg 28140acgcccaggc cgaagtccag atgactaact caggggcgca gctcgcgggc
ggctttcgtc 28200acggggcgcg gccgctccga ccaggtataa gacacctgat gatcagaggc
cgaggtatcc 28260agctcaacga cgagtcggtg agctcttcgc tcggtctccg tccggacgga
actttccagc 28320tcgccggatc cggccgctct tcgttcacgc cccgccaggc gtacctgact
ctgcagacct 28380cgtcctcgga gccccgctcc ggaggcatcg gaaccctcca gttcgtggag
gagttcgtgc 28440cctcggtcta cttcaacccc ttctcgggac ctcccggacg ctaccccgac
cagttcattc 28500cgaactttga cgcggtgaag gactcggcgg acggctacga ctgaatgtca
ggtgccgagg 28560cagagcagct tcgcctgaga cacctcgagc actgccgccg ccacaagtgc
ttcgcccgcg 28620gttccggtga gttctgctac tttcagctac ccgaggagca taccgagggg
ccggcgcacg 28680gcgtccgcct gaccacccag ggcgaggtta cctgttccct catccgggag
ttcaccctcc 28740gtcccctgct agtggagcgg gagcggggtc cctgtgtcct aactatcgcc
tgcaactgcc 28800ctaaccctgg attacatcaa gatctttgct gtcatctctg tgctgagttt
aataaacgct 28860gagatcagaa tctactgggg ctcctgtcgc catcctgtga acgccaccgt
cttcacccac 28920cccgaccagg cccaggcgaa cctcacctgc ggtctgcatc ggagggccaa
gaagtacctc 28980acctggtact tcaacggcac cccctttgtg gtttacaaca gcttcgacgg
ggacggagtc 29040tccctgaaag accagctctc cggtctcagc tactccatcc acaagaacac
caccctccaa 29100ctcttccctc cctacctgcc gggaacctac gagtgcgtca ccggccgctg
cacccacctc 29160acccgcctga tcgtaaacca gagctttccg ggaacagata actccctctt
ccccagaaca 29220ggaggtgagc tcaggaaact ccccggggac cagggcggag acgtaccttc
gacccttgtg 29280gggttaggat tttttattac cgggttgctg gctcttttaa tcaaagcttc
cttgagattt 29340gttctttcct tctacgtgta tgaacacctc agcctccaat aactctaccc
tttcttcgga 29400atcaggtgac ttctctgaaa tcgggcttgg tgtgctgctt actctgttga
tttttttcct 29460tatcatactc agccttctgt gcctcaggct cgccgcctgc tgcgcacaca
tctatatcta 29520ctgctggttg ctcaagtgca ggggtcgcca cccaagatga acaggtacat
ggtcctatcg 29580atcctaggcc tgctggccct ggcggcctgc agcgccgcca aaaaagagat
tacctttgag 29640gagcccgctt gcaatgtaac tttcaagccc gagggtgacc aatgcaccac
cctcgtcaaa 29700tgcgttacca atcatgagag gctgcgcatc gactacaaaa acaaaactgg
ccagtttgcg 29760gtctatagtg tgtttacgcc cggagacccc tctaactact ctgtcaccgt
cttccagggc 29820ggacagtcta agatattcaa ttacactttc cctttttatg agttatgcga
tgcggtcatg 29880tacatgtcaa aacagtacaa cctgtggcct ccctctcccc aggcgtgtgt
ggaaaatact 29940gggtcttact gctgtatggc tttggcaatc actacgctcg ctctaatctg
cacggtgcta 30000tacataaaat tcaggcagag gcgaatcttt atcgatgaaa agaaaatgcc
ttgatcgcta 30060acaccggctt tctatctgca gaatgaatgc aatcacctcc ctactaatca
ccaccaccct 30120ccttgcgatt gcccatgggt tgacacgaat cgaagtgcca gtggggtcca
atgtcaccat 30180ggtgggcccc gccggcaatt ccaccctcat gtgggaaaaa tttgtccgca
atcaatgggt 30240tcatttctgc tctaaccgaa tcagtatcaa gcccagagcc atctgcgatg
ggcaaaatct 30300aactctgatc aatgtgcaaa tgatggatgc tgggtactat tacgggcagc
ggggagaaat 30360cattaattac tggcgacccc acaaggacta catgctgcat gtagtcgagg
cacttcccac 30420taccaccccc actaccacct ctcccaccac cactaccacc actactacta
ctactactac 30480cactaccgct gcccgccata cccgcaaaag caccatgatt agcacaaagc
cccctcgtgc 30540tcactcccac gccggcgggc ccatcggtgc gacctcagaa accaccgagc
tttgcttctg 30600ccaatgcact aacgccagcg ctcatgaact gttcgacctg gagaatgagg
atgcccagca 30660gagctccgct tgcctgaccc aggaggctgt ggagcccgtt gccctgaagc
agatcggtga 30720ttcaataatt gactcttctt cttttgccac tcccgaatac cctcccgatt
ctactttcca 30780catcacgggt accaaagacc ctaacctctc tttctacctg atgctgctgc
tctgtatctc 30840tgtggtctct tccgcgctga tgttactggg gatgttctgc tgcctgatct
gccgcagaaa 30900gagaaaagct cgctctcagg gccaaccact gatgcccttc ccctaccccc
cggattttgc 30960agataacaag atatgagctc gctgctgaca ctaaccgctt tactagcctg
cgctctaacc 31020cttgtcgctt gcgactcgag attccacaat gtcacagctg tggcaggaga
aaatgttact 31080ttcaactcca cggccgatac ccagtggtcg tggagtggct caggtagcta
cttaactatc 31140tgcaatagct ccacttcccc cagcatatcc ccaaccaagt accaatgcaa
tgccagcctg 31200ttcaccctca tcaacgcttc caccctggac aatggactct atgtaggcta
tgtacccttt 31260ggtgggcaag gaaagaccca cgcttacaac ctggaagttc gccagcccag
aaccactacc 31320caagcttctc ccaccaccac caccaccacc accaccacca tcaccagcag
cagcagcagc 31380cacagcagca gcagcagatt attgactttg gttttggcca gctcatctgc
cgctacccag 31440gccatctaca gctctgtgcc cgaaaccact cagatccacc gcccagaaac
gaccaccgcc 31500accaccctac acacctccag cgatcagatg ccgaccaaca tcaccccctt
ggctcttcaa 31560atgggactta caagccccac tccaaaacca gtggatgcgg ccgaggtctc
cgccctcgtc 31620aatgactggg cggggctggg aatgtggtgg ttcgccatag gcatgatggc
gctctgcctg 31680cttctgctct ggctcatctg ctgcctccac cgcaggcgag ccagaccccc
catctataga 31740cccatcattg tcctgaaccc cgataatgat gggatccata gattggatgg
cctgaaaaac 31800ctactttttt cttttacagt atgataaatt gagacatgcc tcgcattttc
ttgtacatgt 31860tccttctccc accttttctg gggtgttcta cgctggccgc tgtgtctcac
ctggaggtag 31920actgcctctc acccttcact gtctacctgc tttacggatt ggtcaccctc
actctcatct 31980gcagcctaat cacagtaatc atcgccttca tccagtgcat tgattacatc
tgtgtgcgcc 32040tcgcatactt cagacaccac ccgcagtacc gagacaggaa cattgcccaa
cttctaagac 32100tgctctaatc atgcataaga ctgtgatctg ccttctgatc ctctgcatcc
tgcccaccct 32160cacctcctgc cagtacacca caaaatctcc gcgcaaaaga catgcctcct
gccgcttcac 32220ccaactgtgg aatataccca aatgctacaa cgaaaagagc gagctctccg
aagcttggct 32280gtatggggtc atctgtgtct tagttttctg cagcactgtc tttgccctca
tgatctaccc 32340ctactttgat ttgggatgga acgcgatcga tgccatgaat taccccacct
ttcccgcacc 32400cgagataatt ccactgcgac aagttgtacc cgttgtcgtt aatcaacgcc
ccccatcccc 32460tacgcccact gaaatcagct actttaacct aacaggcgga gatgactgac
gccctagatc 32520tagaaatgga cggcatcagt accgagcagc gtctcctaga gaggcgcagg
caggcggctg 32580agcaagagcg cctcaatcag gagctccgag atctcgttaa cctgcaccag
tgcaaaagag 32640gcatcttttg tctggtaaag caggccaaag tcacctacga gaagaccggc
aacagccacc 32700gcctcagtta caaattgccc acccagcgcc agaagctggt gctcatggtg
ggtgagaatc 32760ccatcaccgt cacccagcac tcggtagaga ccgaggggtg tctgcactct
ccctgtcggg 32820gtccagaaga cctctgcacc ctggtaaaga ccctgtgcgg tctcagagat
ttagtcccct 32880ttaactaatc aaacactgga atcaataaaa agaatcactt acttaaaatc
agacagcagg 32940tctctgtcca gtttattcag cagcacctcc ttcccctcct cccaactctg
gtactccaaa 33000cgccttctgg cggcaaactt cctccacacc ctgaagggaa tgtcagattc
ttgctcctgt 33060ccctccgcac ccactatctt catgttgttg cagatgaagc gcaccaaaac
gtctgacgag 33120agcttcaacc ccgtgtaccc ctatgacacg gaaagcggcc ctccctccgt
ccctttcctc 33180acccctccct tcgtgtctcc cgatggattc caagaaagcc cccccggggt
cctgtctctg 33240aacctggccg agcccctggt cacttcccac ggcatgctcg ccctgaaaat
gggaagtggc 33300ctctccctgg acgacgctgg caacctcacc tctcaagata tcaccaccgc
tagccctccc 33360ctcaaaaaaa ccaagaccaa cctcagccta gaaacctcat cccccctaac
tgtaagcacc 33420tcaggcgccc tcaccgtagc agccgccgct cccctggcag tggccggcac
ctccctcacc 33480atgcaatcag aggcccccct gacagtacag gatgcaaaac tcaccctggc
caccaaaggc 33540cccctgaccg tgtctgaagg caaactggcc ttgcaaacat cggccccgct
gacggccgct 33600gacagcagca ccctcaccgt tagcgccaca ccaccaatta atgtaagcag
tggaagttta 33660ggcttagaca tggaagaccc tatgtatact cacgatggaa aactgggaat
aagaattggg 33720ggtccactaa gagtagtaga cagcttgcac acactcactg tagttaccgg
aaatggacta 33780actgtagata acaatgccct ccaaactaga gttacgggcg ccctaggtta
tgacacatca 33840ggaaatctac aattgagagc tgcaggaggt atgcgaattg atgcaaatgg
ccaacttatc 33900cttaatgtgg catacccatt tgatgctcag aacaatctca gccttagact
tggtcaggga 33960cccctgtata taaacacaga ccacaacctg gatttgaatt gcaacagagg
tctaaccaca 34020actaccacca acaacacaaa aaaacttgag actaaaatta gctcaggctt
agactatgac 34080accaatggtg ctgtcattat taaacttggc actggtctaa gcttcgacaa
cacaggcgcc 34140ctaactgtgg gaaacactgg tgatgataaa ctgactctgt ggacgacccc
agacccatct 34200ccaaattgca gaattcactc agacaaagac tgcaagttta ctctagtcct
aactaagtgt 34260ggaagccaaa tcctggcctc tgtcgccgcc ctagcggtat caggaaatct
ggcttcgata 34320acaggcaccg ttgccagcgt taccatcttt ctcagatttg atcagaatgg
agtgcttatg 34380gaaaactcct cgctagacag gcagtactgg aacttcagaa atggcaactc
aactaacgct 34440gccccctaca ccaatgcagt tgggttcatg ccaaacctcg cagcataccc
caaaacgcaa 34500agccagactg ctaaaaacaa cattgtaagt caggtttact tgaatggaga
caaatccaaa 34560cccatgaccc ttaccatcac cctcaatgga actaatgaat ccagtgaaac
tagccaggtg 34620agtcactact ccatgtcatt tacatgggct tgggaaagtg ggcaatatgc
cactgaaacc 34680tttgccacca actccttcac cttttcttac attgctgaac aataaaaagc
atgacactga 34740tgttcatttc tgattcttat tttattattt tcaaacacaa caaaatcatt
caagtcattc 34800ttccatctta gcttaataga cacagtagct taatagaccc agtagtgcaa
agccccattc 34860tagcttataa ctagtggaga agtactcgcc tacatggggg tagagtcata
atcgtgcatc 34920aggatagggc ggtggtgctg cagcagcgcg cgaataaact gctgccgccg
ccgctccgtc 34980ctgcaggaat acaacatggc agtggtctcc tcagcgatga ttcgcaccgc
ccgcagcata 35040aggcgccttg tcctccgggc acagcagcgc accctgatct cacttaaatc
agcacagtaa 35100ctgcagcaca gcaccacaat attgttcaaa atcccacagt gcaaggcgct
gtatccaaag 35160ctcatggcgg ggaccacaga acccacgtgg ccatcatacc acaagcgcag
gtagattaag 35220tggcgacccc tcataaacac gctggacata aacattacct cttttggcat
gttgtaattc 35280accacctccc ggtaccatat aaacctctga ttaaacatgg cgccatccac
caccatccta 35340aaccagctgg ccaaaacctg cccgccggct atacactgca gggaaccggg
actggaacaa 35400tgacagtgga gagcccagga ctcgtaacca tggatcatca tgctcgtcat
gatatcaatg 35460ttggcacaac acaggcacac gtgcatacac ttcctcagga ttacaagctc
ctcccgcgtt 35520agaaccatat cccagggaac aacccattcc tgaatcagcg taaatcccac
actgcaggga 35580agacctcgca cgtaactcac gttgtgcatt gtcaaagtgt tacattcggg
cagcagcgga 35640tgatcctcca gtatggtagc gcgggtttct gtctcaaaag gaggtagacg
atccctactg 35700tacggagtgc gccgagacaa ccgagatcgt gttggtcgta gtgtcatgcc
aaatggaacg 35760ccggacgtag tcatatttcc tgaagtctta gatctctcaa cgcagcacca
gcaccaacac 35820ttcgcagtgt aaaaggccaa gtgccgagag agtatatata ggaataaaaa
gtgacgtaaa 35880cgggcaaagt ccaaaaaacg cccagaaaaa ccgcacgcga acctacgccc
cgaaacgaaa 35940gccaaaaaac actagacact cccttccggc gtcaacttcc gctttcccac
gctacgtcac 36000ttgccccagt caaacaaact acatatcccg aacttccaag tcgccacgcc
caaaacaccg 36060cctacacctc cccgcccgcc ggcccgcccc caaacccgcc tcccgccccg
cgccccgccc 36120cgcgccgccc atctcattat catattggct tcaatccaaa ataaggtata
ttattgatga 36180tg
361821436643DNAPan troglodytes 14catcatcaat aatatacctc
aaacttttgg tgcgcgttaa tatgcaaatg aggtgtttga 60atttggggat gcggggcgct
gattggctga gagacgggcg accgttaggg gcggggcggg 120tgacgttttg atgacgtggc
cgtgaggcgg agccggtttg caagttctcg tgggaaaagt 180gacgtcaaac gaggtgtggt
ttgaacacgg aaatactcaa ttttcccgcg ctctctgaca 240ggaaatgagg tgtttctggg
cggatgcaag tgaaaacggg ccattttcgc gcgaaaactg 300aatgaggaag tgaaaatctg
agtaattccg cgtttatggc agggaggagt atttgccgag 360ggccgagtag actttgaccg
attacgtggg ggtttcgatt accgtatttt tcacctaaat 420ttccgcgtac ggtgtcaaag
tccggtgttt ttacgtaggc gtcagctgat cgccagggta 480tttaaacctg cgctctctag
tcaagaggcc actcttgagt gccagcgagt agagttttct 540cctccgcgcc gcgagtcaga
tctacacttt gaaagatgag gcacctgaga gacctgcccg 600gtaatgtttt cctggctact
gggaacgaga ttctggaact ggtggtggac gccatgatgg 660gtgacgaccc tcccgagccc
cctaccccat ttgaggcgcc ttcgctgtac gatttgtatg 720atctggaggt ggatgtgccc
gagaacgacc ccaacgagga ggcggtgaat gatttgttta 780gcgatgccgc gctgctggct
gccgagcagg ctaatacgga ctctggctca gacagcgatt 840cctctcttca taccccgaga
cccggcagag gtgagaaaaa gatccccgag cttaaagggg 900aagagctcga cctgcgctgc
tatgaggaat gcttgcctcc gagcgatgat gaggaggacg 960aggaggcgat tcgagctgca
gcgagcgagg gagtgaaagt tgcgggcgag agctttagcc 1020tggactgtcc tactctgccc
ggacacggct gtaagtcttg tgaatttcat cgcatgaata 1080ctggagataa gaatgtgatg
tgtgccctgt gctatatgag agcttacaac cattgtgttt 1140acagtaagtg tgattaactt
tagttgggaa ggcagagggt gactgggtgc tgactggttt 1200atttatgtat atgttttttt
atgtgtaggt cccgtctctg acgtagatga gacccccact 1260tcagagtgca tttcatcacc
cccagaaatt ggcgaggaac cgcccgaaga tattattcat 1320agaccagttg cagtgagagt
caccgggcgg agagcagctg tggagagttt ggatgacttg 1380ctacagggtg gggatgaacc
tttggacttg tgtacccgga aacgccccag gcactaagtg 1440ccacacatgt gtgtttactt
aaggtgatgt cagtatttat agggtgtgga gtgcaataaa 1500aatatgtgtt gactttaagt
gcgtggttta tgactcaggg gtggggactg tgggtatata 1560agcaggtgca gacctgtgtg
gtcagttcag agcaggactc atggagatct ggacggtctt 1620ggaagacttt caccagacta
gacagctgct agagaactca tcggcggaag tctcttacct 1680gtggagattc tgcttcggtg
ggcctctagc taagctagtc tatagggcca agcaggatta 1740taaggatcaa tttgaggata
ttttgagaga gtgtcctggt atttttgact ctctcaactt 1800gggccatcag tctcacttta
accagagtat tctgagagcc cttgactttt ccactcctgg 1860cagaactacc gccgcggtag
ccttttttgc ctttatcctt gacaaatgga gtcaagaaac 1920ccatttcagc agggattacc
gtctggactg cttagcagta gctttgtgga gaacatggag 1980gtgccagcgc ctgaatgcaa
tctccggcta cttgccagta cagccggtag acacgctgag 2040gatcctgagt ctccagtcac
cccaggaaca ccaacgccgc cagcagccgc agcaggagca 2100gcagcaagag gaggaggagg
accgagaaga gaacctgaga gccggtctgg accctccggt 2160ggcggaggag gaggagtagc
tgacttgttt cccgagctgc gccgggtgct gactaggtct 2220tccagtggac gggagagggg
gattaagcgg gagaggcatg aggagactag ccacagaact 2280gaactgactg tcagtctgat
gagccgcagg cgcccagaat cggtgtggtg gcatgaggtg 2340cagtcgcagg ggatagatga
ggtctcagtg atgcatgaga aatattccct agaacaagtc 2400aagacttgtt ggttggagcc
tgaggatgat tgggaggtag ccatcaggaa ttatgccaag 2460ctggctctga agccagacaa
gaagtacaag attaccaaac tgattaatat cagaaattcc 2520tgctacattt cagggaatgg
ggccgaggtg gagatcagta cccaggagag ggcggccttc 2580agatgttgta tgatgaatat
gtacccgggg gtggtgggca tggagggagt cacctttatg 2640aacacgaggt tcaggggtga
tgggtataat ggggtggtct ttatggccaa caccaagttg 2700acagtgcacg gatgctcctt
ctttggcttc aataacatgt gcatcgaggc ctggggcagt 2760gtttcagtga ggggatgcag
cttttcagcc aactggatgg gggtcgtggg cagaaccaag 2820agcgtggttt cagtgaagaa
atgcctgttt gagaggtgcc acctgggggt gatgagcgag 2880ggcgaagcca aagtcaaaca
ctgcgcctct accgagacgg gctgctttgt gctgatcaag 2940ggcaatgcca aagtcaagca
taacatgatc tgtggggcct cggatgagcg cggctaccag 3000atgctgacct gcgccggtgg
gaacagccat atgctggcca ccgtgcatgt ggcctcgcac 3060ccccgcaaga catggcccga
gttcgagcac aacgtcatga cccgctgcaa tgtgcacctg 3120gggtcccgcc gaggcatgtt
catgccctac cagtgcaaca tgcaatttgt gaaggtgctg 3180ctggagcccg atgccatgtc
cagagtgagc ctgacggggg tgtttgacat gaatgtggag 3240ctgtggaaaa ttctgagata
tgatgaatcc aagaccaggt gccgggcctg cgaatgcgga 3300ggcaagcacg ccaggcttca
gcccgtgtgt gtggaggtga cggaggacct gcgacccgat 3360catttggtgt tgtcctgcaa
cgggacggag ttcggctcca gcggggaaga atctgactag 3420agtgagtagt gtttgggggt
gggtgggagc ctgcatgatg ggcagaatga ctaaaatctg 3480tgtttttctg tgtgttgcag
cagcatgagc ggaagcgcct cctttgaggg aggggtattc 3540agcccttatc tgacggggcg
tctcccctcc tgggcgggag tgcgtcagaa tgtgatggga 3600tccacggtgg acggccggcc
cgtgcagccc gcgaactctt caaccctgac ctacgcgacc 3660ctgagctcct cgtccgtgga
cgcagctgcc gccgcagctg ctgcttccgc cgccagcgcc 3720gtgcgcggaa tggccctggg
cgccggctac tacagctctc tggtggccaa ctcgagttcc 3780accaataatc ccgccagcct
gaacgaggag aagctgttgc tgctgatggc ccagctcgag 3840gccctgaccc agcgcctggg
cgagctgacc cagcaggtgg ctcagctgca ggcggagacg 3900cgggccgcgg ttgccacggt
gaaaaccaaa taaaaaatga atcaataaat aaacggagac 3960ggttgttgat tttaacacag
agtcttgaat ctttatttga tttttcgcgc gcggtaggcc 4020ctggaccacc ggtctcgatc
attgagcacc cggtggatct tttccaggac ccggtagagg 4080tgggcttgga tgttgaggta
catgggcatg agcccgtccc gggggtggag gtagctccat 4140tgcagggcct cgtgctcggg
ggtggtgttg taaatcaccc agtcatagca ggggcgcagg 4200gcgtggtgct gcacgatgtc
tttgaggagg agactgatgg ccacgggcag ccccttggtg 4260taggtgttga cgaacctatt
gagctgggag ggatgcatgc ggggggagat gagatgcatc 4320ttggcctgga tcttgagatt
ggcgatgttc ccgcccagat cccgccgggg gttcatgttg 4380tgcaggacca ccagcacggt
gtatccggtg cacttgggga atttgtcatg caacttggaa 4440gggaaggcgt gaaagaattt
ggagacgccc ttgtgaccgc ccaggttttc catgcactca 4500tccatgatga tggcgatggg
cccgtgggcg gcggcctggg caaagacgtt tcgggggtcg 4560gacacatcgt agttgtggtc
ctgggtgagc tcgtcatagg ccattttaat gaatttgggg 4620cggagggtac ccgactgggg
gacaaaggtg ccctcgatcc cgggggcgta gttcccctcg 4680cagatctgca tctcccaggc
cttgagctcg gaggggggga tcatgtccac ctgcggggcg 4740atgaaaaaaa cggtttccgg
ggcgggggag atgagctgcg ccgaaagcag gttccggagc 4800agctgggact tgccgcagcc
ggtggggccg tagatgaccc cgatgaccgg ctgcaggtgg 4860tagttgaggg agagacagct
gccgtcctcg cggaggaggg gggccacctc gttcatcatc 4920tcgcgcacat gcatgttctc
gcgcacgagt tccgccagga ggcgctcgcc ccccagcgag 4980aggagctctt gcagcgaggc
gaagtttttc agcggcttga gcccgtcggc catgggcatt 5040ttggagaggg tctgttgcaa
gagttccaga cggtcccaga gctcggtgat gtgctctagg 5100gcatctcgat ccagcagacc
tcctcgtttc gcgggttggg gcgactgcgg gagtagggca 5160ccaggcgatg ggcgtccagc
gaggccaggg tccggtcctt ccagggtcgc agggtccgcg 5220tcagcgtggt ctccgtcacg
gtgaaggggt gcgcgccggg ctgggcgctt gcgagggtgc 5280gcttcaggct catccggctg
gtcgagaacc gctcccggtc ggcgccctgc gcgtcggcca 5340ggtagcaatt gagcatgagt
tcgtagttga gcgcctcggc cgcgtggccc ttggcgcgga 5400gcttaccttt ggaagtgtgt
ccgcagacgg gacagaggag ggacttgagg gcgtagagct 5460tgggggcgag gaagacggac
tcgggggcgt aggcgtccgc gccgcagctg gcgcagacgg 5520tctcgcactc cacgagccag
gtgaggtcgg ggcggtcggg gtcaaaaacg aggtttcctc 5580cgtgcttttt gatgcgtttc
ttacctctgg tctccatgag ctcgtgtccc cgctgggtga 5640caaagaggct gtccgtgtcc
ccgtagaccg actttatggg ccggtcctcg agcggggtgc 5700cgcggtcctc gtcgtagagg
aaccccgccc actccgagac gaaggcccgg gtccaggcca 5760gcacgaagga ggccacgtgg
gaggggtagc ggtcgttgtc caccagcggg tccaccttct 5820ccagggtatg caagcacatg
tccccctcgt ccacatccag gaaggtgatt ggcttgtaag 5880tgtaggccac gtgaccgggg
gtcccggccg ggggggtata aaagggggcg ggcccctgct 5940cgtcctcact gtcttccgga
tcgctgtcca ggagcgccag ctgttggggt aggtattccc 6000tctcgaaggc gggcatgacc
tcggcactca ggttgtcagt ttctagaaac gaggaggatt 6060tgatattgac ggtgccgttg
gagacgcctt tcatgagccc ctcgtccatc tggtcagaaa 6120agacgatctt tttgttgtcg
agcttggtgg cgaaggagcc gtagagggcg ttggagagca 6180gcttggcgat ggagcgcatg
gtctggttct tttccttgtc ggcgcgctcc ttggcggcga 6240tgttgagctg cacgtactcg
cgcgccacgc acttccattc ggggaagacg gtggtgagct 6300cgtcgggcac gattctgacc
cgccagccgc ggttgtgcag ggtgatgagg tccacgctgg 6360tggccacctc gccgcgcagg
ggctcgttgg tccagcagag gcgcccgccc ttgcgcgagc 6420agaagggggg cagcgggtcc
agcatgagct cgtcgggggg gtcggcgtcc acggtgaaga 6480tgccgggcag gagctcgggg
tcgaagtagc tgatgcaggt gcccagatcg tccagacttg 6540cttgccagtc gcgcacggcc
agcgcgcgct cgtaggggct gaggggcgtg ccccagggca 6600tggggtgcgt gagcgcggag
gcgtacatgc cgcagatgtc gtagacgtag aggggctcct 6660ggaggacgcc gatgtaggtg
gggtagcagc gccccccgcg gatgctggcg cgcacgtagt 6720cgtacagctc gtgcgagggc
gcgaggagcc ccgtgccgag attggagcgc tgcggctttt 6780cggcgcggta gacgatctgg
cggaagatgg cgtgggagtt ggaggagatg gtgggcctct 6840ggaagatgtt gaagtgggca
tggggcagtc cgaccgagtc cctgatgaag tgggcgtagg 6900agtcctgcag cttggcgacg
agctcggcgg tgacgaggac gtccagggcg cagtagtcga 6960gggtctcttg gatgatgtcg
tacttgagct ggcccttctg cttccacagc tcgcggttga 7020gaaggaactc ttcgcggtcc
ttccagtact cttcgagggg gaacccgtcc tgatcggcac 7080ggtaagagcc caccatgtag
aactggttga cggccttgta ggcgcagcag cccttctcca 7140cggggagggc gtaagcttgc
gcggccttgc gcagggaggt gtgggtgagg gcgaaggtgt 7200cgcgcaccat gactttgagg
aactggtgct tgaagtcgag gtcgtcgcag ccgccctgct 7260cccagagctg gaagtccgtg
cgcttcttgt aggcggggtt gggcaaagcg aaagtaacat 7320cgttgaagag gatcttgccc
gcgcggggca tgaagttgcg agtgatgcgg aaaggctggg 7380gcacctcggc ccggttgttg
atgacctggg cggcgaggac gatctcgtcg aagccgttga 7440tgttgtgccc gacgatgtag
agttccacga atcgcgggcg gcccttgacg tggggcagct 7500tcttgagctc gtcgtaggtg
agctcggcgg ggtcgctgag cccgtgctgc tcgagggccc 7560agtcggcgac gtgggggttg
gcgctgagga aggaagtcca gagatccacg gccagggcgg 7620tctgcaagcg gtcccggtac
tgacggaact gctggcccac ggccattttt tcgggggtga 7680cgcagtagaa ggtgcggggg
tcgccgtgcc agcggtccca cttgagctgg agggcgaggt 7740cgtgggcgag ctcgacgagc
ggcgggtccc cggagagttt catgaccagc atgaagggga 7800cgagctgctt gccgaaggac
cccatccagg tgtaggtttc cacatcgtag gtgaggaaga 7860gcctttcggt gcgaggatgc
gagccgatgg ggaagaactg gatctcctgc caccagttgg 7920aggaatggct gttgatgtga
tggaagtaga aatgccgacg gcgcgccgag cactcgtgct 7980tgtgtttata caagcgtccg
cagtgctcgc aacgctgcac gggatgcacg tgctgcacga 8040gctgtacctg ggttcctttg
acgaggaatt tcagtgggca gtggagcgct ggcggctgca 8100tctggtgctg tactacgtcc
tggccatcgg cgtggccatc gtctgcctcg atggtggtca 8160tgctgacgag cccgcgcggg
aggcaggtcc agacctcggc tcggacgggt cggagagcga 8220ggacgagggc gcgcaggccg
gagctgtcca gggtcctgag acgctgcgga gtcaggtcag 8280tgggcagcgg cggcgcgcgg
ttgacttgca ggagcttttc cagggcgcgc gggaggtcca 8340gatggtactt gatctccacg
gcgccgttgg tggcgacgtc cacggcttgc agggtcccgt 8400gcccctgggg cgccaccacc
gtgccccgtt tcttcttggg cggcggcggc tccatgctta 8460gaagcggcgg cgaggacgcg
cgccgggcgg caggggcggc tcggggcccg gaggcagggg 8520cggcaggggc acgtcggcgc
cgcgcgcggg caggttctgg tactgcgccc ggagaagact 8580ggcgtgagcg acgacgcgac
ggttgacgtc ctggatctga cgcctctggg tgaaggccac 8640gggacccgtg agtttgaacc
tgaaagagag ttcgacagaa tcaatctcgg tatcgttgac 8700ggcggcctgc cgcaggatct
cttgcacgtc gcccgagttg tcctggtagg cgatctcggt 8760catgaactgc tcgatctcct
cctcctgaag gtctccgcgg ccggcgcgct cgacggtggc 8820cgcgaggtcg ttggagatgc
ggcccatgag ctgcgagaag gcgttcatgc cggcctcgtt 8880ccagacgcgg ctgtagacca
cggctccgtc ggggtcgcgc gcgcgcatga ccacctgggc 8940gaggttgagc tcgacgtggc
gcgtgaagac cgcgtagttg cagaggcgct ggtagaggta 9000gttgagcgtg gtggcgatgt
gctcggtgac gaagaagtac atgatccagc ggcggagcgg 9060catctcgctg acgtcgccca
gggcttccaa gcgctccatg gcctcgtaga agtccacggc 9120gaagttgaaa aactgggagt
tgcgcgccga gacggtcaac tcctcctcca gaagacggat 9180gagctcggcg atggtggcgc
gcacctcgcg ctcgaaggcc ccggggggct cctcttccat 9240ttcctcctct tcctcctcca
ctaacatctc ttctacttcc tcctcaggag gcggcggcgg 9300gggaggggcc ctgcgtcgcc
ggcggcgcac gggcagacgg tcgatgaagc gctcgatggt 9360ctccccgcgc cggcgacgca
tggtctcggt gacggcgcgc ccgtcctcgc ggggccgcag 9420cgtgaagacg ccgccgcgca
tctccaggtg gccgccgggg gggtctccgt tgggcaggga 9480gagggcgctg acgatgcatc
ttatcaattg acccgtaggg actccgcgca aggacctgag 9540cgtctcgaga tccacgggat
ccgaaaaccg ctgaacgaag gcttcgagcc agtcgcagtc 9600gcaaggtagg ctgagcccgg
tttcttgttc ttcgggtatt tggtcgggag gcgggcgggc 9660gatgctgctg gtgatgaagt
tgaagtaggc ggtcctgaga cggcggatgg tggcgaggag 9720caccaggtcc ttgggcccgg
cttgctggat gcgcagacgg tcggccatgc cccaggcgtg 9780gtcctgacac ctggcgaggt
ccttgtagta gtcctgcatg agccgctcca cgggcacctc 9840ctcctcgccc gcgcggccgt
gcatgcgcgt gagcccgaac ccgcgctgcg gctggacgag 9900cgccaggtcg gcgacgacgc
gctcggcgag gatggcctgc tggatctggg tgagggtggt 9960ctggaagtcg tcgaagtcga
cgaagcggtg gtaggctccg gtgttgatgg tgtaggagca 10020gttggccatg acggaccagt
tgacggtctg gtggccgggg cgcacgagct cgtggtactt 10080gaggcgcgag taggcgcgcg
tgtcgaagat gtagtcgttg caggtgcgca cgaggtactg 10140gtatccgacg aggaagtgcg
gcggcggctg gcggtagagc ggccatcgct cggtggcggg 10200ggcgccgggc gcgaggtcct
cgagcatgag gcggtggtag ccgtagatgt acctggacat 10260ccaggtgatg ccggcggcgg
tggtggaggc gcgcgggaac tcgcggacgc ggttccagat 10320gttgcgcagc ggcaggaagt
agttcatggt ggccgcggtc tggcccgtga ggcgcgcgca 10380gtcgtggatg ctctagacat
acgggcaaaa acgaaagcgg tcagcggctc gactccgtgg 10440cctggaggct aagcgaacgg
gttgggctgc gcgtgtaccc cggttcgaat ctcgaatcag 10500gctggagccg cagctaacgt
ggtactggca ctcccgtctc gacccaagcc tgctaacgaa 10560acctccagga tacggaggcg
ggtcgttttt tggccttggt cgctggtcat gaaaaactag 10620taagcgcgga aagcggccgc
ccgcgatggc tcgctgccgt agtctggaga aagaatcgcc 10680agggttgcgt tgcggtgtgc
cccggttcga gcctcagcgc tcggcgccgg ccggattccg 10740cggctaacgt gggcgtggct
gccccgtcgt ttccaagacc ccttagccag ccgacttctc 10800cagttacgga gcgagcccct
ctttttttct tgtgtttttg ccagatgcat cccgtactgc 10860ggcagatgcg cccccaccct
ccaccacaac cgcccctacc gcagcagcag caacagccgg 10920cgcttctgcc cccgccccag
cagcagcagc cagccactac cgcggcggcc gccgtgagcg 10980gagccggcgt tcagtatgac
ctggccttgg aagagggcga ggggctggcg cggctggggg 11040cgtcgtcgcc ggagcggcac
ccgcgcgtgc agatgaaaag ggacgctcgc gaggcctacg 11100tgcccaagca gaacctgttc
agagacagga gcggcgagga gcccgaggag atgcgcgcct 11160cccgcttcca cgcggggcgg
gagctgcggc gcggcctgga ccgaaagcgg gtgctgaggg 11220acgaggattt cgaggcggac
gagctgacgg ggatcagccc cgcgcgcgcg cacgtggccg 11280cggccaacct ggtcacggcg
tacgagcaga ccgtgaagga ggagagcaac ttccaaaaat 11340ccttcaacaa ccacgtgcgc
acgctgatcg cgcgcgagga ggtgaccctg ggcctgatgc 11400acctgtggga cctgctggag
gccatcgtgc agaaccccac gagcaagccg ctgacggcgc 11460agctgtttct ggtggtgcag
cacagtcggg acaacgagac gttcagggag gcgctgctga 11520atatcaccga gcccgagggc
cgctggctcc tggacctggt gaacattctg cagagcatcg 11580tggtgcagga gcgcgggctg
ccgctgtccg agaagctggc ggccatcaac ttctcggtgc 11640tgagcctggg caagtactac
gctaggaaga tctacaagac cccgtacgtg cccatagaca 11700aggaggtgaa gatcgatggg
ttttacatgc gcatgaccct gaaagtgctg accctgagcg 11760acgatctggg ggtgtaccgc
aacgacagga tgcaccgcgc ggtgagcgcc agccgccggc 11820gcgagctgag cgaccaggag
ctgatgcaca gcctgcagcg ggccctgacc ggggccggga 11880ccgaggggga gagctacttt
gacatgggcg cggacctgcg ctggcagccc agccgccggg 11940ccttggaagc tgccggcggc
gtgccctacg tggaggaggt ggacgatgag gaggaggagg 12000gcgagtacct ggaagactga
tggcgcgacc gtatttttgc tagatgcagc aacagccacc 12060gccgccgcct cctgatcccg
cgatgcgggc ggcgctgcag agccagccgt ccggcattaa 12120ctcctcggac gattggaccc
aggccatgca acgcatcatg gcgctgacga cccgcaatcc 12180cgaagccttt agacagcagc
ctcaggccaa ccggctctcg gccatcctgg aggccgtggt 12240gccctcgcgc tcgaacccca
cgcacgagaa ggtgctggcc atcgtgaacg cgctggtgga 12300gaacaaggcc atccgcggcg
acgaggccgg gctggtgtac aacgcgctgc tggagcgcgt 12360ggcccgctac aacagcacca
acgtgcagac gaacctggac cgcatggtga ccgacgtgcg 12420cgaggcggtg tcgcagcgcg
agcggttcca ccgcgagtcg aacctgggct ccatggtggc 12480gctgaacgcc ttcctgagca
cgcagcccgc caacgtgccc cggggccagg aggactacac 12540caacttcatc agcgcgctgc
ggctgatggt ggccgaggtg ccccagagcg aggtgtacca 12600gtcggggccg gactacttct
tccagaccag tcgccagggc ttgcagaccg tgaacctgag 12660ccaggctttc aagaacttgc
agggactgtg gggcgtgcag gccccggtcg gggaccgcgc 12720gacggtgtcg agcctgctga
cgccgaactc gcgcctgctg ctgctgctgg tggcgccctt 12780cacggacagc ggcagcgtga
gccgcgactc gtacctgggc tacctgctta acctgtaccg 12840cgaggccatc gggcaggcgc
acgtggacga gcagacctac caggagatca cccacgtgag 12900ccgcgcgctg ggccaggagg
acccgggcaa cctggaggcc accctgaact tcctgctgac 12960caaccggtcg cagaagatcc
cgccccagta cgcgctgagc accgaggagg agcgcatcct 13020gcgctacgtg cagcagagcg
tggggctgtt cttgatgcag gagggggcca cgcccagcgc 13080cgcgctcgac atgaccgcgc
gcaacatgga gcccagcatg tacgcccgca accgcccgtt 13140catcaataag ctgatggact
acttgcatcg ggcggccgcc atgaactcgg actactttac 13200caacgccatc ttgaacccgc
actggctccc gccgcccggg ttctacacgg gcgagtacga 13260catgcccgac cccaacgacg
ggttcctgtg ggacgacgtg gacagcagcg tgttctcgcc 13320gcggcccacc accaccaccg
tgtggaagaa agagggcggg gaccggcggc cgtcctcggc 13380gctgtccggt cgcgcgggtg
ctgccgcggc ggtgcccgag gctgccagcc ccttcccgag 13440cctgcccttt tcgctgaaca
gcgtgcgcag cagcgagctg ggtcggctga cgcggccgcg 13500cctgctgggc gaggaggagt
acctgaacga ctccttgttg aagcccgagc gcgagaagaa 13560cttccccaat aacgggatag
agagcctggt ggacaagatg agccgctgga agacgtacgc 13620gcacgagcac agggacgagc
cccgagctag cagcgcaggc acccgtagac gccagcggca 13680cgacaggcag cggggactgg
tgtgggacga tgaggattcc gccgacgaca gcagcgtgtt 13740ggacttgggt gggagtggtg
gtggtaaccc gttcgctcac ctgcgccccc gtatcgggcg 13800cctgatgtaa gaatctgaaa
aaataaaaga cggtactcac caaggccatg gcgaccagcg 13860tgcgttcttc tctgttgttt
gtagtagtat gatgaggcgc gtgtacccgg agggtcctcc 13920tccctcgtac gagagcgtga
tgcagcaggc ggtggcggcg gcgatgcagc ccccgctgga 13980ggcgccttac gtgcccccgc
ggtacctggc gcctacggag gggcggaaca gcattcgtta 14040ctcggagctg gcacccttgt
acgataccac ccggttgtac ctggtggaca acaagtcggc 14100ggacatcgcc tcgctgaact
accagaacga ccacagcaac ttcctgacca ccgtggtgca 14160gaacaacgat ttcaccccca
cggaggccag cacccagacc atcaactttg acgagcgctc 14220gcggtggggc ggccagctga
aaaccatcat gcacaccaac atgcccaacg tgaacgagtt 14280catgtacagc aacaagttca
aggcgcgggt gatggtctcg cgcaagaccc ccaacggggt 14340cacggtaggg gatgattatg
atggtagtca ggacgagctg acctacgagt gggtggagtt 14400tgagctgccc gagggcaact
tctcggtgac catgaccatc gatctgatga acaacgccat 14460catcgacaac tacttggcgg
tggggcggca gaacggggtg ctggagagcg acatcggcgt 14520gaagttcgac acgcgcaact
tccggctggg ctgggacccc gtgaccgagc tggtgatgcc 14580gggcgtgtac accaacgagg
ccttccaccc cgacatcgtc ctgctgcccg gctgcggcgt 14640ggacttcacc gagagccgcc
tcagcaacct gctgggcatc cgcaagcggc agcccttcca 14700ggagggcttc cagatcctgt
acgaggacct ggaggggggc aacatccccg cgctcttgga 14760tgtcgaagcc tatgaagaaa
gtaaggaaaa agcagaggct gaggcaacta cagccgtggc 14820taccgccgcg actgtggcag
atgccactgt caccaggggc gatacattcg ccacccaggc 14880ggaggaagca gccgccctag
cggcgaccga tgatagtgaa agtaagatag tcatcaagcc 14940ggtggagaag gacagcaaga
acaggagcta caacgttcta ccggatggaa agaacaccgc 15000ctaccgcagc tggtacctgg
cctacaacta cggcgacccc gagaagggcg tgcgctcctg 15060gacgctgctc accacctcgg
acgtcacctg cggcgtggag caagtctact ggtcgctgcc 15120cgacatgatg caagacccgg
tcaccttccg ctccacgcga caagttagca actacccggt 15180ggtgggcgcc gagctcctgc
ccgtctactc caagagcttc ttcaacgagc aggccgtcta 15240ctcgcagcag ctgcgtgcct
tcacctcgct cacgcacgtc ttcaaccgct tccccgagaa 15300ccagatcctc gtccgcccgc
ccgcgcccac cattaccacc gtcagtgaaa acgttcctgc 15360tctcacagat cacgggaccc
tgccgctgcg cagcagtatc cggggagtcc agcgcgtgac 15420cgtcactgac gccagacgcc
gcacctgccc ctacgtctac aaggccctgg gcgtagtcgc 15480gccgcgcgtc ctctcgagcc
gcaccttcta aaaaatgtcc attctcatct cgcccagtaa 15540taacaccggt tggggcctgc
gcgcgcccag caagatgtac ggaggcgctc gccaacgctc 15600cacgcaacac cccgtgcgcg
tgcgcgggca cttccgcgct ccctggggcg ccctcaaggg 15660ccgcgtgcgc tcgcgcacca
ccgtcgacga cgtgatcgac caggtggtgg ccgacgcgcg 15720caactacacg cccgccgccg
cgcccgcctc caccgtggac gccgtcatcg acagcgtggt 15780ggccgacgcg cgccggtacg
cccgcgccaa gagccggcgg cggcgcatcg cccggcggca 15840ccggagcacc cccgccatgc
gcgcggcgcg agccttgctg cgcagggcca ggcgcacggg 15900acgcagggcc atgctcaggg
cggccagacg cgcggcctcc ggcagcagca gcgccggcag 15960gacccgcaga cgcgcggcca
cggcggcggc ggcggccatc gccagcatgt cccgcccgcg 16020gcgcggcaac gtgtactggg
tgcgcgacgc cgccaccggt gtgcgcgtgc ccgtgcgcac 16080ccgcccccct cgcacttgaa
gatgctgact tcgcgatgtt gatgtgtccc agcggcgagg 16140aggatgtcca agcgcaaata
caaggaagag atgctccagg tcatcgcgcc tgagatctac 16200ggccccgcgg cggcggtgaa
ggaggaaaga aagccccgca aactgaagcg ggtcaaaaag 16260gacaaaaagg aggaggaaga
tgacggactg gtggagtttg tgcgcgagtt cgccccccgg 16320cggcgcgtgc agtggcgcgg
gcggaaagtg aaaccggtgc tgcggcccgg caccacggtg 16380gtcttcacgc ccggcgagcg
ttccggctcc gcctccaagc gctcctacga cgaggtgtac 16440ggggacgagg acatcctcga
gcaggcggtc gagcgtctgg gcgagtttgc ttacggcaag 16500cgcagccgcc ccgcgccctt
gaaagaggag gcggtgtcca tcccgctgga ccacggcaac 16560cccacgccga gcctgaagcc
ggtgaccctg cagcaggtgc tgccgagcgc ggcgccgcgc 16620cggggcttca agcgcgaggg
cggcgaggat ctgtacccga ccatgcagct gatggtgccc 16680aagcgccaga agctggagga
cgtgctggag cacatgaagg tggaccccga ggtgcagccc 16740gaggtcaagg tgcggcccat
caagcaggtg gccccgggcc tgggcgtgca gaccgtggac 16800atcaagatcc ccacggagcc
catggaaacg cagaccgagc ccgtgaagcc cagcaccagc 16860accatggagg tgcagacgga
tccctggatg ccagcggctt ccaccaccac cactcgccga 16920agacgcaagt acggcgcggc
cagcctgctg atgcccaact acgcgctgca tccttccatc 16980atccccacgc cgggctaccg
cggcacgcgc ttctaccgcg gctacaccag cagccgccgc 17040cgcaagacca ccacccgccg
ccgtcgtcgc agccgccgca gcagcaccgc gacttccgcc 17100ttggtgcgga gagtgtatcg
cagcgggcgc gagcctctga ccctgccgcg cgcgcgctac 17160cacccgagca tcgccattta
actaccgcct cctacttgca gatatggccc tcacatgccg 17220cctccgcgtc cccattacgg
gctaccgagg aagaaagccg cgccgtagaa ggctgacggg 17280gaacgggctg cgtcgccatc
accaccggcg gcggcgcgcc atcagcaagc ggttgggggg 17340aggcttcctg cccgcgctga
tccccatcat cgccgcggcg atcggggcga tccccggcat 17400agcttccgtg gcggtgcagg
cctctcagcg ccactgagac acaaaaaagc atggatttgt 17460aataaaaaaa tggactgacg
ctcctggtcc tgtgatgtgt gtttttagat ggaagacatc 17520aatttttcgt ccctggcacc
gcgacacggc acgcggccgt ttatgggcac ctggagcgac 17580atcggcaaca gccaactgaa
cgggggcgcc ttcaattgga gcagtctctg gagcgggctt 17640aagaatttcg ggtccacgct
caaaacctat ggcaacaagg cgtggaacag cagcacaggg 17700caggcgctga gggaaaagct
gaaagagcag aacttccagc agaaggtggt cgatggcctg 17760gcctcgggca tcaacggggt
ggtggacctg gccaaccagg ccgtgcagaa acagatcaac 17820agccgcctgg acgcggtccc
gcccgcgggg tccgtggaga tgccccaggt ggaggaggag 17880ctgcctcccc tggacaagcg
cggcgacaag cgaccgcgtc ccgacgcgga ggagacgctg 17940ctgacgcaca cggacgagcc
gcccccgtac gaggaggcgg tgaaactggg tctgcccacc 18000acgcggcccg tggcgcctct
ggccaccggg gtgctgaaac ccagcagcag cagccagccc 18060gcgaccctgg acttgcctcc
gcctgcttcc cgcccctcca cagtggctaa gcccctgccg 18120ccggtggccg tcgcgtcgcg
cgccccccga ggccgccccc aggcgaactg gcagagcact 18180ctgaacagca tcgtgggtct
gggagtgcag agtgtgaagc gccgccgctg ctattaaaag 18240acactgtagc gcttaacttg
cttgtctgtg tgtgtatatg tatgtccgcc gaccagaagg 18300aggaagaggc gcgtcgccga
gttgcaagat ggccacccca tcgatgctgc cccagtgggc 18360gtacatgcac atcgccggac
aggacgcttc ggagtacctg agtccgggtc tggtgcagtt 18420cgcccgcgcc acagacacct
acttcagtct ggggaacaag tttaggaacc ccacggtggc 18480gcccacgcac gatgtgacca
ccgaccgcag ccagcggctg acgctgcgct tcgtgcccgt 18540ggaccgcgag gacaacacct
actcgtacaa agtgcgctac acgctggccg tgggcgacaa 18600ccgcgtgctg gacatggcca
gcacctactt tgacatccgc ggcgtgctgg atcggggccc 18660cagcttcaaa ccctactccg
gcaccgccta caacagccta gctcccaagg gagcgcccaa 18720cacctcacag tggaaggatt
ccgacagcaa aatgcatact tttggagttg ctgccatgcc 18780cggtgttgtt ggtaaaaaaa
tagaagccga tggtctgcct attggaatag attcatcctc 18840tggaactgac accataattt
atgctgataa aactttccaa ccagagccac aggttggaag 18900tgacagttgg gtcgacacca
atggtgcaga ggaaaaatat ggaggtagag ctcttaagga 18960cactacaaac atgaagccct
gctacggttc ttttgccagg cctaccaaca aagaaggtgg 19020acaggctaac ataaaagatt
ctgaaactgc cagcactact cctaactatg atatagattt 19080ggcattcttt gacagcaaaa
atattgcagc taactacgat ccagatattg taatgtacac 19140agaaaatgtt gagttgcaaa
ctccagatac tcatattgtg tttaagccag gaacttcaga 19200tgaaagttca gaagccaatt
tgggccagca ggccatgccc aacagaccca actacatcgg 19260gttcagagac aactttatcg
ggctcatgta ctacaacagc actggcaata tgggtgtact 19320ggctggtcag gcctcccagc
taaatgctgt ggtggacttg caggacagaa acaccgaact 19380gtcctaccag ctcttgcttg
actctctggg tgacagaacc aggtatttca gtatgtggaa 19440tcaggcggtg gacagctatg
accccgatgt gcgcattatt gaaaatcacg gtgtggagga 19500tgaactcccc aattattgct
tccctttgaa tggtgtaggc tttacagata cttaccaggg 19560tgttaaagtt aagacagata
cagccgctac tggtaccaat ggaacgcagt gggacaaaga 19620tgataccaca gtcagcactg
ccaatgagat ccactcaggc aatcctttcg ccatggagat 19680caacatccag gccaacctgt
ggcggaactt cctctacgcg aacgtggcgc tgtacctgcc 19740cgactcctac aagtacacgc
cggccaacat cacgctgccg accaacacca acacctacga 19800ttacatgaac ggccgcgtgg
tggcgccctc gctggtggac gcctacatca acatcggggc 19860gcgctggtcg ctggacccca
tggacaacgt caaccccttc aaccaccacc gcaacgcggg 19920cctgcgctac cgctccatgc
tcctgggcaa cgggcgctac gtgcccttcc acatccaggt 19980gccccaaaag tttttcgcca
tcaagagcct cctgctcctg cccgggtcct acacctacga 20040gtggaacttc cgcaaggacg
tcaacatgat cctgcagagc tccctcggca acgacctgcg 20100cacggacggg gcctccatcg
ccttcaccag catcaacctc tacgccacct tcttccccat 20160ggcgcacaac accgcctcca
cgctcgaggc catgctgcgc aacgacacca acgaccagtc 20220cttcaacgac tacctctcgg
cggccaacat gctctacccc atcccggcca acgccaccaa 20280cgtgcccatc tccatcccct
cgcgcaactg ggccgccttc cgcggatggt ccttcacgcg 20340cctcaagacc cgcgagacgc
cctcgctcgg ctccgggttc gacccctact tcgtctactc 20400gggctccatc ccctacctcg
acggcacctt ctacctcaac cacaccttca agaaggtctc 20460catcaccttc gactcctccg
tcagctggcc cggcaacgac cgcctcctga cgcccaacga 20520gttcgaaatc aagcgcaccg
tcgacggaga gggatacaac gtggcccagt gcaacatgac 20580caaggactgg ttcctggtcc
agatgctggc ccactacaac atcggctacc agggcttcta 20640cgtgcccgag ggctacaagg
accgcatgta ctccttcttc cgcaacttcc agcccatgag 20700ccgccaggtc gtggacgagg
tcaactacaa ggactaccag gccgtcaccc tggcctacca 20760gcacaacaac tcgggcttcg
tcggctacct cgcgcccacc atgcgccagg gccagcccta 20820ccccgccaac tacccctacc
cgctcatcgg caagagcgcc gtcgccagcg tcacccagaa 20880aaagttcctc tgcgaccggg
tcatgtggcg catccccttc tccagcaact tcatgtccat 20940gggcgcgctc accgacctcg
gccagaacat gctctacgcc aactccgccc acgcgctaga 21000catgaatttc gaagtcgacc
ccatggatga gtccaccctt ctctatgttg tcttcgaagt 21060cttcgacgtc gtccgagtgc
accagcccca ccgcggcgtc atcgaggccg tctacctgcg 21120cacgcccttc tcggccggca
acgccaccac ctaaagcccc gctcttgctt cttgcaagat 21180gacggcctgt ggctccggcg
agcaggagct cagggccatc ctccgcgacc tgggctgcgg 21240gccctgcttc ctgggcacct
tcgacaagcg cttcccggga ttcatggccc cgcacaagct 21300ggcctgcgcc atcgtcaaca
cggccggccg cgagaccggg ggcgagcact ggctggcctt 21360cgcctggaac ccgcgctccc
acacctgcta cctcttcgac cccttcgggt tctcggacga 21420gcgcctcaag cagatctacc
agttcgagta cgagggcctg ctgcgccgca gcgccctggc 21480caccgaggac cgctgcatca
ccctggaaaa gtccacccag accgtgcagg gtccgcgctc 21540ggccgcctgc gggctcttct
gctgcatgtt cctgcacgcc ttcgtgcact ggcccgaccg 21600ccccatggac aagaacccca
ccatgaactt gctgacgggg gtgcccaacg gcatgctcca 21660gtcgccccag gtggaaccca
ccctgcgccg caaccaggag gcgctctacc gcttcctcaa 21720cgcccactcc gcctactttc
gctcccaccg cgcgcgcatc gagaaggcca ccgccttcga 21780ccgcatgaat caagacatgt
aaactgtgtg tatgtgaatg ctttattcat cataataaac 21840agcacatgtt tatgccacct
tctctgaggc tctgacttta tttagaaatc gaaggggttc 21900tgccggctct cggcgtgccc
cgcgggcagg gatacgttgc ggaactggta cttgggcagc 21960cacttgaact cggggatcag
cagcttcggc acggggaggt cggggaacga gtcgctccac 22020agcttgcgcg tgagttgcag
ggcgcccagc aggtcgggcg cggagatctt gaaatcgcag 22080ttgggacccg cgttctgcgc
gcgagagttg cggtacacgg ggttgcagca ctggaacacc 22140atcagggccg ggtgcttcac
gctcgccagc accgtcgcgt cggtgatgcc ctccacgtcc 22200agatcctcgg cgttggccat
cccgaagggg gtcatcttgc aggtctgccg ccccatgctg 22260ggcacgcagc cgggcttgtg
gttgcaatcg cagtgcaggg ggatcagcat catctgagcc 22320tgctcggagc tcatgcccgg
gtacatggcc ttcatgaaag cctccagctg gcggaaggcc 22380tgctgcgcct tgccgccctc
ggtgaagaag accccacagg acttgctaga gaactggttg 22440gtggcgcagc ccgcgtcgtg
cacgcagcag cgcgcgtcgt tgttggccag ctgcaccacg 22500ctgcgccccc agcggttctg
ggtgatcttg gcccggtcgg ggttctcctt cagcgcgcgc 22560tgcccgttct cgctcgccac
atccatctcg atcgtgtgct ccttctggat catcacggtc 22620ccgtgcaggc accgcagctt
gccctcggcc tcggtgcacc cgtgcagcca cagcgcgcag 22680ccggtgcact cccagttctt
gtgggcgatc tgggagtgcg agtgcacgaa gccctgcagg 22740aagcggccca tcatcgtggt
cagggtcttg ttgctggtga aggtcagcgg gatgccgcgg 22800tgctcctcgt tcacatacag
gtggcagatg cggcggtaca cctcgccctg ctcgggcatc 22860agctggaagg cggacttcag
gtcgctctcc acgcggtacc gctccatcag cagcgtcatc 22920acttccatgc ccttctccca
ggccgaaacg atcggcaggc tcagggggtt cttcaccgtc 22980atcttagtcg ccgccgccga
agtcaggggg tcgttctcgt ccagggtctc aaacactcgc 23040ttgccgtcct tctcggtgat
gcgcacgggg ggaaagctga agcccacggc cgccagctcc 23100tcctcggcct gcctttcgtc
ctcgctgtcc tggctgatgt cttgcaaagg cacatgcttg 23160gtcttgcggg gtttcttttt
gggcggcaga ggcggcggcg gagacgtgct gggcgagcgc 23220gagttctcgc tcaccacgac
tatttcttct tcttggccgt cgtccgagac cacgcggcgg 23280taggcatgcc tcttctgggg
cagaggcgga ggcgacgggc tctcgcggtt cggcgggcgg 23340ctggcagagc cccttccgcg
ttcgggggtg cgctcctggc ggcgctgctc tgactgactt 23400cctccgcggc cggccattgt
gttctcctag ggagcaacaa gcatggagac tcagccatcg 23460tcgccaacat cgccatctgc
ccccgccgcc gacgagaacc agcagcagca gaatgaaagc 23520ttaaccgccc cgccgcccag
ccccacctcc gacgccgccg cggccccaga catgcaagag 23580atggaggaat ccatcgagat
tgacctgggc tacgtgacgc ccgcggagca cgaggaggag 23640ctggcagcgc gcttttcagc
cccggaagag aaccaccaag agcagccaga gcaggaagca 23700gagagcgagc agcagcaggc
tgggctcgag catggcgact acctgagcgg ggcagaggac 23760gtgctcatca agcatctggc
ccgccaaagc atcatcgtca aggacgcgct gctcgaccgc 23820gccgaggtgc ccctcagcgt
ggcggagctc agccgcgcct acgagcgcaa cctcttctcg 23880ccgcgcgtgc cccccaagcg
ccagcccaac ggcacctgcg agcccaaccc gcgcctcaac 23940ttctacccgg tcttcgcggt
gcccgaggcc ctggccacct accacctctt tttcaagaac 24000caaaggatcc ccgtctcctg
ccgcgccaac cgcacccgcg ccgacgccct gctcaacctg 24060ggtcccggcg cccgcctacc
tgatatcacc tccttggaag aggttcccaa gatcttcgag 24120ggtctgggca gcgacgagac
tcgggccgcg aacgctctgc aaggaagcgg agaggagcat 24180gagcaccaca gcgccctggt
ggagttggaa ggcgacaacg cgcgcctggc ggtgctcaag 24240cgcacggtcg agctgaccca
cttcgcctac ccggcgctca acctgccccc caaggtcatg 24300agcgccgtca tggaccaggt
gctcatcaag cgcgcctcgc ccctctcaga ggaggagatg 24360caggaccccg agagctcgga
cgagggcaag cccgtggtca gcgacgagca gctggcgcgc 24420tggctgggag cgagcagcac
cccccagagc ctggaagagc ggcgcaagct catgatggcc 24480gtggtcctgg tgaccgtgga
gctggagtgt ctgcgccgct tcttcgccga cgcggagacc 24540ctgcgcaagg tcgaggagaa
cctgcactac ctcttcaggc acgggttcgt gcgccaggcc 24600tgcaagatct ccaacgtgga
gctgaccaac ctggtctcct acatgggcat cctgcacgag 24660aaccgcctgg ggcagaacgt
gctgcacacc accctgcgcg gggaggcccg ccgcgactac 24720atccgcgact gcgtctacct
gtacctctgc cacacctggc agacgggcat gggcgtgtgg 24780cagcagtgcc tggaggagca
gaacctgaaa gagctctgca agctcctgca gaagaacctc 24840aaggccctgt ggaccgggtt
cgacgagcgc accaccgcct cggacctggc cgacctcatc 24900ttccccgagc gcctgcggct
gacgctgcgc aacgggctgc ccgactttat gagccaaagc 24960atgttgcaaa actttcgctc
tttcatcctc gaacgctccg ggatcctgcc cgccacctgc 25020tccgcactgc cctcggactt
cgtgccgctg accttccgcg agtgcccccc gccgctctgg 25080agccactgct acttgctgcg
cctggccaac tacctggcct accactcgga cgtgatcgag 25140gacgtcagca gcgagggtct
gctcgagtgc cactgccgct gcaacctctg cacgccgcac 25200cgctccttgg cctgcaaccc
ccagctgctg agcgagaccc agatcatcgg caccttcgag 25260ttgcaaggcc ccggcgaggg
caaggggggt ctcaaactca ccccggggct gtggacctcg 25320gcctacttgc gcaagttcgt
gcccgaggac taccatccct tcgagatcag gttctacgag 25380gaccaatccc agccgcccaa
ggccgagctg tcggcctgcg tcatcaccca gggggccatc 25440ctggcccaat tgcaagccat
ccagaaatcc cgccaagaat ttctgctgaa aaagggccac 25500ggggtctact tggaccccca
gaccggagag gagctcaacc ccagcttccc ccaggatgcc 25560ccgaggaagc agcaagaagc
tgaaagtgga gctgccgctg ccgccggagg atttggagga 25620agactgggag agcagtcagg
cagaggagat ggaagactgg gacagcactc aggcagagga 25680ggacagcctg caagacagtc
tggaggagga agacgaggtg gaggaggagg cagaggaaga 25740agcagccgcc gccagaccgt
cgtcctcggc ggaggagaaa gcaagcagca cggataccat 25800ctccgctccg ggtcggggtc
gcggcggccg ggcccacagt agatgggacg agaccgggcg 25860cttcccgaac cccaccaccc
agaccggtaa gaaggagcgg cagggataca agtcctggcg 25920ggggcacaaa aacgccatcg
tctcctgctt gcaagcctgc gggggcaaca tctccttcac 25980ccggcgctac ctgctcttcc
accgcggggt gaacttcccc cgcaacatct tgcattacta 26040ccgtcacctc cacagcccct
actactgttt ccaagaagag gcagaaaccc agcagcagca 26100gcagaaaacc agcggcagca
gcagcagcta gaaaatccac agcggcggca ggtggactga 26160ggatcgcggc gaacgagccg
gcgcagaccc gggagctgag gaaccggatc tttcccaccc 26220tctatgccat cttccagcag
agtcgggggc aggagcagga actgaaagtc aagaaccgtt 26280ctctgcgctc gctcacccgc
agttgtctgt atcacaagag cgaagaccaa cttcagcgca 26340ctctcgagga cgccgaggct
ctcttcaaca agtactgcgc gctcactctt aaagagtagc 26400ccgcgcccgc ccacacacgg
aaaaaggcgg gaattacgtc accacctgcg cccttcgccc 26460gaccatcatc atgagcaaag
agattcccac gccttacatg tggagctacc agccccagat 26520gggcctggcc gccggcgccg
cccaggacta ctccacccgc atgaactggc tcagtgccgg 26580gcccgcgatg atctcacggg
tgaatgacat ccgcgcccac cgaaaccaga tactcctaga 26640acagtcagcg atcaccgcca
cgccccgcca tcaccttaat ccgcgtaatt ggcccgccgc 26700cctggtgtac caggaaattc
cccagcccac gaccgtacta cttccgcgag acgcccaggc 26760cgaagtccag ctgactaact
caggtgtcca gctggccggc ggcgccgccc tgtgtcgtca 26820ccgccccgct cagggtataa
agcggctggt gatccgaggc agaggcacac agctcaacga 26880cgaggtggtg agctcttcgc
tgggtctgcg acctgacgga gtcttccaac tcgccggatc 26940ggggagatct tccttcacgc
ctcgtcaggc cgtcctgact ttggagagtt cgtcctcgca 27000gccccgctcg ggcggcatcg
gcactctcca gttcgtggag gagttcactc cctcggtcta 27060cttcaacccc ttctccggct
cccccggcca ctacccggac gagttcatcc cgaacttcga 27120cgccatcagc gagtcggtgg
acggctacga ttgaatgtcc catggtggcg cggctgacct 27180agctcggctt cgacacctgg
accactgccg ccgcttccgc tgcttcgctc gggatctcgc 27240cgagtttgcc tactttgagc
tgcccgagga gcaccctcag ggcccggccc acggagtgcg 27300gatcgtcgtc gaagggggcc
tcgactccca cctgcttcgg attttcagcc agcgtccgat 27360cctggtcgag cgcgagcaag
gacagaccct tctgaccctg tactgcatct gcaaccaccc 27420cggcctgcat gaaagtcttt
gttgtctgct gtgtactgag tataataaaa gctgagatca 27480gcgactactc cggactcgat
tgtggtgttc ctgctatcaa ccggtccctg ttcttcaccg 27540ggaacgagac cgagctccag
ctccagtgta agccccacaa gaagtatctc acctggctgt 27600tccagggctc tccgatcgcc
gttgtcaacc actgcgacaa cgacggagtc ctgctgagcg 27660gccctgccaa ccttactttt
tccacccgca gaagcaagct ccagctcttc caacccttcc 27720tccccgggac ctatcagtgc
gtctcgggac cctgccatca caccttccac ctgatcccga 27780ataccacagc gccgctcccc
gctactaaca accaaactac ccaccaacgc caccgtcgcg 27840acctttctga atctaatact
accacccaca ccggaggtga gctccgaggt cgaccaacct 27900ctgggattta ctacggcccc
tgggaggtgg tagggttaat agcgctaggc ctagttgcgg 27960gtgggctttt ggctctctgc
tacctatacc tcccttgctg ttcgtactta gtggtgctgt 28020gttgctggtt taagaaatgg
ggaagatcac cctagtgagc tgcggtgtgc tggtggcggt 28080ggtgctttcg attgtgggac
tgggcggcgc ggctgtagtg aaggaggaga aggccgatcc 28140ctgcttgcat ttcaatcccg
acaaatgcca gctgagtttt cagcccgatg gcaatcggtg 28200cacggtgctg atcaagtgcg
gatgggaatg cgagaacgtg agaatcgagt acaataacaa 28260gactcggaac aatactctcg
cgtccgtgtg gcagcccggg gaccccgagt ggtacaccgt 28320ctctgtcccc ggtgctgacg
gctccccgcg caccgtgaac aatactttca tttttgcgca 28380catgtgcgac acggtcatgt
ggatgagcaa gcagtacgat atgtggcccc ccacgaagga 28440gaacatcgtg gtcttctcca
tcgcttacag cctgtgcacg gcgctaatca ccgctatcgt 28500gtgcctgagc attcacatgc
tcatcgctat tcgccccaga aataatgccg aaaaagagaa 28560acagccataa cacgtttttt
cacacacctt tttcagacca tggcctctgt tactgcccta 28620actatttttt tgggccttgt
gggtactagc agcacttttc agcatataaa caaaactgtt 28680tatgctggtt ctaattctgt
attacctggg catcaatcac accagaaagt ttcatggtac 28740tggtatgata aaagtaacac
gccagtcaca ctctgcaagg gtcatcaaac acccataaac 28800cgtagtggaa ttttttttaa
atgtaatcat aataatatta cactactttc aattacaaag 28860cactattctg gtacttacta
tggaaccaat tttaacataa aacaggacac ttactatagt 28920gtcacagtat tggatccaac
tactcctaga acaactacaa aacccacaac tactaagagg 28980cacactaaac ctaaaactac
caagaaaacc actgtcaaaa ctacaacaac taggaccacc 29040acaactacag aggctaccac
cagcacaaca cttgctgcca ctacacacac acacactgag 29100ctaaccttac agaccactaa
tgatttgatc gccctgttgc aaaaggggga taacagcacc 29160acttccaatg aggagatacc
cagatccatg attggcatta ttgttgctgt agtggtgtgc 29220atgttgatca tcgccttgtg
catggtgtac tatgccttct gctacagaaa gcacagactg 29280aacgacaagc tggaacactt
actaagtgtt gaattttaat tttttagaac catgaagatc 29340ctaggccttt ttagtttttc
tatcattacc tctactcttt gtgaatcagt ggataaagat 29400gttactatta ccactggttc
taattataca ctgaaagggc caccctcagg tatgctttcg 29460tggtattgct attttggaac
tgacactgat caaactgaat tatgcaattt tcaaaaaggc 29520aaaacctcaa actctaaaat
ctctaattat caatgcaatg gcactgatct gatactactc 29580aatgtcacga aagcatatgg
tggcagttat tcttgccctg gacaaaacac tgaggatatg 29640attttttaca aagtggaagt
ggttgatccc actactccac cgcccaccac cacaactact 29700cacaccacac acacagaaca
aacaccagag gcagcagaag cagagttggc cttccaggtt 29760cacggagatt cctttgctgt
caatacccct acacccgatc agcggtgtcc ggggctgctc 29820gtcagcggca ttgtcggtgt
gctttcggga ttagcagtca taatcatctg catgttcatt 29880tttgcttgct gctatagaag
gctttaccga caaaaatcag acccactgct gaacctctat 29940gtttaatttt ttccagagcc
atgaaggcag ttagcgctct agttttttgt tctttgattg 30000gcattgtttt tagtgctggg
tttttgaaaa atcttaccat ttatgaaggt gagaatgcca 30060ctctagtggg catcagtggt
caaaatgtca gctggctaaa ataccatcta gatgggtgga 30120aagacatttg cgattggaat
gtcactgtgt atacatgtaa tggagttaac ctcaccatta 30180ctaatgccac ccaagatcag
aatggtaggt ttaagggcca gagtttcact agaaataatg 30240ggtatgaatc ccataacatg
tttatctatg acgtcactgt catcagaaat gagactgcca 30300ccaccacaca gatgcccact
acacacagtt ctaccactac taccatgcaa accacacaga 30360caaccacttt ttatacatca
actcagcata tgaccaccac tacagcagca aagccaagta 30420gtgcagcgcc tcagccccag
gctttggctt tgatagctgc acaacctagt acaactacta 30480ggaccaatga gcagactact
gattttttgt ccactgtcga gagccacacc acagctacct 30540ccagtgcctt ctctagcacc
gccaatctct cctcgctttc ctctacacca atcagtcccg 30600ctactactac tcctagcccc
gctcctcttc ccactcccct gaagcaaact gaggacagcg 30660gcatgcaatg gcagatcacc
ctgctcattg tgatcgggtt ggtcattctg gccgtgttgc 30720tctactacat cttctgccgc
cgcattccca acgcgcaccg caagccggtc tacaagccca 30780tcgttgacgg gcaaccggag
ccgcttcagg tggaaggggg tctaaggaat cttctcttct 30840cttttacagt atggtgattg
aactatgatt cctagacaat tcttgatcac tattcttatc 30900tgcctcctcc aagtctgtgc
caccctcgct ctggtggcca acgccagtcc agactgtatt 30960gggcccttcg cctcctacgt
gctctttgcc ttcgtcacct gcatctgctg ctgtagcata 31020gtctgcctgc ttatcacctt
cttccagttc attgactgga tctttgtgcg catcgcctac 31080ctgcgccacc acccccagta
ccgcgaccag cgagtggcgc ggctgctcag gctcctctga 31140taagcatgcg ggctctgcta
cttctcgcgc ttctgctgtt agtgctcccc cgtcccgtca 31200acccccggtc ccccactcag
tcccccgagg aggtccgcaa atgcaaattc caagaaccct 31260ggaaattcct caaatgctac
cgccaaaaat cagacatgca tcccagctgg atcatgatca 31320ttgggatcgt gaacattctg
gcctgcaccc tcatctcctt tgtgatttac ccctgctttg 31380actttggttg gaactcgcca
gaggcgctct atctcccgcc tgaacctgac acaccaccac 31440agcaacctca ggcacacgca
ctaccaccac cacagcctag gccacaatac atgcccatat 31500tagactatga ggccgagcca
cagcgaccca tgctccccgc tattagttac ttcaatctaa 31560ccggcggaga tgactgaccc
actggccaac aacaacgtca acgaccttct cctggacatg 31620gacggccgcg cctcggagca
gcgactcgcc caacttcgca ttcgccagca gcaggagaga 31680gccgtcaagg agctgcagga
cggcatagcc atccaccagt gcaagaaagg catcttctgc 31740ctggtgaaac aggccaagat
ctcctacgag gtcacccaga ccgaccatcg cctctcctac 31800gagctcctgc agcagcgcca
gaagttcacc tgcctggtcg gagtcaaccc catcgtcatc 31860acccagcagt cgggcgatac
caaggggtgc atccactgct cctgcgactc ccccgactgc 31920gtccacactc tgatcaagac
cctctgcggc ctccgcgacc tcctccccat gaactaatca 31980cccccttatc cagtgaaata
aagatcatat tgatgatgat tttacagaaa taaagataca 32040atcatattga tgatttgagt
ttaataaaaa ataaagaatc acttacttga aatctgatac 32100caggtctctg tccatgtttt
ctgccaacac cacttcactc ccctcttccc agctctggta 32160ctgcaggccc cggcgggctg
caaacttcct ccacacgctg aaggggatgt caaattcctc 32220ctgtccctca atcttcattt
tatcttctat cagatgtcca aaaagcgcgt ccgggtggat 32280gatgacttcg accccgtcta
cccctacgat gcagacaacg caccgaccgt gcccttcatc 32340aaccccccct tcgtctcttc
agatggattc caagagaagc ccctgggggt gctgtccctg 32400cgactggccg accccgtcac
caccaagaac ggggaaatca ccctcaagct gggagagggg 32460gtggacctcg actcctcggg
aaaactcatc tccaacacgg ccaccaaggc cgccgcccct 32520ctcagttttt ccaacaacac
catttccctt aacatggatc acccctttta cactaaagat 32580ggaaaattat ccttacaagt
ttctccacca ttaaatatac tgagaacaag cattctaaac 32640acactagctt taggttttgg
atcaggttta ggactccgtg gctctgcctt ggcagtacag 32700ttagtctctc cacttacatt
tgatactgat ggaaacataa agcttacctt agacagaggt 32760ttgcatgtta caacaggaga
tgcaattgaa agcaacataa gctgggctaa aggtttaaaa 32820tttgaagatg gagccatagc
aaccaacatt ggaaatgggt tagagtttgg aagcagtagt 32880acagaaacag gtgttgatga
tgcttaccca atccaagtta aacttggatc tggccttagc 32940tttgacagta caggagccat
aatggctggt aacaaagaag acgataaact cactttgtgg 33000acaacacctg atccatcgcc
aaactgtcaa atactcgcag aaaatgatgc aaaactaaca 33060ctttgcttga ctaaatgtgg
tagtcaaata ctggccactg tgtcagtctt agttgtagga 33120agtggaaacc taaaccccat
tactggcacc gtaagcagtg ctcaggtgtt tctacgtttt 33180gatgcaaacg gtgttctttt
aacagaacat tctacactaa aaaaatactg ggggtatagg 33240cagggagata gcatagatgg
cactccatat accaatgctg taggattcat gcccaattta 33300aaagcttatc caaagtcaca
aagttctact actaaaaata atatagtagg gcaagtatac 33360atgaatggag atgtttcaaa
acctatgctt ctcactataa ccctcaatgg tactgatgac 33420agcaacagta catattcaat
gtcattttca tacacctgga ctaatggaag ctatgttgga 33480gcaacatttg gggctaactc
ttataccttc tcatacatcg cccaagaatg aacactgtat 33540cccaccctgc atgccaaccc
ttcccacccc actctgtgga aaaaactctg aaacacaaaa 33600taaaataaag ttcaagtgtt
ttattgattc aacagtttta caggattcga gcagttattt 33660ttcctccacc ctcccaggac
atggaataca ccaccctctc cccccgcaca gccttgaaca 33720tctgaacgcc attggtgatg
gacatgcttt tggtctccac gttccacaca gtttcagagc 33780gagccagtct cgggtcggtc
agggagatga aaccctccgg gcactcccgc atctgcacct 33840cacagctcaa cagctgagga
ttgtcctcgg tggtcgggat cacggttatc tggaagaagc 33900agaagagcgg cggtgggaat
catagtccgc gaacgggatc ggccggtggt gtcgcatcag 33960gccccgcagc agtcgctgtc
gccgccgctc cgtcaagctg ctgctcaggg ggtccgggtc 34020cagggactcc ctcagcatga
tgcccacggc cctcagcatc agtcgtctgg tgcggcgggc 34080gcagcagcgc atgcggatct
cgctcaggtc gctgcagtac gtgcaacaca ggaccaccag 34140gttgttcaac agtccatagt
tcaacacgct ccagccgaaa ctcatcgcgg gaaggatgct 34200acccacgtgg ccgtcgtacc
agatcctcag gtaaatcaag tggcgccccc tccagaacac 34260gctgcccatg tacatgatct
ccttgggcat gtggcggttc accacctccc ggtaccacat 34320caccctctgg ttgaacatgc
agccccggat gatcctgcgg aaccacaggg ccagcaccgc 34380cccgcccgcc atgcagcgaa
gagaccccgg gtcccggcaa tggcaatgga ggacccaccg 34440ctcgtacccg tggatcatct
gggagctgaa caagtctatg ttggcacagc acaggcacac 34500gctcatgcat ctcttcagca
ctctcagctc ctcgggggtc aaaaccatat cccagggcac 34560ggggaactct tgcaggacag
cgaaccccgc agaacagggc aatcctcgca cagaacttac 34620attgtgcatg gacagggtat
cgcaatcagg cagcaccggg tgatcctcca ccagggaagc 34680gcgggtctcg gtctcctcac
agcgtggtaa gggggccggc cgatacgggt gatggcggga 34740cgcggctgat cgtgctcgcg
accgtgtcat gatgcagttg ctttcggaca ttttcgtact 34800tgctgtagca gaacctggtc
cgggcgctgc acaccgatcg ccggcggcgg tcccggcgct 34860tggaacgctc ggtgttgaag
ttgtaaaaca gccactctct cagaccgtgc agcagatcta 34920gggcctcagg agtgatgaaa
atcccatcat gcctgatagc tctgatcaca tcgaccaccg 34980tggaatgggc cagacccagc
cagatgatgc aattttgttg ggtttcggta acggcggggg 35040agggaagaac aggaagaacc
atgattaact tttaatccaa acggtctcgg agcacttcaa 35100aatgaagatc gcggagatgg
cacctctcgc ccccgctgtg ttggtggaaa ataacagcca 35160ggtcaaaggt gatacggttc
tcgagatgtt ccacggtggc ttccagcaaa gcctccacgc 35220gcacatccag aaacaagaca
atagcgaaag cgggagggtt ctctaattcc tcaatcatca 35280tgttacactc ctgcaccatc
cccagataat tttcattttt ccagccttga atgattcgaa 35340ctagttcctg aggtaaatcc
aagccagcca tgataaagag ctcgcgcaga gcgccctcca 35400ccggcattct taagcacacc
ctcataattc caagatattc tgctcctggt tcacctgcag 35460cagattgaca agcggaatat
caaactctct gccgcgatcc ctaagctcct ccctcagcaa 35520taactgtaag tactctttca
tatcctctcc gaaattttta gccataggac cgccaggaat 35580aagattaggg caagccacag
tacagataaa ccgaagtcct ccccagtgag cattgccaaa 35640tgcaagactg ctataagcat
gctggctaga cccggtgata tcttccagat aactggacag 35700aaaatcgccc aggcaatttt
taagaaaatc aacaaaagaa aaatcctcca ggtgcacgtt 35760tagagcctcg ggaacaacga
tggagtaaat gcaagcggtg cgttccagca tggttagtta 35820gctgatctgt agaaaaaaca
aaaatgaaca ttaaaccatg ctagcctggc gaacaggtgg 35880gtaaatcgtt ctttccagca
ccaggcaggc cacggggtct ccggcacgac cctcgtaaaa 35940attgtcgcta tgattgaaaa
ccatcacaga gagacgttcc cggtggccgg cgtgaatgat 36000tcgacaagat gaatacaccc
ccggaacatt ggcgtccgcg agtgaaaaaa agcgcccgag 36060gaagcaataa ggcactacaa
tgctcagtct caagtccagc aaagcgatgc catgcggatg 36120aagcacaaaa ttctcaggtg
cgtacaaaat gtaattactc ccctcctgca caggcagcaa 36180agcccccgat ccctccaggt
acacatacaa agcctcagcg tccatagctt accgagcagc 36240agcagcagca cacaacaggc
gcaagagtca gagaaaggct gagctctaac ctgtccaccc 36300gctctctgct caatatatag
cccagatcta cactgacgta aaggccaaag tctaaaaata 36360cccgccaaat agtcacacac
gcccagcaca cgcccagaaa ccggtgacac actcaaaaaa 36420atacgcgcac ttcctcaaac
gcccaaactg ccgtcatttc cgggttccca cgctacgtca 36480tcaaaacacg actttcaaat
tccgtcgacc gttaaaaacg tcacccgccc cgcccctaac 36540ggtcgcccgt ctctcagcca
atcagcgccc cgcatcccca aattcaaaca cctcatttgc 36600atattaacgc gcaccaaaag
tttgaggtat attattgatg atg 3664315540PRTPan troglodytes
15Met Met Arg Arg Val Tyr Pro Glu Gly Pro Pro Pro Ser Tyr Glu Ser1
5 10 15Val Met Gln Gln Ala Val
Ala Ala Ala Met Gln Pro Pro Leu Glu Ala 20 25
30Pro Tyr Val Pro Pro Arg Tyr Leu Ala Pro Thr Glu Gly
Arg Asn Ser 35 40 45Ile Arg Tyr
Ser Glu Leu Ala Pro Leu Tyr Asp Thr Thr Arg Leu Tyr 50
55 60Leu Val Asp Asn Lys Ser Ala Asp Ile Ala Ser Leu
Asn Tyr Gln Asn65 70 75
80Asp His Ser Asn Phe Leu Thr Thr Val Val Gln Asn Asn Asp Phe Thr
85 90 95Pro Thr Glu Ala Ser Thr
Gln Thr Ile Asn Phe Asp Glu Arg Ser Arg 100
105 110Trp Gly Gly Gln Leu Lys Thr Ile Met His Thr Asn
Met Pro Asn Val 115 120 125Asn Glu
Phe Met Tyr Ser Asn Lys Phe Lys Ala Arg Val Met Val Ser 130
135 140Arg Lys Thr Pro Asn Gly Val Thr Val Gly Asp
Asp Tyr Asp Gly Ser145 150 155
160Gln Asp Glu Leu Thr Tyr Glu Trp Val Glu Phe Glu Leu Pro Glu Gly
165 170 175Asn Phe Ser Val
Thr Met Thr Ile Asp Leu Met Asn Asn Ala Ile Ile 180
185 190Asp Asn Tyr Leu Ala Val Gly Arg Gln Asn Gly
Val Leu Glu Ser Asp 195 200 205Ile
Gly Val Lys Phe Asp Thr Arg Asn Phe Arg Leu Gly Trp Asp Pro 210
215 220Val Thr Glu Leu Val Met Pro Gly Val Tyr
Thr Asn Glu Ala Phe His225 230 235
240Pro Asp Ile Val Leu Leu Pro Gly Cys Gly Val Asp Phe Thr Glu
Ser 245 250 255Arg Leu Ser
Asn Leu Leu Gly Ile Arg Lys Arg Gln Pro Phe Gln Glu 260
265 270Gly Phe Gln Ile Leu Tyr Glu Asp Leu Glu
Gly Gly Asn Ile Pro Ala 275 280
285Leu Leu Asp Val Glu Ala Tyr Glu Glu Ser Lys Glu Lys Ala Glu Ala 290
295 300Glu Ala Thr Thr Ala Val Ala Thr
Ala Ala Thr Val Ala Asp Ala Thr305 310
315 320Val Thr Arg Gly Asp Thr Phe Ala Thr Gln Ala Glu
Glu Ala Ala Ala 325 330
335Leu Ala Ala Thr Asp Asp Ser Glu Ser Lys Ile Val Ile Lys Pro Val
340 345 350Glu Lys Asp Ser Lys Asn
Arg Ser Tyr Asn Val Leu Pro Asp Gly Lys 355 360
365Asn Thr Ala Tyr Arg Ser Trp Tyr Leu Ala Tyr Asn Tyr Gly
Asp Pro 370 375 380Glu Lys Gly Val Arg
Ser Trp Thr Leu Leu Thr Thr Ser Asp Val Thr385 390
395 400Cys Gly Val Glu Gln Val Tyr Trp Ser Leu
Pro Asp Met Met Gln Asp 405 410
415Pro Val Thr Phe Arg Ser Thr Arg Gln Val Ser Asn Tyr Pro Val Val
420 425 430Gly Ala Glu Leu Leu
Pro Val Tyr Ser Lys Ser Phe Phe Asn Glu Gln 435
440 445Ala Val Tyr Ser Gln Gln Leu Arg Ala Phe Thr Ser
Leu Thr His Val 450 455 460Phe Asn Arg
Phe Pro Glu Asn Gln Ile Leu Val Arg Pro Pro Ala Pro465
470 475 480Thr Ile Thr Thr Val Ser Glu
Asn Val Pro Ala Leu Thr Asp His Gly 485
490 495Thr Leu Pro Leu Arg Ser Ser Ile Arg Gly Val Gln
Arg Val Thr Val 500 505 510Thr
Asp Ala Arg Arg Arg Thr Cys Pro Tyr Val Tyr Lys Ala Leu Gly 515
520 525Val Val Ala Pro Arg Val Leu Ser Ser
Arg Thr Phe 530 535 54016958PRTPan
troglodytes 16Met Tyr Val Arg Arg Pro Glu Gly Gly Arg Gly Ala Ser Pro Ser
Cys1 5 10 15Lys Met Ala
Thr Pro Ser Met Leu Pro Gln Trp Ala Tyr Met His Ile 20
25 30Ala Gly Gln Asp Ala Ser Glu Tyr Leu Ser
Pro Gly Leu Val Gln Phe 35 40
45Ala Arg Ala Thr Asp Thr Tyr Phe Ser Leu Gly Asn Lys Phe Arg Asn 50
55 60Pro Thr Val Ala Pro Thr His Asp Val
Thr Thr Asp Arg Ser Gln Arg65 70 75
80Leu Thr Leu Arg Phe Val Pro Val Asp Arg Glu Asp Asn Thr
Tyr Ser 85 90 95Tyr Lys
Val Arg Tyr Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp 100
105 110Met Ala Ser Thr Tyr Phe Asp Ile Arg
Gly Val Leu Asp Arg Gly Pro 115 120
125Ser Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ser Leu Ala Pro Lys
130 135 140Gly Ala Pro Asn Thr Ser Gln
Trp Lys Asp Ser Asp Ser Lys Met His145 150
155 160Thr Phe Gly Val Ala Ala Met Pro Gly Val Val Gly
Lys Lys Ile Glu 165 170
175Ala Asp Gly Leu Pro Ile Gly Ile Asp Ser Ser Ser Gly Thr Asp Thr
180 185 190Ile Ile Tyr Ala Asp Lys
Thr Phe Gln Pro Glu Pro Gln Val Gly Ser 195 200
205Asp Ser Trp Val Asp Thr Asn Gly Ala Glu Glu Lys Tyr Gly
Gly Arg 210 215 220Ala Leu Lys Asp Thr
Thr Asn Met Lys Pro Cys Tyr Gly Ser Phe Ala225 230
235 240Arg Pro Thr Asn Lys Glu Gly Gly Gln Ala
Asn Ile Lys Asp Ser Glu 245 250
255Thr Ala Ser Thr Thr Pro Asn Tyr Asp Ile Asp Leu Ala Phe Phe Asp
260 265 270Ser Lys Asn Ile Ala
Ala Asn Tyr Asp Pro Asp Ile Val Met Tyr Thr 275
280 285Glu Asn Val Glu Leu Gln Thr Pro Asp Thr His Ile
Val Phe Lys Pro 290 295 300Gly Thr Ser
Asp Glu Ser Ser Glu Ala Asn Leu Gly Gln Gln Ala Met305
310 315 320Pro Asn Arg Pro Asn Tyr Ile
Gly Phe Arg Asp Asn Phe Ile Gly Leu 325
330 335Met Tyr Tyr Asn Ser Thr Gly Asn Met Gly Val Leu
Ala Gly Gln Ala 340 345 350Ser
Gln Leu Asn Ala Val Val Asp Leu Gln Asp Arg Asn Thr Glu Leu 355
360 365Ser Tyr Gln Leu Leu Leu Asp Ser Leu
Gly Asp Arg Thr Arg Tyr Phe 370 375
380Ser Met Trp Asn Gln Ala Val Asp Ser Tyr Asp Pro Asp Val Arg Ile385
390 395 400Ile Glu Asn His
Gly Val Glu Asp Glu Leu Pro Asn Tyr Cys Phe Pro 405
410 415Leu Asn Gly Val Gly Phe Thr Asp Thr Tyr
Gln Gly Val Lys Val Lys 420 425
430Thr Asp Thr Ala Ala Thr Gly Thr Asn Gly Thr Gln Trp Asp Lys Asp
435 440 445Asp Thr Thr Val Ser Thr Ala
Asn Glu Ile His Ser Gly Asn Pro Phe 450 455
460Ala Met Glu Ile Asn Ile Gln Ala Asn Leu Trp Arg Asn Phe Leu
Tyr465 470 475 480Ala Asn
Val Ala Leu Tyr Leu Pro Asp Ser Tyr Lys Tyr Thr Pro Ala
485 490 495Asn Ile Thr Leu Pro Thr Asn
Thr Asn Thr Tyr Asp Tyr Met Asn Gly 500 505
510Arg Val Val Ala Pro Ser Leu Val Asp Ala Tyr Ile Asn Ile
Gly Ala 515 520 525Arg Trp Ser Leu
Asp Pro Met Asp Asn Val Asn Pro Phe Asn His His 530
535 540Arg Asn Ala Gly Leu Arg Tyr Arg Ser Met Leu Leu
Gly Asn Gly Arg545 550 555
560Tyr Val Pro Phe His Ile Gln Val Pro Gln Lys Phe Phe Ala Ile Lys
565 570 575Ser Leu Leu Leu Leu
Pro Gly Ser Tyr Thr Tyr Glu Trp Asn Phe Arg 580
585 590Lys Asp Val Asn Met Ile Leu Gln Ser Ser Leu Gly
Asn Asp Leu Arg 595 600 605Thr Asp
Gly Ala Ser Ile Ala Phe Thr Ser Ile Asn Leu Tyr Ala Thr 610
615 620Phe Phe Pro Met Ala His Asn Thr Ala Ser Thr
Leu Glu Ala Met Leu625 630 635
640Arg Asn Asp Thr Asn Asp Gln Ser Phe Asn Asp Tyr Leu Ser Ala Ala
645 650 655Asn Met Leu Tyr
Pro Ile Pro Ala Asn Ala Thr Asn Val Pro Ile Ser 660
665 670Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg Gly
Trp Ser Phe Thr Arg 675 680 685Leu
Lys Thr Arg Glu Thr Pro Ser Leu Gly Ser Gly Phe Asp Pro Tyr 690
695 700Phe Val Tyr Ser Gly Ser Ile Pro Tyr Leu
Asp Gly Thr Phe Tyr Leu705 710 715
720Asn His Thr Phe Lys Lys Val Ser Ile Thr Phe Asp Ser Ser Val
Ser 725 730 735Trp Pro Gly
Asn Asp Arg Leu Leu Thr Pro Asn Glu Phe Glu Ile Lys 740
745 750Arg Thr Val Asp Gly Glu Gly Tyr Asn Val
Ala Gln Cys Asn Met Thr 755 760
765Lys Asp Trp Phe Leu Val Gln Met Leu Ala His Tyr Asn Ile Gly Tyr 770
775 780Gln Gly Phe Tyr Val Pro Glu Gly
Tyr Lys Asp Arg Met Tyr Ser Phe785 790
795 800Phe Arg Asn Phe Gln Pro Met Ser Arg Gln Val Val
Asp Glu Val Asn 805 810
815Tyr Lys Asp Tyr Gln Ala Val Thr Leu Ala Tyr Gln His Asn Asn Ser
820 825 830Gly Phe Val Gly Tyr Leu
Ala Pro Thr Met Arg Gln Gly Gln Pro Tyr 835 840
845Pro Ala Asn Tyr Pro Tyr Pro Leu Ile Gly Lys Ser Ala Val
Ala Ser 850 855 860Val Thr Gln Lys Lys
Phe Leu Cys Asp Arg Val Met Trp Arg Ile Pro865 870
875 880Phe Ser Ser Asn Phe Met Ser Met Gly Ala
Leu Thr Asp Leu Gly Gln 885 890
895Asn Met Leu Tyr Ala Asn Ser Ala His Ala Leu Asp Met Asn Phe Glu
900 905 910Val Asp Pro Met Asp
Glu Ser Thr Leu Leu Tyr Val Val Phe Glu Val 915
920 925Phe Asp Val Val Arg Val His Gln Pro His Arg Gly
Val Ile Glu Ala 930 935 940Val Tyr Leu
Arg Thr Pro Phe Ser Ala Gly Asn Ala Thr Thr945 950
95517425PRTPan troglodytes 17Met Ser Lys Lys Arg Val Arg Val Asp
Asp Asp Phe Asp Pro Val Tyr1 5 10
15Pro Tyr Asp Ala Asp Asn Ala Pro Thr Val Pro Phe Ile Asn Pro
Pro 20 25 30Phe Val Ser Ser
Asp Gly Phe Gln Glu Lys Pro Leu Gly Val Leu Ser 35
40 45Leu Arg Leu Ala Asp Pro Val Thr Thr Lys Asn Gly
Glu Ile Thr Leu 50 55 60Lys Leu Gly
Glu Gly Val Asp Leu Asp Ser Ser Gly Lys Leu Ile Ser65 70
75 80Asn Thr Ala Thr Lys Ala Ala Ala
Pro Leu Ser Phe Ser Asn Asn Thr 85 90
95Ile Ser Leu Asn Met Asp His Pro Phe Tyr Thr Lys Asp Gly
Lys Leu 100 105 110Ser Leu Gln
Val Ser Pro Pro Leu Asn Ile Leu Arg Thr Ser Ile Leu 115
120 125Asn Thr Leu Ala Leu Gly Phe Gly Ser Gly Leu
Gly Leu Arg Gly Ser 130 135 140Ala Leu
Ala Val Gln Leu Val Ser Pro Leu Thr Phe Asp Thr Asp Gly145
150 155 160Asn Ile Lys Leu Thr Leu Asp
Arg Gly Leu His Val Thr Thr Gly Asp 165
170 175Ala Ile Glu Ser Asn Ile Ser Trp Ala Lys Gly Leu
Lys Phe Glu Asp 180 185 190Gly
Ala Ile Ala Thr Asn Ile Gly Asn Gly Leu Glu Phe Gly Ser Ser 195
200 205Ser Thr Glu Thr Gly Val Asp Asp Ala
Tyr Pro Ile Gln Val Lys Leu 210 215
220Gly Ser Gly Leu Ser Phe Asp Ser Thr Gly Ala Ile Met Ala Gly Asn225
230 235 240Lys Glu Asp Asp
Lys Leu Thr Leu Trp Thr Thr Pro Asp Pro Ser Pro 245
250 255Asn Cys Gln Ile Leu Ala Glu Asn Asp Ala
Lys Leu Thr Leu Cys Leu 260 265
270Thr Lys Cys Gly Ser Gln Ile Leu Ala Thr Val Ser Val Leu Val Val
275 280 285Gly Ser Gly Asn Leu Asn Pro
Ile Thr Gly Thr Val Ser Ser Ala Gln 290 295
300Val Phe Leu Arg Phe Asp Ala Asn Gly Val Leu Leu Thr Glu His
Ser305 310 315 320Thr Leu
Lys Lys Tyr Trp Gly Tyr Arg Gln Gly Asp Ser Ile Asp Gly
325 330 335Thr Pro Tyr Thr Asn Ala Val
Gly Phe Met Pro Asn Leu Lys Ala Tyr 340 345
350Pro Lys Ser Gln Ser Ser Thr Thr Lys Asn Asn Ile Val Gly
Gln Val 355 360 365Tyr Met Asn Gly
Asp Val Ser Lys Pro Met Leu Leu Thr Ile Thr Leu 370
375 380Asn Gly Thr Asp Asp Ser Asn Ser Thr Tyr Ser Met
Ser Phe Ser Tyr385 390 395
400Thr Trp Thr Asn Gly Ser Tyr Val Gly Ala Thr Phe Gly Ala Asn Ser
405 410 415Tyr Thr Phe Ser Tyr
Ile Ala Gln Glu 420 4251830971DNAArtificial
SequenceM72-ChAd63 construct 18catcatcaat aatatacctc aaacttttgg
tgcgcgttaa tatgcaaatg aggtgtttga 60atttggggat gcggggcgct gattggctga
gagacgggcg accgttaggg gcggggcggg 120tgacgttttg atgacgtggc cgtgaggcgg
agccggtttg caagttctcg tgggaaaagt 180gacgtcaaac gaggtgtggt ttgaacacgg
aaatactcaa ttttcccgcg ctctctgaca 240ggaaatgagg tgtttctggg cggatgcaag
tgaaaacggg ccattttcgc gcgaaaactg 300aatgaggaag tgaaaatctg agtaattccg
cgtttatggc agggaggagt atttgccgag 360ggccgagtag actttgaccg attacgtggg
ggtttcgatt accgtatttt tcacctaaat 420ttccgcgtac ggtgtcaaag tccggtgttt
ttacggatat cccattgcat acgttgtatc 480catatcataa tatgtacatt tatattggct
catgtccaac attaccgcca tgttgacatt 540gattattgac tagttattaa tagtaatcaa
ttacggggtc attagttcat agcccatata 600tggagttccg cgttacataa cttacggtaa
atggcccgcc tggctgaccg cccaacgacc 660cccgcccatt gacgtcaata atgacgtatg
ttcccatagt aacgccaata gggactttcc 720attgacgtca atgggtggag tatttacggt
aaactgccca cttggcagta catcaagtgt 780atcatatgcc aagtacgccc cctattgacg
tcaatgacgg taaatggccc gcctggcatt 840atgcccagta catgacctta tgggactttc
ctacttggca gtacatctac gtattagtca 900tcgctattac catggtgatg cggttttggc
agtacatcaa tgggcgtgga tagcggtttg 960actcacgggg atttccaagt ctccacccca
ttgacgtcaa tgggagtttg ttttggcacc 1020aaaatcaacg ggactttcca aaatgtcgta
acaactccgc cccattgacg caaatgggcg 1080gtaggcgtgt acggtgggag gtctatataa
gcagagctct ccctatcagt gatagagatc 1140tccctatcag tgatagagat cgtcgacgag
ctcgtttagt gaaccgtcag atcgcctgga 1200gacgccatcc acgctgtttt gacctccata
gaagacaccg ggaccgatcc agcctccgcg 1260gccgggaacg gtgcattgga acgcggattc
cccgtgccaa gagtgagatc ttccgtttat 1320ctaggtacca gatatcgcca ccatgaccgc
cgccagcgac aacttccagc tgtctcaggg 1380cggccagggc ttcgccatcc ctatcggcca
agctatggcc attgctggac agatcagaag 1440cggcggaggc agccctaccg tgcatatcgg
ccctaccgcc ttcctgggcc tgggcgtggt 1500ggacaacaac ggcaacggcg ccagagtgca
gcgggtggtc ggatctgccc ctgccgcaag 1560cctgggcatc agcaccgggg atgtgatcac
cgccgtggat ggcgccccta tcaacagcgc 1620cacagccatg gccgacgccc tgaatggaca
ccaccccggc gacgtgatca gcgtgacctg 1680gcagaccaag agcggaggca ccagaaccgg
caacgtgaca ctggccgagg gacctcccgc 1740cgagttcatg gtggatttcg gcgccctgcc
ccccgagatc aactccgcca ggatgtatgc 1800cggccctggc agcgcctctc tggtggccgc
tgctcagatg tgggacagcg tggccagcga 1860tctgttcagc gccgcctccg ccttccagtc
cgtggtctgg ggcctgaccg tgggcagctg 1920gatcggaagc agtgccggcc tgatggtggc
tgccgcctct ccctacgtgg cctggatgtc 1980agtcacagcc ggccaggccg aactgactgc
cgctcaagtg cgagtggctg ctgctgccta 2040tgagacagcc tacggcctga cagtgccccc
acccgtgatc gccgagaacc gggccgagct 2100gatgatcctg atcgccacca acctgctggg
ccagaacacc cccgccattg ccgtgaacga 2160ggccgagtac ggcgagatgt gggcccagga
cgccgctgcc atgtttggct atgccgctgc 2220tacagccacc gccactgcca ccctgctgcc
cttcgaagag gcccccgaga tgacctctgc 2280cggcggactg ctggaacagg ccgctgccgt
ggaagaggcc agcgacacag ccgccgctaa 2340ccagctgatg aacaacgtgc cccaggccct
gcagcagctg gcacagccta cacagggcac 2400caccccttct agcaagctcg gcggcctgtg
gaaaaccgtg tccccccacc ggtcccccat 2460cagcaacatg gtgtccatgg ccaacaacca
catgagcatg accaacagcg gcgtgtccat 2520gaccaatacc ctgagcagca tgctgaaggg
ctttgcccca gccgctgccg ctcaggctgt 2580gcagacagct gctcagaatg gcgtgcgggc
catgagcagc ctgggcagtt ccctgggcag 2640ctctggactg ggagggggcg tggccgccaa
tctgggcaga gccgctagcg tgggcagcct 2700gtctgtgcct caagcctggg ctgctgccaa
tcaggccgtg acaccagccg ctagagccct 2760gcctctgacc agcctgacct ctgctgccga
gaggggccct ggccagatgc tgggaggact 2820gcctgtgggc cagatgggag ccagagccgg
cggaggactg agcggcgtgc tgagagtgcc 2880ccccagaccc tacgtgatgc cccactctcc
cgccgctggc gatattgccc ctcccgccct 2940gagccaggac agattcgccg acttccctgc
cctgcccctg gatccttctg ccatggtggc 3000tcaagtggga ccccaggtgg tgaacatcaa
caccaagctg ggctacaaca acgccgtggg 3060agccggcacc ggcatcgtga tcgaccccaa
tggcgtggtg ctgaccaaca atcacgtgat 3120cgctggcgcc accgacatca acgccttcag
cgtgggctcc ggccagacct acggcgtgga 3180cgtggtcgga tacgaccgga cccaggatgt
ggccgtgctg cagctgagag gcgctggcgg 3240actgccttct gccgccattg gaggcggagt
ggccgtggga gaacctgtgg tggccatggg 3300caatagcggc ggacagggcg gcacacctag
agctgtgcct ggaagagtgg tggccctggg 3360acagaccgtg caggccagcg atagcctgac
aggcgccgag gaaaccctga acggcctgat 3420ccagttcgac gccgccatcc agcctgggga
tgctggcgga cctgtggtga acggactggg 3480ccaggtggtc ggaatgaata ccgccgcctc
ctaatagtga gcggccgcga tctgctgtgc 3540cttctagttg ccagccatct gttgtttgcc
cctcccccgt gccttccttg accctggaag 3600gtgccactcc cactgtcctt tcctaataaa
atgaggaaat tgcatcgcat tgtctgagta 3660ggtgtcattc tattctgggg ggtggggtgg
ggcaggacag caagggggag gattgggaag 3720acaatagcag gcatgctggg gatgcggtgg
gctctagata tcagcgatcg cgtgagtagt 3780gtttgggggt gggtgggagc ctgcatgatg
ggcagaatga ctaaaatctg tgtttttctg 3840tgtgttgcag cagcatgagc ggaagcgcct
cctttgaggg aggggtattc agcccttatc 3900tgacggggcg tctcccctcc tgggcgggag
tgcgtcagaa tgtgatggga tccacggtgg 3960acggccggcc cgtgcagccc gcgaactctt
caaccctgac ctacgcgacc ctgagctcct 4020cgtccgtgga cgcagctgcc gccgcagctg
ctgcttccgc cgccagcgcc gtgcgcggaa 4080tggccctggg cgccggctac tacagctctc
tggtggccaa ctcgagttcc accaataatc 4140ccgccagcct gaacgaggag aagctgttgc
tgctgatggc ccagctcgag gccctgaccc 4200agcgcctggg cgagctgacc cagcaggtgg
ctcagctgca ggcggagacg cgggccgcgg 4260ttgccacggt gaaaaccaaa taaaaaatga
atcaataaat aaacggagac ggttgttgat 4320tttaacacag agtcttgaat ctttatttga
tttttcgcgc gcggtaggcc ctggaccacc 4380ggtctcgatc attgagcacc cggtggatct
tttccaggac ccggtagagg tgggcttgga 4440tgttgaggta catgggcatg agcccgtccc
gggggtggag gtagctccat tgcagggcct 4500cgtgctcggg ggtggtgttg taaatcaccc
agtcatagca ggggcgcagg gcgtggtgct 4560gcacgatgtc tttgaggagg agactgatgg
ccacgggcag ccccttggtg taggtgttga 4620cgaacctatt gagctgggag ggatgcatgc
ggggggagat gagatgcatc ttggcctgga 4680tcttgagatt ggcgatgttc ccgcccagat
cccgccgggg gttcatgttg tgcaggacca 4740ccagcacggt gtatccggtg cacttgggga
atttgtcatg caacttggaa gggaaggcgt 4800gaaagaattt ggagacgccc ttgtgaccgc
ccaggttttc catgcactca tccatgatga 4860tggcgatggg cccgtgggcg gcggcctggg
caaagacgtt tcgggggtcg gacacatcgt 4920agttgtggtc ctgggtgagc tcgtcatagg
ccattttaat gaatttgggg cggagggtac 4980ccgactgggg gacaaaggtg ccctcgatcc
cgggggcgta gttcccctcg cagatctgca 5040tctcccaggc cttgagctcg gaggggggga
tcatgtccac ctgcggggcg atgaaaaaaa 5100cggtttccgg ggcgggggag atgagctgcg
ccgaaagcag gttccggagc agctgggact 5160tgccgcagcc ggtggggccg tagatgaccc
cgatgaccgg ctgcaggtgg tagttgaggg 5220agagacagct gccgtcctcg cggaggaggg
gggccacctc gttcatcatc tcgcgcacat 5280gcatgttctc gcgcacgagt tccgccagga
ggcgctcgcc ccccagcgag aggagctctt 5340gcagcgaggc gaagtttttc agcggcttga
gcccgtcggc catgggcatt ttggagaggg 5400tctgttgcaa gagttccaga cggtcccaga
gctcggtgat gtgctctagg gcatctcgat 5460ccagcagacc tcctcgtttc gcgggttggg
gcgactgcgg gagtagggca ccaggcgatg 5520ggcgtccagc gaggccaggg tccggtcctt
ccagggtcgc agggtccgcg tcagcgtggt 5580ctccgtcacg gtgaaggggt gcgcgccggg
ctgggcgctt gcgagggtgc gcttcaggct 5640catccggctg gtcgagaacc gctcccggtc
ggcgccctgc gcgtcggcca ggtagcaatt 5700gagcatgagt tcgtagttga gcgcctcggc
cgcgtggccc ttggcgcgga gcttaccttt 5760ggaagtgtgt ccgcagacgg gacagaggag
ggacttgagg gcgtagagct tgggggcgag 5820gaagacggac tcgggggcgt aggcgtccgc
gccgcagctg gcgcagacgg tctcgcactc 5880cacgagccag gtgaggtcgg ggcggtcggg
gtcaaaaacg aggtttcctc cgtgcttttt 5940gatgcgtttc ttacctctgg tctccatgag
ctcgtgtccc cgctgggtga caaagaggct 6000gtccgtgtcc ccgtagaccg actttatggg
ccggtcctcg agcggggtgc cgcggtcctc 6060gtcgtagagg aaccccgccc actccgagac
gaaggcccgg gtccaggcca gcacgaagga 6120ggccacgtgg gaggggtagc ggtcgttgtc
caccagcggg tccaccttct ccagggtatg 6180caagcacatg tccccctcgt ccacatccag
gaaggtgatt ggcttgtaag tgtaggccac 6240gtgaccgggg gtcccggccg ggggggtata
aaagggggcg ggcccctgct cgtcctcact 6300gtcttccgga tcgctgtcca ggagcgccag
ctgttggggt aggtattccc tctcgaaggc 6360gggcatgacc tcggcactca ggttgtcagt
ttctagaaac gaggaggatt tgatattgac 6420ggtgccgttg gagacgcctt tcatgagccc
ctcgtccatc tggtcagaaa agacgatctt 6480tttgttgtcg agcttggtgg cgaaggagcc
gtagagggcg ttggagagca gcttggcgat 6540ggagcgcatg gtctggttct tttccttgtc
ggcgcgctcc ttggcggcga tgttgagctg 6600cacgtactcg cgcgccacgc acttccattc
ggggaagacg gtggtgagct cgtcgggcac 6660gattctgacc cgccagccgc ggttgtgcag
ggtgatgagg tccacgctgg tggccacctc 6720gccgcgcagg ggctcgttgg tccagcagag
gcgcccgccc ttgcgcgagc agaagggggg 6780cagcgggtcc agcatgagct cgtcgggggg
gtcggcgtcc acggtgaaga tgccgggcag 6840gagctcgggg tcgaagtagc tgatgcaggt
gcccagatcg tccagacttg cttgccagtc 6900gcgcacggcc agcgcgcgct cgtaggggct
gaggggcgtg ccccagggca tggggtgcgt 6960gagcgcggag gcgtacatgc cgcagatgtc
gtagacgtag aggggctcct ggaggacgcc 7020gatgtaggtg gggtagcagc gccccccgcg
gatgctggcg cgcacgtagt cgtacagctc 7080gtgcgagggc gcgaggagcc ccgtgccgag
attggagcgc tgcggctttt cggcgcggta 7140gacgatctgg cggaagatgg cgtgggagtt
ggaggagatg gtgggcctct ggaagatgtt 7200gaagtgggca tggggcagtc cgaccgagtc
cctgatgaag tgggcgtagg agtcctgcag 7260cttggcgacg agctcggcgg tgacgaggac
gtccagggcg cagtagtcga gggtctcttg 7320gatgatgtcg tacttgagct ggcccttctg
cttccacagc tcgcggttga gaaggaactc 7380ttcgcggtcc ttccagtact cttcgagggg
gaacccgtcc tgatcggcac ggtaagagcc 7440caccatgtag aactggttga cggccttgta
ggcgcagcag cccttctcca cggggagggc 7500gtaagcttgc gcggccttgc gcagggaggt
gtgggtgagg gcgaaggtgt cgcgcaccat 7560gactttgagg aactggtgct tgaagtcgag
gtcgtcgcag ccgccctgct cccagagctg 7620gaagtccgtg cgcttcttgt aggcggggtt
gggcaaagcg aaagtaacat cgttgaagag 7680gatcttgccc gcgcggggca tgaagttgcg
agtgatgcgg aaaggctggg gcacctcggc 7740ccggttgttg atgacctggg cggcgaggac
gatctcgtcg aagccgttga tgttgtgccc 7800gacgatgtag agttccacga atcgcgggcg
gcccttgacg tggggcagct tcttgagctc 7860gtcgtaggtg agctcggcgg ggtcgctgag
cccgtgctgc tcgagggccc agtcggcgac 7920gtgggggttg gcgctgagga aggaagtcca
gagatccacg gccagggcgg tctgcaagcg 7980gtcccggtac tgacggaact gctggcccac
ggccattttt tcgggggtga cgcagtagaa 8040ggtgcggggg tcgccgtgcc agcggtccca
cttgagctgg agggcgaggt cgtgggcgag 8100ctcgacgagc ggcgggtccc cggagagttt
catgaccagc atgaagggga cgagctgctt 8160gccgaaggac cccatccagg tgtaggtttc
cacatcgtag gtgaggaaga gcctttcggt 8220gcgaggatgc gagccgatgg ggaagaactg
gatctcctgc caccagttgg aggaatggct 8280gttgatgtga tggaagtaga aatgccgacg
gcgcgccgag cactcgtgct tgtgtttata 8340caagcgtccg cagtgctcgc aacgctgcac
gggatgcacg tgctgcacga gctgtacctg 8400ggttcctttg acgaggaatt tcagtgggca
gtggagcgct ggcggctgca tctggtgctg 8460tactacgtcc tggccatcgg cgtggccatc
gtctgcctcg atggtggtca tgctgacgag 8520cccgcgcggg aggcaggtcc agacctcggc
tcggacgggt cggagagcga ggacgagggc 8580gcgcaggccg gagctgtcca gggtcctgag
acgctgcgga gtcaggtcag tgggcagcgg 8640cggcgcgcgg ttgacttgca ggagcttttc
cagggcgcgc gggaggtcca gatggtactt 8700gatctccacg gcgccgttgg tggcgacgtc
cacggcttgc agggtcccgt gcccctgggg 8760cgccaccacc gtgccccgtt tcttcttggg
cggcggcggc tccatgctta gaagcggcgg 8820cgaggacgcg cgccgggcgg caggggcggc
tcggggcccg gaggcagggg cggcaggggc 8880acgtcggcgc cgcgcgcggg caggttctgg
tactgcgccc ggagaagact ggcgtgagcg 8940acgacgcgac ggttgacgtc ctggatctga
cgcctctggg tgaaggccac gggacccgtg 9000agtttgaacc tgaaagagag ttcgacagaa
tcaatctcgg tatcgttgac ggcggcctgc 9060cgcaggatct cttgcacgtc gcccgagttg
tcctggtagg cgatctcggt catgaactgc 9120tcgatctcct cctcctgaag gtctccgcgg
ccggcgcgct cgacggtggc cgcgaggtcg 9180ttggagatgc ggcccatgag ctgcgagaag
gcgttcatgc cggcctcgtt ccagacgcgg 9240ctgtagacca cggctccgtc ggggtcgcgc
gcgcgcatga ccacctgggc gaggttgagc 9300tcgacgtggc gcgtgaagac cgcgtagttg
cagaggcgct ggtagaggta gttgagcgtg 9360gtggcgatgt gctcggtgac gaagaagtac
atgatccagc ggcggagcgg catctcgctg 9420acgtcgccca gggcttccaa gcgctccatg
gcctcgtaga agtccacggc gaagttgaaa 9480aactgggagt tgcgcgccga gacggtcaac
tcctcctcca gaagacggat gagctcggcg 9540atggtggcgc gcacctcgcg ctcgaaggcc
ccggggggct cctcttccat ttcctcctct 9600tcctcctcca ctaacatctc ttctacttcc
tcctcaggag gcggcggcgg gggaggggcc 9660ctgcgtcgcc ggcggcgcac gggcagacgg
tcgatgaagc gctcgatggt ctccccgcgc 9720cggcgacgca tggtctcggt gacggcgcgc
ccgtcctcgc ggggccgcag cgtgaagacg 9780ccgccgcgca tctccaggtg gccgccgggg
gggtctccgt tgggcaggga gagggcgctg 9840acgatgcatc ttatcaattg acccgtaggg
actccgcgca aggacctgag cgtctcgaga 9900tccacgggat ccgaaaaccg ctgaacgaag
gcttcgagcc agtcgcagtc gcaaggtagg 9960ctgagcccgg tttcttgttc ttcgggtatt
tggtcgggag gcgggcgggc gatgctgctg 10020gtgatgaagt tgaagtaggc ggtcctgaga
cggcggatgg tggcgaggag caccaggtcc 10080ttgggcccgg cttgctggat gcgcagacgg
tcggccatgc cccaggcgtg gtcctgacac 10140ctggcgaggt ccttgtagta gtcctgcatg
agccgctcca cgggcacctc ctcctcgccc 10200gcgcggccgt gcatgcgcgt gagcccgaac
ccgcgctgcg gctggacgag cgccaggtcg 10260gcgacgacgc gctcggcgag gatggcctgc
tggatctggg tgagggtggt ctggaagtcg 10320tcgaagtcga cgaagcggtg gtaggctccg
gtgttgatgg tgtaggagca gttggccatg 10380acggaccagt tgacggtctg gtggccgggg
cgcacgagct cgtggtactt gaggcgcgag 10440taggcgcgcg tgtcgaagat gtagtcgttg
caggtgcgca cgaggtactg gtatccgacg 10500aggaagtgcg gcggcggctg gcggtagagc
ggccatcgct cggtggcggg ggcgccgggc 10560gcgaggtcct cgagcatgag gcggtggtag
ccgtagatgt acctggacat ccaggtgatg 10620ccggcggcgg tggtggaggc gcgcgggaac
tcgcggacgc ggttccagat gttgcgcagc 10680ggcaggaagt agttcatggt ggccgcggtc
tggcccgtga ggcgcgcgca gtcgtggatg 10740ctctagacat acgggcaaaa acgaaagcgg
tcagcggctc gactccgtgg cctggaggct 10800aagcgaacgg gttgggctgc gcgtgtaccc
cggttcgaat ctcgaatcag gctggagccg 10860cagctaacgt ggtactggca ctcccgtctc
gacccaagcc tgctaacgaa acctccagga 10920tacggaggcg ggtcgttttt tggccttggt
cgctggtcat gaaaaactag taagcgcgga 10980aagcggccgc ccgcgatggc tcgctgccgt
agtctggaga aagaatcgcc agggttgcgt 11040tgcggtgtgc cccggttcga gcctcagcgc
tcggcgccgg ccggattccg cggctaacgt 11100gggcgtggct gccccgtcgt ttccaagacc
ccttagccag ccgacttctc cagttacgga 11160gcgagcccct ctttttttct tgtgtttttg
ccagatgcat cccgtactgc ggcagatgcg 11220cccccaccct ccaccacaac cgcccctacc
gcagcagcag caacagccgg cgcttctgcc 11280cccgccccag cagcagcagc cagccactac
cgcggcggcc gccgtgagcg gagccggcgt 11340tcagtatgac ctggccttgg aagagggcga
ggggctggcg cggctggggg cgtcgtcgcc 11400ggagcggcac ccgcgcgtgc agatgaaaag
ggacgctcgc gaggcctacg tgcccaagca 11460gaacctgttc agagacagga gcggcgagga
gcccgaggag atgcgcgcct cccgcttcca 11520cgcggggcgg gagctgcggc gcggcctgga
ccgaaagcgg gtgctgaggg acgaggattt 11580cgaggcggac gagctgacgg ggatcagccc
cgcgcgcgcg cacgtggccg cggccaacct 11640ggtcacggcg tacgagcaga ccgtgaagga
ggagagcaac ttccaaaaat ccttcaacaa 11700ccacgtgcgc acgctgatcg cgcgcgagga
ggtgaccctg ggcctgatgc acctgtggga 11760cctgctggag gccatcgtgc agaaccccac
gagcaagccg ctgacggcgc agctgtttct 11820ggtggtgcag cacagtcggg acaacgagac
gttcagggag gcgctgctga atatcaccga 11880gcccgagggc cgctggctcc tggacctggt
gaacattctg cagagcatcg tggtgcagga 11940gcgcgggctg ccgctgtccg agaagctggc
ggccatcaac ttctcggtgc tgagcctggg 12000caagtactac gctaggaaga tctacaagac
cccgtacgtg cccatagaca aggaggtgaa 12060gatcgatggg ttttacatgc gcatgaccct
gaaagtgctg accctgagcg acgatctggg 12120ggtgtaccgc aacgacagga tgcaccgcgc
ggtgagcgcc agccgccggc gcgagctgag 12180cgaccaggag ctgatgcaca gcctgcagcg
ggccctgacc ggggccggga ccgaggggga 12240gagctacttt gacatgggcg cggacctgcg
ctggcagccc agccgccggg ccttggaagc 12300tgccggcggc gtgccctacg tggaggaggt
ggacgatgag gaggaggagg gcgagtacct 12360ggaagactga tggcgcgacc gtatttttgc
tagatgcagc aacagccacc gccgccgcct 12420cctgatcccg cgatgcgggc ggcgctgcag
agccagccgt ccggcattaa ctcctcggac 12480gattggaccc aggccatgca acgcatcatg
gcgctgacga cccgcaatcc cgaagccttt 12540agacagcagc ctcaggccaa ccggctctcg
gccatcctgg aggccgtggt gccctcgcgc 12600tcgaacccca cgcacgagaa ggtgctggcc
atcgtgaacg cgctggtgga gaacaaggcc 12660atccgcggcg acgaggccgg gctggtgtac
aacgcgctgc tggagcgcgt ggcccgctac 12720aacagcacca acgtgcagac gaacctggac
cgcatggtga ccgacgtgcg cgaggcggtg 12780tcgcagcgcg agcggttcca ccgcgagtcg
aacctgggct ccatggtggc gctgaacgcc 12840ttcctgagca cgcagcccgc caacgtgccc
cggggccagg aggactacac caacttcatc 12900agcgcgctgc ggctgatggt ggccgaggtg
ccccagagcg aggtgtacca gtcggggccg 12960gactacttct tccagaccag tcgccagggc
ttgcagaccg tgaacctgag ccaggctttc 13020aagaacttgc agggactgtg gggcgtgcag
gccccggtcg gggaccgcgc gacggtgtcg 13080agcctgctga cgccgaactc gcgcctgctg
ctgctgctgg tggcgccctt cacggacagc 13140ggcagcgtga gccgcgactc gtacctgggc
tacctgctta acctgtaccg cgaggccatc 13200gggcaggcgc acgtggacga gcagacctac
caggagatca cccacgtgag ccgcgcgctg 13260ggccaggagg acccgggcaa cctggaggcc
accctgaact tcctgctgac caaccggtcg 13320cagaagatcc cgccccagta cgcgctgagc
accgaggagg agcgcatcct gcgctacgtg 13380cagcagagcg tggggctgtt cttgatgcag
gagggggcca cgcccagcgc cgcgctcgac 13440atgaccgcgc gcaacatgga gcccagcatg
tacgcccgca accgcccgtt catcaataag 13500ctgatggact acttgcatcg ggcggccgcc
atgaactcgg actactttac caacgccatc 13560ttgaacccgc actggctccc gccgcccggg
ttctacacgg gcgagtacga catgcccgac 13620cccaacgacg ggttcctgtg ggacgacgtg
gacagcagcg tgttctcgcc gcggcccacc 13680accaccaccg tgtggaagaa agagggcggg
gaccggcggc cgtcctcggc gctgtccggt 13740cgcgcgggtg ctgccgcggc ggtgcccgag
gctgccagcc ccttcccgag cctgcccttt 13800tcgctgaaca gcgtgcgcag cagcgagctg
ggtcggctga cgcggccgcg cctgctgggc 13860gaggaggagt acctgaacga ctccttgttg
aagcccgagc gcgagaagaa cttccccaat 13920aacgggatag agagcctggt ggacaagatg
agccgctgga agacgtacgc gcacgagcac 13980agggacgagc cccgagctag cagcgcaggc
acccgtagac gccagcggca cgacaggcag 14040cggggactgg tgtgggacga tgaggattcc
gccgacgaca gcagcgtgtt ggacttgggt 14100gggagtggtg gtggtaaccc gttcgctcac
ctgcgccccc gtatcgggcg cctgatgtaa 14160gaatctgaaa aaataaaaga cggtactcac
caaggccatg gcgaccagcg tgcgttcttc 14220tctgttgttt gtagtagtat gatgaggcgc
gtgtacccgg agggtcctcc tccctcgtac 14280gagagcgtga tgcagcaggc ggtggcggcg
gcgatgcagc ccccgctgga ggcgccttac 14340gtgcccccgc ggtacctggc gcctacggag
gggcggaaca gcattcgtta ctcggagctg 14400gcacccttgt acgataccac ccggttgtac
ctggtggaca acaagtcggc ggacatcgcc 14460tcgctgaact accagaacga ccacagcaac
ttcctgacca ccgtggtgca gaacaacgat 14520ttcaccccca cggaggccag cacccagacc
atcaactttg acgagcgctc gcggtggggc 14580ggccagctga aaaccatcat gcacaccaac
atgcccaacg tgaacgagtt catgtacagc 14640aacaagttca aggcgcgggt gatggtctcg
cgcaagaccc ccaacggggt cacggtaggg 14700gatgattatg atggtagtca ggacgagctg
acctacgagt gggtggagtt tgagctgccc 14760gagggcaact tctcggtgac catgaccatc
gatctgatga acaacgccat catcgacaac 14820tacttggcgg tggggcggca gaacggggtg
ctggagagcg acatcggcgt gaagttcgac 14880acgcgcaact tccggctggg ctgggacccc
gtgaccgagc tggtgatgcc gggcgtgtac 14940accaacgagg ccttccaccc cgacatcgtc
ctgctgcccg gctgcggcgt ggacttcacc 15000gagagccgcc tcagcaacct gctgggcatc
cgcaagcggc agcccttcca ggagggcttc 15060cagatcctgt acgaggacct ggaggggggc
aacatccccg cgctcttgga tgtcgaagcc 15120tatgaagaaa gtaaggaaaa agcagaggct
gaggcaacta cagccgtggc taccgccgcg 15180actgtggcag atgccactgt caccaggggc
gatacattcg ccacccaggc ggaggaagca 15240gccgccctag cggcgaccga tgatagtgaa
agtaagatag tcatcaagcc ggtggagaag 15300gacagcaaga acaggagcta caacgttcta
ccggatggaa agaacaccgc ctaccgcagc 15360tggtacctgg cctacaacta cggcgacccc
gagaagggcg tgcgctcctg gacgctgctc 15420accacctcgg acgtcacctg cggcgtggag
caagtctact ggtcgctgcc cgacatgatg 15480caagacccgg tcaccttccg ctccacgcga
caagttagca actacccggt ggtgggcgcc 15540gagctcctgc ccgtctactc caagagcttc
ttcaacgagc aggccgtcta ctcgcagcag 15600ctgcgtgcct tcacctcgct cacgcacgtc
ttcaaccgct tccccgagaa ccagatcctc 15660gtccgcccgc ccgcgcccac cattaccacc
gtcagtgaaa acgttcctgc tctcacagat 15720cacgggaccc tgccgctgcg cagcagtatc
cggggagtcc agcgcgtgac cgtcactgac 15780gccagacgcc gcacctgccc ctacgtctac
aaggccctgg gcgtagtcgc gccgcgcgtc 15840ctctcgagcc gcaccttcta aaaaatgtcc
attctcatct cgcccagtaa taacaccggt 15900tggggcctgc gcgcgcccag caagatgtac
ggaggcgctc gccaacgctc cacgcaacac 15960cccgtgcgcg tgcgcgggca cttccgcgct
ccctggggcg ccctcaaggg ccgcgtgcgc 16020tcgcgcacca ccgtcgacga cgtgatcgac
caggtggtgg ccgacgcgcg caactacacg 16080cccgccgccg cgcccgcctc caccgtggac
gccgtcatcg acagcgtggt ggccgacgcg 16140cgccggtacg cccgcgccaa gagccggcgg
cggcgcatcg cccggcggca ccggagcacc 16200cccgccatgc gcgcggcgcg agccttgctg
cgcagggcca ggcgcacggg acgcagggcc 16260atgctcaggg cggccagacg cgcggcctcc
ggcagcagca gcgccggcag gacccgcaga 16320cgcgcggcca cggcggcggc ggcggccatc
gccagcatgt cccgcccgcg gcgcggcaac 16380gtgtactggg tgcgcgacgc cgccaccggt
gtgcgcgtgc ccgtgcgcac ccgcccccct 16440cgcacttgaa gatgctgact tcgcgatgtt
gatgtgtccc agcggcgagg aggatgtcca 16500agcgcaaata caaggaagag atgctccagg
tcatcgcgcc tgagatctac ggccccgcgg 16560cggcggtgaa ggaggaaaga aagccccgca
aactgaagcg ggtcaaaaag gacaaaaagg 16620aggaggaaga tgacggactg gtggagtttg
tgcgcgagtt cgccccccgg cggcgcgtgc 16680agtggcgcgg gcggaaagtg aaaccggtgc
tgcggcccgg caccacggtg gtcttcacgc 16740ccggcgagcg ttccggctcc gcctccaagc
gctcctacga cgaggtgtac ggggacgagg 16800acatcctcga gcaggcggtc gagcgtctgg
gcgagtttgc ttacggcaag cgcagccgcc 16860ccgcgccctt gaaagaggag gcggtgtcca
tcccgctgga ccacggcaac cccacgccga 16920gcctgaagcc ggtgaccctg cagcaggtgc
tgccgagcgc ggcgccgcgc cggggcttca 16980agcgcgaggg cggcgaggat ctgtacccga
ccatgcagct gatggtgccc aagcgccaga 17040agctggagga cgtgctggag cacatgaagg
tggaccccga ggtgcagccc gaggtcaagg 17100tgcggcccat caagcaggtg gccccgggcc
tgggcgtgca gaccgtggac atcaagatcc 17160ccacggagcc catggaaacg cagaccgagc
ccgtgaagcc cagcaccagc accatggagg 17220tgcagacgga tccctggatg ccagcggctt
ccaccaccac cactcgccga agacgcaagt 17280acggcgcggc cagcctgctg atgcccaact
acgcgctgca tccttccatc atccccacgc 17340cgggctaccg cggcacgcgc ttctaccgcg
gctacaccag cagccgccgc cgcaagacca 17400ccacccgccg ccgtcgtcgc agccgccgca
gcagcaccgc gacttccgcc ttggtgcgga 17460gagtgtatcg cagcgggcgc gagcctctga
ccctgccgcg cgcgcgctac cacccgagca 17520tcgccattta actaccgcct cctacttgca
gatatggccc tcacatgccg cctccgcgtc 17580cccattacgg gctaccgagg aagaaagccg
cgccgtagaa ggctgacggg gaacgggctg 17640cgtcgccatc accaccggcg gcggcgcgcc
atcagcaagc ggttgggggg aggcttcctg 17700cccgcgctga tccccatcat cgccgcggcg
atcggggcga tccccggcat agcttccgtg 17760gcggtgcagg cctctcagcg ccactgagac
acaaaaaagc atggatttgt aataaaaaaa 17820tggactgacg ctcctggtcc tgtgatgtgt
gtttttagat ggaagacatc aatttttcgt 17880ccctggcacc gcgacacggc acgcggccgt
ttatgggcac ctggagcgac atcggcaaca 17940gccaactgaa cgggggcgcc ttcaattgga
gcagtctctg gagcgggctt aagaatttcg 18000ggtccacgct caaaacctat ggcaacaagg
cgtggaacag cagcacaggg caggcgctga 18060gggaaaagct gaaagagcag aacttccagc
agaaggtggt cgatggcctg gcctcgggca 18120tcaacggggt ggtggacctg gccaaccagg
ccgtgcagaa acagatcaac agccgcctgg 18180acgcggtccc gcccgcgggg tccgtggaga
tgccccaggt ggaggaggag ctgcctcccc 18240tggacaagcg cggcgacaag cgaccgcgtc
ccgacgcgga ggagacgctg ctgacgcaca 18300cggacgagcc gcccccgtac gaggaggcgg
tgaaactggg tctgcccacc acgcggcccg 18360tggcgcctct ggccaccggg gtgctgaaac
ccagcagcag cagccagccc gcgaccctgg 18420acttgcctcc gcctgcttcc cgcccctcca
cagtggctaa gcccctgccg ccggtggccg 18480tcgcgtcgcg cgccccccga ggccgccccc
aggcgaactg gcagagcact ctgaacagca 18540tcgtgggtct gggagtgcag agtgtgaagc
gccgccgctg ctattaaaag acactgtagc 18600gcttaacttg cttgtctgtg tgtgtatatg
tatgtccgcc gaccagaagg aggaagaggc 18660gcgtcgccga gttgcaagat ggccacccca
tcgatgctgc cccagtgggc gtacatgcac 18720atcgccggac aggacgcttc ggagtacctg
agtccgggtc tggtgcagtt cgcccgcgcc 18780acagacacct acttcagtct ggggaacaag
tttaggaacc ccacggtggc gcccacgcac 18840gatgtgacca ccgaccgcag ccagcggctg
acgctgcgct tcgtgcccgt ggaccgcgag 18900gacaacacct actcgtacaa agtgcgctac
acgctggccg tgggcgacaa ccgcgtgctg 18960gacatggcca gcacctactt tgacatccgc
ggcgtgctgg atcggggccc cagcttcaaa 19020ccctactccg gcaccgccta caacagccta
gctcccaagg gagcgcccaa cacctcacag 19080tggaaggatt ccgacagcaa aatgcatact
tttggagttg ctgccatgcc cggtgttgtt 19140ggtaaaaaaa tagaagccga tggtctgcct
attggaatag attcatcctc tggaactgac 19200accataattt atgctgataa aactttccaa
ccagagccac aggttggaag tgacagttgg 19260gtcgacacca atggtgcaga ggaaaaatat
ggaggtagag ctcttaagga cactacaaac 19320atgaagccct gctacggttc ttttgccagg
cctaccaaca aagaaggtgg acaggctaac 19380ataaaagatt ctgaaactgc cagcactact
cctaactatg atatagattt ggcattcttt 19440gacagcaaaa atattgcagc taactacgat
ccagatattg taatgtacac agaaaatgtt 19500gagttgcaaa ctccagatac tcatattgtg
tttaagccag gaacttcaga tgaaagttca 19560gaagccaatt tgggccagca ggccatgccc
aacagaccca actacatcgg gttcagagac 19620aactttatcg ggctcatgta ctacaacagc
actggcaata tgggtgtact ggctggtcag 19680gcctcccagc taaatgctgt ggtggacttg
caggacagaa acaccgaact gtcctaccag 19740ctcttgcttg actctctggg tgacagaacc
aggtatttca gtatgtggaa tcaggcggtg 19800gacagctatg accccgatgt gcgcattatt
gaaaatcacg gtgtggagga tgaactcccc 19860aattattgct tccctttgaa tggtgtaggc
tttacagata cttaccaggg tgttaaagtt 19920aagacagata cagccgctac tggtaccaat
ggaacgcagt gggacaaaga tgataccaca 19980gtcagcactg ccaatgagat ccactcaggc
aatcctttcg ccatggagat caacatccag 20040gccaacctgt ggcggaactt cctctacgcg
aacgtggcgc tgtacctgcc cgactcctac 20100aagtacacgc cggccaacat cacgctgccg
accaacacca acacctacga ttacatgaac 20160ggccgcgtgg tggcgccctc gctggtggac
gcctacatca acatcggggc gcgctggtcg 20220ctggacccca tggacaacgt caaccccttc
aaccaccacc gcaacgcggg cctgcgctac 20280cgctccatgc tcctgggcaa cgggcgctac
gtgcccttcc acatccaggt gccccaaaag 20340tttttcgcca tcaagagcct cctgctcctg
cccgggtcct acacctacga gtggaacttc 20400cgcaaggacg tcaacatgat cctgcagagc
tccctcggca acgacctgcg cacggacggg 20460gcctccatcg ccttcaccag catcaacctc
tacgccacct tcttccccat ggcgcacaac 20520accgcctcca cgctcgaggc catgctgcgc
aacgacacca acgaccagtc cttcaacgac 20580tacctctcgg cggccaacat gctctacccc
atcccggcca acgccaccaa cgtgcccatc 20640tccatcccct cgcgcaactg ggccgccttc
cgcggatggt ccttcacgcg cctcaagacc 20700cgcgagacgc cctcgctcgg ctccgggttc
gacccctact tcgtctactc gggctccatc 20760ccctacctcg acggcacctt ctacctcaac
cacaccttca agaaggtctc catcaccttc 20820gactcctccg tcagctggcc cggcaacgac
cgcctcctga cgcccaacga gttcgaaatc 20880aagcgcaccg tcgacggaga gggatacaac
gtggcccagt gcaacatgac caaggactgg 20940ttcctggtcc agatgctggc ccactacaac
atcggctacc agggcttcta cgtgcccgag 21000ggctacaagg accgcatgta ctccttcttc
cgcaacttcc agcccatgag ccgccaggtc 21060gtggacgagg tcaactacaa ggactaccag
gccgtcaccc tggcctacca gcacaacaac 21120tcgggcttcg tcggctacct cgcgcccacc
atgcgccagg gccagcccta ccccgccaac 21180tacccctacc cgctcatcgg caagagcgcc
gtcgccagcg tcacccagaa aaagttcctc 21240tgcgaccggg tcatgtggcg catccccttc
tccagcaact tcatgtccat gggcgcgctc 21300accgacctcg gccagaacat gctctacgcc
aactccgccc acgcgctaga catgaatttc 21360gaagtcgacc ccatggatga gtccaccctt
ctctatgttg tcttcgaagt cttcgacgtc 21420gtccgagtgc accagcccca ccgcggcgtc
atcgaggccg tctacctgcg cacgcccttc 21480tcggccggca acgccaccac ctaaagcccc
gctcttgctt cttgcaagat gacggcctgt 21540ggctccggcg agcaggagct cagggccatc
ctccgcgacc tgggctgcgg gccctgcttc 21600ctgggcacct tcgacaagcg cttcccggga
ttcatggccc cgcacaagct ggcctgcgcc 21660atcgtcaaca cggccggccg cgagaccggg
ggcgagcact ggctggcctt cgcctggaac 21720ccgcgctccc acacctgcta cctcttcgac
cccttcgggt tctcggacga gcgcctcaag 21780cagatctacc agttcgagta cgagggcctg
ctgcgccgca gcgccctggc caccgaggac 21840cgctgcatca ccctggaaaa gtccacccag
accgtgcagg gtccgcgctc ggccgcctgc 21900gggctcttct gctgcatgtt cctgcacgcc
ttcgtgcact ggcccgaccg ccccatggac 21960aagaacccca ccatgaactt gctgacgggg
gtgcccaacg gcatgctcca gtcgccccag 22020gtggaaccca ccctgcgccg caaccaggag
gcgctctacc gcttcctcaa cgcccactcc 22080gcctactttc gctcccaccg cgcgcgcatc
gagaaggcca ccgccttcga ccgcatgaat 22140caagacatgt aaactgtgtg tatgtgaatg
ctttattcat cataataaac agcacatgtt 22200tatgccacct tctctgaggc tctgacttta
tttagaaatc gaaggggttc tgccggctct 22260cggcgtgccc cgcgggcagg gatacgttgc
ggaactggta cttgggcagc cacttgaact 22320cggggatcag cagcttcggc acggggaggt
cggggaacga gtcgctccac agcttgcgcg 22380tgagttgcag ggcgcccagc aggtcgggcg
cggagatctt gaaatcgcag ttgggacccg 22440cgttctgcgc gcgagagttg cggtacacgg
ggttgcagca ctggaacacc atcagggccg 22500ggtgcttcac gctcgccagc accgtcgcgt
cggtgatgcc ctccacgtcc agatcctcgg 22560cgttggccat cccgaagggg gtcatcttgc
aggtctgccg ccccatgctg ggcacgcagc 22620cgggcttgtg gttgcaatcg cagtgcaggg
ggatcagcat catctgagcc tgctcggagc 22680tcatgcccgg gtacatggcc ttcatgaaag
cctccagctg gcggaaggcc tgctgcgcct 22740tgccgccctc ggtgaagaag accccacagg
acttgctaga gaactggttg gtggcgcagc 22800ccgcgtcgtg cacgcagcag cgcgcgtcgt
tgttggccag ctgcaccacg ctgcgccccc 22860agcggttctg ggtgatcttg gcccggtcgg
ggttctcctt cagcgcgcgc tgcccgttct 22920cgctcgccac atccatctcg atcgtgtgct
ccttctggat catcacggtc ccgtgcaggc 22980accgcagctt gccctcggcc tcggtgcacc
cgtgcagcca cagcgcgcag ccggtgcact 23040cccagttctt gtgggcgatc tgggagtgcg
agtgcacgaa gccctgcagg aagcggccca 23100tcatcgtggt cagggtcttg ttgctggtga
aggtcagcgg gatgccgcgg tgctcctcgt 23160tcacatacag gtggcagatg cggcggtaca
cctcgccctg ctcgggcatc agctggaagg 23220cggacttcag gtcgctctcc acgcggtacc
gctccatcag cagcgtcatc acttccatgc 23280ccttctccca ggccgaaacg atcggcaggc
tcagggggtt cttcaccgtc atcttagtcg 23340ccgccgccga agtcaggggg tcgttctcgt
ccagggtctc aaacactcgc ttgccgtcct 23400tctcggtgat gcgcacgggg ggaaagctga
agcccacggc cgccagctcc tcctcggcct 23460gcctttcgtc ctcgctgtcc tggctgatgt
cttgcaaagg cacatgcttg gtcttgcggg 23520gtttcttttt gggcggcaga ggcggcggcg
gagacgtgct gggcgagcgc gagttctcgc 23580tcaccacgac tatttcttct tcttggccgt
cgtccgagac cacgcggcgg taggcatgcc 23640tcttctgggg cagaggcgga ggcgacgggc
tctcgcggtt cggcgggcgg ctggcagagc 23700cccttccgcg ttcgggggtg cgctcctggc
ggcgctgctc tgactgactt cctccgcggc 23760cggccattgt gttctcctag ggagcaacaa
gcatggagac tcagccatcg tcgccaacat 23820cgccatctgc ccccgccgcc gacgagaacc
agcagcagca gaatgaaagc ttaaccgccc 23880cgccgcccag ccccacctcc gacgccgccg
cggccccaga catgcaagag atggaggaat 23940ccatcgagat tgacctgggc tacgtgacgc
ccgcggagca cgaggaggag ctggcagcgc 24000gcttttcagc cccggaagag aaccaccaag
agcagccaga gcaggaagca gagagcgagc 24060agcagcaggc tgggctcgag catggcgact
acctgagcgg ggcagaggac gtgctcatca 24120agcatctggc ccgccaaagc atcatcgtca
aggacgcgct gctcgaccgc gccgaggtgc 24180ccctcagcgt ggcggagctc agccgcgcct
acgagcgcaa cctcttctcg ccgcgcgtgc 24240cccccaagcg ccagcccaac ggcacctgcg
agcccaaccc gcgcctcaac ttctacccgg 24300tcttcgcggt gcccgaggcc ctggccacct
accacctctt tttcaagaac caaaggatcc 24360ccgtctcctg ccgcgccaac cgcacccgcg
ccgacgccct gctcaacctg ggtcccggcg 24420cccgcctacc tgatatcacc tccttggaag
aggttcccaa gatcttcgag ggtctgggca 24480gcgacgagac tcgggccgcg aacgctctgc
aaggaagcgg agaggagcat gagcaccaca 24540gcgccctggt ggagttggaa ggcgacaacg
cgcgcctggc ggtgctcaag cgcacggtcg 24600agctgaccca cttcgcctac ccggcgctca
acctgccccc caaggtcatg agcgccgtca 24660tggaccaggt gctcatcaag cgcgcctcgc
ccctctcaga ggaggagatg caggaccccg 24720agagctcgga cgagggcaag cccgtggtca
gcgacgagca gctggcgcgc tggctgggag 24780cgagcagcac cccccagagc ctggaagagc
ggcgcaagct catgatggcc gtggtcctgg 24840tgaccgtgga gctggagtgt ctgcgccgct
tcttcgccga cgcggagacc ctgcgcaagg 24900tcgaggagaa cctgcactac ctcttcaggc
acgggttcgt gcgccaggcc tgcaagatct 24960ccaacgtgga gctgaccaac ctggtctcct
acatgggcat cctgcacgag aaccgcctgg 25020ggcagaacgt gctgcacacc accctgcgcg
gggaggcccg ccgcgactac atccgcgact 25080gcgtctacct gtacctctgc cacacctggc
agacgggcat gggcgtgtgg cagcagtgcc 25140tggaggagca gaacctgaaa gagctctgca
agctcctgca gaagaacctc aaggccctgt 25200ggaccgggtt cgacgagcgc accaccgcct
cggacctggc cgacctcatc ttccccgagc 25260gcctgcggct gacgctgcgc aacgggctgc
ccgactttat gagccaaagc atgttgcaaa 25320actttcgctc tttcatcctc gaacgctccg
ggatcctgcc cgccacctgc tccgcactgc 25380cctcggactt cgtgccgctg accttccgcg
agtgcccccc gccgctctgg agccactgct 25440acttgctgcg cctggccaac tacctggcct
accactcgga cgtgatcgag gacgtcagca 25500gcgagggtct gctcgagtgc cactgccgct
gcaacctctg cacgccgcac cgctccttgg 25560cctgcaaccc ccagctgctg agcgagaccc
agatcatcgg caccttcgag ttgcaaggcc 25620ccggcgaggg caaggggggt ctcaaactca
ccccggggct gtggacctcg gcctacttgc 25680gcaagttcgt gcccgaggac taccatccct
tcgagatcag gttctacgag gaccaatccc 25740agccgcccaa ggccgagctg tcggcctgcg
tcatcaccca gggggccatc ctggcccaat 25800tgcaagccat ccagaaatcc cgccaagaat
ttctgctgaa aaagggccac ggggtctact 25860tggaccccca gaccggagag gagctcaacc
ccagcttccc ccaggatgcc ccgaggaagc 25920agcaagaagc tgaaagtgga gctgccgctg
ccgccggagg atttggagga agactgggag 25980agcagtcagg cagaggagat ggaagactgg
gacagcactc aggcagagga ggacagcctg 26040caagacagtc tggaggagga agacgaggtg
gaggaggagg cagaggaaga agcagccgcc 26100gccagaccgt cgtcctcggc ggaggagaaa
gcaagcagca cggataccat ctccgctccg 26160ggtcggggtc gcggcggccg ggcccacagt
agatgggacg agaccgggcg cttcccgaac 26220cccaccaccc agaccggtaa gaaggagcgg
cagggataca agtcctggcg ggggcacaaa 26280aacgccatcg tctcctgctt gcaagcctgc
gggggcaaca tctccttcac ccggcgctac 26340ctgctcttcc accgcggggt gaacttcccc
cgcaacatct tgcattacta ccgtcacctc 26400cacagcccct actactgttt ccaagaagag
gcagaaaccc agcagcagca gcagaaaacc 26460agcggcagca gcagcagcta gaaaatccac
agcggcggca ggtggactga ggatcgcggc 26520gaacgagccg gcgcagaccc gggagctgag
gaaccggatc tttcccaccc tctatgccat 26580cttccagcag agtcgggggc aggagcagga
actgaaagtc aagaaccgtt ctctgcgctc 26640gctcacccgc agttgtctgt atcacaagag
cgaagaccaa cttcagcgca ctctcgagga 26700cgccgaggct ctcttcaaca agtactgcgc
gctcactctt aaagagtagc ccgcgcccgc 26760ccacacacgg aaaaaggcgg gaattacgtc
accacctgcg cccttcgccc gaccatcatc 26820atgagcaaag agattcccac gccttacatg
tggagctacc agccccagat gggcctggcc 26880gccggcgccg cccaggacta ctccacccgc
atgaactggc tcagtgccgg gcccgcgatg 26940atctcacggg tgaatgacat ccgcgcccac
cgaaaccaga tactcctaga acagtcagcg 27000atcaccgcca cgccccgcca tcaccttaat
ccgcgtaatt ggcccgccgc cctggtgtac 27060caggaaattc cccagcccac gaccgtacta
cttccgcgag acgcccaggc cgaagtccag 27120ctgactaact caggtgtcca gctggccggc
ggcgccgccc tgtgtcgtca ccgccccgct 27180cagggtataa agcggctggt gatccgaggc
agaggcacac agctcaacga cgaggtggtg 27240agctcttcgc tgggtctgcg acctgacgga
gtcttccaac tcgccggatc ggggagatct 27300tccttcacgc ctcgtcaggc cgtcctgact
ttggagagtt cgtcctcgca gccccgctcg 27360ggcggcatcg gcactctcca gttcgtggag
gagttcactc cctcggtcta cttcaacccc 27420ttctccggct cccccggcca ctacccggac
gagttcatcc cgaacttcga cgccatcagc 27480gagtcggtgg acggctacga ttgaatgtcc
catggtggcg cggctgacct agctcggctt 27540cgacacctgg accactgtta attaatcgcc
tctcctacga gctcctgcag cagcgccaga 27600agttcacctg cctggtcgga gtcaacccca
tcgtcatcac ccagcagtcg ggcgatacca 27660aggggtgcat ccactgctcc tgcgactccc
ccgactgcgt ccacactctg atcaagaccc 27720tctgcggcct ccgcgacctc ctccccatga
actaatcacc cccttatcca gtgaaataaa 27780gatcatattg atgatgattt tacagaaata
aagatacaat catattgatg atttgagttt 27840aataaaaaat aaagaatcac ttacttgaaa
tctgatacca ggtctctgtc catgttttct 27900gccaacacca cttcactccc ctcttcccag
ctctggtact gcaggccccg gcgggctgca 27960aacttcctcc acacgctgaa ggggatgtca
aattcctcct gtccctcaat cttcatttta 28020tcttctatca gatgtccaaa aagcgcgtcc
gggtggatga tgacttcgac cccgtctacc 28080cctacgatgc agacaacgca ccgaccgtgc
ccttcatcaa cccccccttc gtctcttcag 28140atggattcca agagaagccc ctgggggtgc
tgtccctgcg actggccgac cccgtcacca 28200ccaagaacgg ggaaatcacc ctcaagctgg
gagagggggt ggacctcgac tcctcgggaa 28260aactcatctc caacacggcc accaaggccg
ccgcccctct cagtttttcc aacaacacca 28320tttcccttaa catggatcac cccttttaca
ctaaagatgg aaaattatcc ttacaagttt 28380ctccaccatt aaatatactg agaacaagca
ttctaaacac actagcttta ggttttggat 28440caggtttagg actccgtggc tctgccttgg
cagtacagtt agtctctcca cttacatttg 28500atactgatgg aaacataaag cttaccttag
acagaggttt gcatgttaca acaggagatg 28560caattgaaag caacataagc tgggctaaag
gtttaaaatt tgaagatgga gccatagcaa 28620ccaacattgg aaatgggtta gagtttggaa
gcagtagtac agaaacaggt gttgatgatg 28680cttacccaat ccaagttaaa cttggatctg
gccttagctt tgacagtaca ggagccataa 28740tggctggtaa caaagaagac gataaactca
ctttgtggac aacacctgat ccatcgccaa 28800actgtcaaat actcgcagaa aatgatgcaa
aactaacact ttgcttgact aaatgtggta 28860gtcaaatact ggccactgtg tcagtcttag
ttgtaggaag tggaaaccta aaccccatta 28920ctggcaccgt aagcagtgct caggtgtttc
tacgttttga tgcaaacggt gttcttttaa 28980cagaacattc tacactaaaa aaatactggg
ggtataggca gggagatagc atagatggca 29040ctccatatac caatgctgta ggattcatgc
ccaatttaaa agcttatcca aagtcacaaa 29100gttctactac taaaaataat atagtagggc
aagtatacat gaatggagat gtttcaaaac 29160ctatgcttct cactataacc ctcaatggta
ctgatgacag caacagtaca tattcaatgt 29220cattttcata cacctggact aatggaagct
atgttggagc aacatttggg gctaactctt 29280ataccttctc atacatcgcc caagaatgaa
cactgtatcc caccctgcat gccaaccctt 29340cccaccccac tctgtggaaa aaactctgaa
acacaaaata aaataaagtt caagtgtttt 29400attgattcaa cagttttaca ggattcgagc
agttattttt cctccaccct cccaggacat 29460ggaatacacc accctctccc cccgcacagc
cttgaacatc tgaatgccat tggtgatgga 29520catgcttttg gtctccacgt tccacacagt
ttcagagcga gccagtctcg ggtcggtcag 29580ggagatgaaa ccctccgggc acaattggga
gaagtactcg cctacatggg ggtagagtca 29640taatcgtgca tcaggatagg gcggtggtgc
tgcagcagcg cgcgaataaa ctgctgccgc 29700cgccgctccg tcctgcagga atacaacatg
gcagtggtct cctcagcgat gattcgcacc 29760gcccgcagca taaggcgcct tgtcctccgg
gcacagcagc gcaccctgat ctcacttaaa 29820tcagcacagt aactgcagca cagcaccaca
atattgttca aaatcccaca gtgcaaggcg 29880ctgtatccaa agctcatggc ggggaccaca
gaacccacgt ggccatcata ccacaagcgc 29940aggtagatta agtggcgacc cctcataaac
acgctggaca taaacattac ctcttttggc 30000atgttgtaat tcaccacctc ccggtaccat
ataaacctct gattaaacat ggcgccatcc 30060accaccatcc taaaccagct ggccaaaacc
tgcccgccgg ctatacactg cagggaaccg 30120ggactggaac aatgacagtg gagagcccag
gactcgtaac catggatcat catgctcgtc 30180atgatatcaa tgttggcaca acacaggcac
acgtgcatac acttcctcag gattacaagc 30240tcctcccgcg ttagaaccat atcccaggga
acaacccatt cctgaatcag cgtaaatccc 30300acactgcagg gaagacctcg cacgtaactc
acgttgtgca ttgtcaaagt gttacattcg 30360ggcagcagcg gatgatcctc cagtatggta
gcgcgggttt ctgtctcaaa aggaggtaga 30420cgatccctac tgtacggagt gcgccgagac
aaccgagatc gtgttggtcg tagtgtcatg 30480ccaaatggaa cgccggacgt agtcatattt
cctgaagtct tggcgcgcca aagtctagaa 30540gcggtccata gcttaccgag cggcagcagc
agcggcacac aacaggcgca agagtcagag 30600aaaagactga gctctaacct gtccgcccgc
tctctgctca atatatagcc cagatctaca 30660ctgacgtaaa ggccaaagtc taaaaatacc
cgccaaatag tcacacacgc ccagcacacg 30720cccagaaacc ggtgacacac tcaaaaaaat
acgcgcactt cctcaaacgc ccaaactgcc 30780gtcatttccg ggttcccacg ctacgtcatc
aaaacacgac tttcaaattc cgtcgaccgt 30840taaaaacgtc acccgccccg cccctaacgg
tcgcccgtct ctcagccaat cagcgccccg 30900catccccaaa ttcaaacacc tcatttgcat
attaacgcgc accaaaagtt tgaggtatat 30960tattgatgat g
309711937830DNAPan troglodytes
19catcatcaat aatatacctt attttggatt gaagccaata tgataatgag atgggcggcg
60cggggcggga ggcgggtccg ggggcgggcc ggcgggcggg gcggtgtggc ggaagtggac
120tttgtaagtg tggcggatgt gacttgctag tgccgggcgc ggtaaaagtg acgttttccg
180tgcgcgacaa cgcccacggg aagtgacatt tttcccgcgg tttttaccgg atgttgtagt
240gaatttgggc gtaaccaagt aagatttggc cattttcgcg ggaaaactga aacggggaag
300tgaaatctga ttaatttcgc gttagtcata ccgcgtaata tttgtcgagg gccgagggac
360tttggccgat tacgtggagg actcgcccag gtgttttttg aggtgaattt ccgcgttccg
420ggtcaaagtc tccgttttat tattatagtc agctgacgcg gagtgtattt ataccctctg
480atctcgtcaa gtggccactc ttgagtgcca gcgagtagag ttttctcctc tgccgctctc
540cgctccgctc cgctcggctc tgacaccggg gaaaaaatga gacatttcac ctacgatggc
600ggtgtgctca ccggccagct ggctgctgaa gtcctggaca ccctgatcga ggaggtattg
660gccgataatt atcctccctc gactcctttt gagccaccta cacttcacga actctacgat
720ctggatgtgg tggggcccag cgatccgaac gagcaggcgg tttccagttt ttttccagag
780tccatgttgt tggccagcca ggagggggtc gaacttgaga cccctcctcc gatcgtggat
840tcccccgatc cgccgcagct gactaggcag cccgagcgct gtgcgggacc tgagactatg
900ccccagctgc tacctgaggt gatcgatctc acctgtaatg agtctggttt tccacccagc
960gaggatgagg acgaagaggg tgagcagttt gtgttagatt ctgtggaaca acccgggcga
1020ggatgcaggt cttgtcaata tcaccggaaa aacacaggag actcccagat tatgtgttct
1080ctgtgttata tgaagatgac ctgtatgttt atttacagta agtttatcat ctgtgggcag
1140gtgggctata gtgtgggtgg tggtctttgg ggggtttttt aatatatgtc aggggttatg
1200ctgaagactt ttttattgtg atttttaaag gtccagtgtc tgagcccgag caagaacctg
1260aaccggagcc tgagccttct cgccccagga gaaagcctgt aatcttaact agacccagcg
1320caccggtagc gagaggcctc agcagcgcgg agaccaccga ctccggtgct tcctcatcac
1380ccccggagat tcaccccctg gtgcccctgt gtcccgttaa gcccgttgcc gtgagagtca
1440gtgggcggcg gtctgctgtg gagtgcattg aggacttgct ttttgattca caggaacctt
1500tggacttgag cttgaaacgc cccaggcatt aaacctggtc acctggactg aatgagttga
1560cgcctatgtt tgcttttgaa tgacttaatg tgtatagata ataaagagtg agataatgtt
1620ttaattgcat ggtgtgttta acttgggcgg agtctgctgg gtatataagc ttccctgggc
1680taaacttggt tacacttgac ctcatggagg cctgggagtg tttggagaac tttgccggag
1740ttcgtgcctt gctggacgag agctctaaca atacctcttg gtggtggagg tatttgtggg
1800gctctcccca gggcaagtta gtttgtagaa tcaaggagga ttacaagtgg gaatttgaag
1860agcttttgaa atcctgtggt gagctattgg attctttgaa tctaggccac caggctctct
1920tccaggagaa ggtcatcagg actttggatt tttccacacc ggggcgcatt gcagccgcgg
1980ttgcttttct agcttttttg aaggatagat ggagcgaaga gacccacttg agttcgggct
2040acgtcctgga ttttctggcc atgcaactgt ggagagcatg gatcagacac aagaacaggc
2100tgcaactgtt gtcttccgtc cgcccgttgc tgattccggc ggaggagcaa caggccgggt
2160cagaggaccg ggcccgtcgg gatccggagg agagggcacc gaggccgggc gagaggagcg
2220cgctgaacct gggaaccggg ctgagcggcc atccacatcg ggagtgaatg tcgggcaggt
2280ggtggatctt tttccagaac tgcggcggat tttgactatt agggaggatg ggcaatttgt
2340taagggtctt aagagggaga ggggggcttc tgagcataac gaggaggcca gtaatttagc
2400ttttagcttg atgaccagac accgtccaga gtgcatcact tttcagcaga ttaaggacaa
2460ttgtgccaat gagttggatc tgttgggtca gaagtatagc atagagcagc tgaccactta
2520ctggctgcag ccgggtgatg atctggagga agctattagg gtgtatgcta aggtggccct
2580gcggcccgat tgcaagtaca agctcaaggg gctggtgaat atcaggaatt gttgctacat
2640ttctggcaac ggggcggagg tggagataga gaccgaagac agggtggctt tcagatgcag
2700catgatgaat atgtggccgg gggtgctggg catggacggg gtggtgatta tgaatgtgag
2760gttcacgggg cccaacttta acggcacggt gtttttgggg aacaccaacc tggtcctgca
2820cggggtgagc ttctatgggt ttaacaacac ctgtgtggag gcctggaccg atgtgaaggt
2880ccgcggttgc gccttttatg gatgttggaa ggccatagtg agccgcccta agagcaggag
2940ttccattaag aaatgcttgt ttgagaggtg caccttgggg atcctggccg agggcaactg
3000cagggtgcgc cacaatgtgg cctccgagtg cggttgcttc atgctagtca agagcgtggc
3060ggtaatcaag cataatatgg tgtgcggcaa cagcgaggac aaggcctcac agatgctgac
3120ctgcacggat ggcaactgcc acttgctgaa gaccatccat gtaaccagcc acagccggaa
3180ggcctggccc gtgttcgagc acaacttgct gacccgctgc tccttgcatc tgggcaacag
3240gcggggggtg ttcctgccct atcaatgcaa ctttagtcac accaagatct tgctagagcc
3300cgagagcatg tccaaggtga acttgaacgg ggtgtttgac atgaccatga agatctggaa
3360ggtgctgagg tacgacgaga ccaggtcccg gtgcagaccc tgcgagtgcg ggggcaagca
3420tatgaggaac cagcccgtga tgctggatgt gaccgaggag ctgaggacag accacttggt
3480tctggcctgc accagggccg agtttggttc tagcgatgaa gacacagatt gaggtgggtg
3540agtgggcgtg gcctggggtg gtcatgaaaa tatataagtt gggggtctta gggtctcttt
3600atttgtgttg cagagaccgc cggagccatg agcgggagca gcagcagcag cagtagcagc
3660agcgccttgg atggcagcat cgtgagccct tatttgacga cgcggatgcc ccactgggcc
3720ggggtgcgtc agaatgtgat gggctccagc atcgacggcc gacccgtcct gcccgcaaat
3780tccgccacgc tgacctatgc gaccgtcgcg gggacgccgt tggacgccac cgccgccgcc
3840gccgccaccg cagccgcctc ggccgtgcgc agcctggcca cggactttgc attcctggga
3900ccactggcga caggggctac ttctcgggcc gctgctgccg ccgttcgcga tgacaagctg
3960accgccctgc tggcgcagtt ggatgcgctt actcgggaac tgggtgacct ttctcagcag
4020gtcatggccc tgcgccagca ggtctcctcc ctgcaagctg gcgggaatgc ttctcccaca
4080aatgccgttt aagataaata aaaccagact ctgtttggat taaagaaaag tagcaagtgc
4140attgctctct ttatttcata attttccgcg cgcgataggc cctagaccag cgttctcggt
4200cgttgagggt gcggtgtatc ttctccagga cgtggtagag gtggctctgg acgttgagat
4260acatgggcat gagcccgtcc cgggggtgga ggtagcacca ctgcagagct tcatgctccg
4320gggtggtgtt gtagatgatc cagtcgtagc aggagcgctg ggcatggtgc ctaaaaatgt
4380ccttcagcag caggccgatg gccaggggga ggcccttggt gtaagtgttt acaaaacggt
4440taagttggga agggtgcatt cggggagaga tgatgtgcat cttggactgt atttttagat
4500tggcgatgtt tccgcccaga tcccttctgg gattcatgtt gtgcaggacc accagtacag
4560tgtatccggt gcacttgggg aatttgtcat gcagcttaga gggaaaagcg tggaagaact
4620tggagacgcc tttgtggcct cccagatttt ccatgcattc gtccatgatg atggcaatgg
4680gcccgcggga ggcagcttgg gcaaagatat ttctggggtc gctgacgtcg tagttgtgtt
4740ccagggtgag gtcgtcatag gccattttta caaagcgcgg gcggagggtg cccgactggg
4800ggatgatggt cccctctggc cctggggcgt agttgccctc gcagatctgc atttcccagg
4860ccttaatctc ggagggggga atcatatcca cctgcggggc gatgaagaaa acggtttccg
4920gagccgggga gattaactgg gatgagagca ggtttctaag cagctgtgat tttccacaac
4980cggtgggccc ataaataaca cctataaccg gttgcagctg gtagtttaga gagctgcagc
5040tgccgtcgtc ccggaggagg ggggccacct cgttgagcat gtccctgacg cgcatgttct
5100ccccgaccag atccgccaga aggcgctcgc cgcccaggga cagcagctct tgcaaggaag
5160caaagttttt cagcggcttg aggccgtccg ccgtgggcat gtttttcagg gtctggctca
5220gcagctccag gcggtcccag agctcggtga cgtgctctac ggcatctcta tccagcatat
5280ctcctcgttt cgcgggttgg ggcgactttc gctgtagggc accaagcggt ggtcgtccag
5340cggggccaga gtcatgtcct tccatgggcg cagggtcctc gtcagggtgg tctgggtcac
5400ggtgaagggg tgcgctccgg gctgagcgct tgccaaggtg cgcttgaggc tggttctgct
5460ggtgctgaag cgctgccggt cttcgccctg cgcgtcggcc aggtagcatt tgaccatggt
5520gtcatagtcc agcccctccg cggcgtgtcc cttggcgcgc agcttgccct tggaggtggc
5580gccgcacgag gggcagagca ggctcttgag cgcgtagagc ttgggggcga ggaagaccga
5640ttcgggggag taggcgtccg cgccgcagac cccgcacacg gtctcgcact ccaccagcca
5700ggtgagctcg gggcgcgccg ggtcaaaaac caggtttccc ccatgctttt tgatgcgttt
5760cttacctcgg gtctccatga ggtggtgtcc ccgctcggtg acgaagaggc tgtccgtgtc
5820tccgtagacc gacttgaggg gtcttttctc caggggggtc cctcggtctt cctcgtagag
5880gaactcggac cactctgaga cgaaggcccg cgtccaggcc aggacgaagg aggctatgtg
5940ggaggggtag cggtcgttgt ccactagggg gtccaccttc tccaaggtgt gaagacacat
6000gtcgccttcc tcggcgtcca ggaaggtgat tggcttgtag gtgtaggcca cgtgaccggg
6060ggttcctgac gggggggtat aaaagggggt gggggcgcgc tcgtcgtcac tctcttccgc
6120atcgctgtct gcgagggcca gctgctgggg tgagtattcc ctctcgaagg cgggcatgac
6180ctccgcgctg aggttgtcag tttccaaaaa cgaggaggat ttgatgttca cctgtcccga
6240ggtgatacct ttgagggtac ccgcgtccat ctggtcagaa aacacgatct ttttattgtc
6300cagcttggtg gcgaacgacc cgtagagggc gttggagagc agcttggcga tggagcgcag
6360ggtctggttc ttgtccctgt cggcgcgctc cttggccgcg atgttgagct gcacgtactc
6420gcgcgcgacg cagcgccact cggggaagac ggtggtgcgc tcgtcgggca ccaggcgcac
6480gcgccagccg cggttgtgca gggtgaccag gtccacgctg gtggcgacct cgccgcgcag
6540gcgctcgttg gtccagcaga gacggccgcc cttgcgcgag cagaaggggg gcagggggtc
6600gagctgggtc tcgtccgggg ggtccgcgtc cacggtgaaa accccggggc gcaggcgcgc
6660gtcgaagtag tctatcttgc aaccttgcat gtccagcgcc tgctgccagt cgcgggcggc
6720gagcgcgcgc tcgtaggggt tgagcggcgg gccccagggc atggggtggg tgagtgcgga
6780ggcgtacatg ccgcagatgt catagacgta gaggggctcc cgcaggaccc cgatgtaggt
6840ggggtagcag cggccgccgc ggatgctggc gcgcacgtag tcatacagct cgtgcgaggg
6900ggcgaggagg tcggggccca ggttggtgcg ggcggggcgc tccgcgcgga agacgatctg
6960cctgaagatg gcatgcgagt tggaagagat ggtggggcgc tggaagacgt tgaagctggc
7020gtcctgcagg ccgacggcgt cgcgcacgaa ggaggcgtag gagtcgcgca gcttgtgtac
7080cagctcggcg gtgacctgca cgtcgagcgc gcagtagtcg agggtctcgc ggatgatgtc
7140atatttagcc tgccccttct ttttccacag ctcgcggttg aggacaaact cttcgcggtc
7200tttccagtac tcttggatcg ggaaaccgtc cggttccgaa cggtaagagc ctagcatgta
7260gaactggttg acggcctggt aggcgcagca gcccttctcc acggggaggg cgtaggcctg
7320cgcggccttg cggagcgagg tgtgggtcag ggcgaaggtg tccctgacca tgactttgag
7380gtactggtgc ttgaagtcgg agtcgtcgca gccgccccgc tcccagagcg agaagtcggt
7440gcgcttcttg gagcgggggt tgggcagagc gaaggtgaca tcgttgaaga ggattttgcc
7500cgcgcggggc atgaagttgc gggtgatgcg gaagggcccc ggcacttcag agcggttgtt
7560gatgacctgg gcggcgagca cgatctcgtc gaagccgttg atgttgtggc ccacgatgta
7620gagttccagg aagcggggcc ggccctttac ggtgggcagc ttctttagct cttcgtaggt
7680gagctcctcg ggcgaggcga ggccgtgctc ggccagggcc cagtccgcga ggtgcgggtt
7740gtctctgagg aaggacttcc agaggtcgcg ggccaggagg gtctgcaggc ggtctctgaa
7800ggtcctgaac tggcggccca cggccatttt ttcgggggtg atgcagtaga aggtgagggg
7860gtcttgctgc cagcggtccc agtcgagctg cagggcgagg tcgcgcgcgg cggtgaccag
7920gcgctcgtcg cccccgaatt tcatgaccag catgaagggc acgagctgct ttccgaaggc
7980ccccatccaa gtgtaggtct ctacatcgta ggtgacaaag aggcgctccg tgcgaggatg
8040cgagccgatc gggaagaact ggatctcccg ccaccagttg gaggagtggc tgttgatgtg
8100gtggaagtag aagtcccgtc gccgggccga acactcgtgc tggcttttgt aaaagcgagc
8160gcagtactgg cagcgctgca cgggctgtac ctcatgcacg agatgcacct ttcgcccgcg
8220cacgaggaag ccgaggggaa atctgagccc cccgcctggc tcgcggcatg gctggttctc
8280ttctactttg gatgcgtgtc cgtctccgtc tggctcctcg aggggtgtta cggtggagcg
8340gaccaccacg ccgcgcgagc cgcaggtcca gatatcggcg cgcggcggtc ggagtttgat
8400gacgacatcg cgcagctggg agctgtccat ggtctggagc tcccgcggcg gcggcaggtc
8460agccgggagt tcttgcaggt tcacctcgca gagtcgggcc agggcgcggg gcaggtctag
8520gtggtacctg atctctaggg gcgtgttggt ggcggcgtcg atggcttgca ggagcccgca
8580gccccggggg gcgacgacgg tgccccgcgg ggtggtggtg gtggtggcgg tgcagctcag
8640aagcggtgcc gcgggcgggc ccccggaggt agggggggct ccggtcccgc gggcaggggc
8700ggcagcggca cgtcggcgtg gagcgcgggc aggagttggt gctgtgcccg gaggttgctg
8760gcgaaggcga cgacgcggcg gttgatctcc tggatctggc gcctctgcgt gaagacgacg
8820ggcccggtga gcttgaacct gaaagagagt tcgacagaat caatctcggt gtcattgacc
8880gcggcctggc gcaggatctc ctgcacgtct cccgagttgt cttggtaggc gatctcggcc
8940atgaactgct cgatctcttc ctcctggagg tctccgcgtc cggcgcgttc cacggtggcc
9000gccaggtcgt tggagatgcg ccccatgagc tgcgagaagg cgttgagtcc gccctcgttc
9060cagactcggc tgtagaccac gcccccctgg tcatcgcggg cgcgcatgac cacctgcgcg
9120aggttgagct ccacgtgccg cgcgaagacg gcgtagttgc gcagacgctg gaagaggtag
9180ttgagggtgg tggcggtgtg ctcggccacg aagaagttca tgacccagcg gcgcaacgtg
9240gattcgttga tgtcccccaa ggcctccagc cgttccatgg cctcgtagaa gtccacggcg
9300aagttgaaaa actgggagtt gcgcgccgac acggtcaact cctcctccag aagacggatg
9360agctcggcga cggtgtcgcg cacctcgcgc tcgaaggcta tggggatctc ttcctccgct
9420agcatcacca cctcctcctc ttcctcctct tctggcactt ccatgatggc ttcctcctct
9480tcggggggtg gcggcggcgg cggtggggga gggggcgctc tgcgccggcg gcggcgcacc
9540gggaggcggt ccacgaagcg cgcgatcatc tccccgcggc ggcggcgcat ggtctcggtg
9600acggcgcggc cgttctcccg ggggcgcagt tggaagacgc cgccggacat ctggtgctgg
9660ggcgggtggc cgtgaggcag cgagacggcg ctgacgatgc atctcaacaa ttgctgcgta
9720ggtacgccgc cgagggacct gagggagtcc atatccaccg gatccgaaaa cctttcgagg
9780aaggcgtcta accagtcgca gtcgcaaggt aggctgagca ccgtggcggg cggcgggggg
9840tggggggagt gtctggcgga ggtgctgctg atgatgtaat tgaagtaggc ggacttgaca
9900cggcggatgg tcgacaggag caccatgtcc ttgggtccgg cctgctggat gcggaggcgg
9960tcggctatgc cccaggcttc gttctggcat cggcgcaggt ccttgtagta gtcttgcatg
10020agcctttcca ccggcacctc ttctccttcc tcttctgctt cttccatgtc tgcttcggcc
10080ctggggcggc gccgcgcccc cctgcccccc atgcgcgtga ccccgaaccc cctgagcggt
10140tggagcaggg ccaggtcggc gacgacgcgc tcggccagga tggcctgctg cacctgcgtg
10200agggtggttt ggaagtcatc caagtccacg aagcggtggt aggcgcccgt gttgatggtg
10260taggtgcagt tggccatgac ggaccagttg acggtctggt ggcccggttg cgacatctcg
10320gtgtacctga gtcgcgagta ggcgcgggag tcgaagacgt agtcgttgca agtccgcacc
10380aggtactggt agcccaccag gaagtgcggc ggcggctggc ggtagagggg ccagcgcagg
10440gtggcggggg ctccgggggc caggtcttcc agcatgaggc ggtggtaggc gtagatgtac
10500ctggacatcc aggtgatacc cgcggcggtg gtggaggcgc gcgggaagtc gcgcacccgg
10560ttccagatgt tgcgcagggg cagaaagtgc tccatggtag gcgtgctctg tccagtcaga
10620cgcgcgcagt cgttgatact ctagaccagg gaaaacgaaa gccggtcagc gggcactctt
10680ccgtggtctg gtgaatagat cgcaagggta tcatggcgga gggcctcggt tcgagccccg
10740ggtccgggcc ggacggtccg ccatgatcca cgcggttacc gcccgcgtgt cgaacccagg
10800tgtgcgacgt cagacaacgg tggagtgttc cttttggcgt ttttctggcc gggcgccggc
10860gccgcgtaag agactaagcc gcgaaagcga aagcagtaag tggctcgctc cccgtagccg
10920gagggatcct tgctaagggt tgcgttgcgg cgaaccccgg ttcgaatccc gtactcgggc
10980cggccggacc cgcggctaag gtgttggatt ggcctccccc tcgtataaag accccgcttg
11040cggattgact ccggacacgg ggacgagccc cttttatttt tgctttcccc agatgcatcc
11100ggtgctgcgg cagatgcgcc ccccgcccca gcagcagcaa caacaccagc aagagcggca
11160gcaacagcag cgggagtcat gcagggcccc ctcacccacc ctcggcgggc cggccacctc
11220ggcgtccgcg gccgtgtctg gcgcctgcgg cggcggcggg gggccggctg acgaccccga
11280ggagcccccg cggcgcaggg ccagacacta cctggacctg gaggagggcg agggcctggc
11340gcggctgggg gcgccgtctc ccgagcgcca cccgcgggtg cagctgaagc gcgactcgcg
11400cgaggcgtac gtgcctcggc agaacctgtt cagggaccgc gcgggcgagg agcccgagga
11460gatgcgggac aggaggttca gcgcagggcg ggagctgcgg caggggctga accgcgagcg
11520gctgctgcgc gaggaggact ttgagcccga cgcgcggacg gggatcagcc ccgcgcgcgc
11580gcacgtggcg gccgccgacc tggtgacggc gtacgagcag acggtgaacc aggagatcaa
11640cttccaaaag agtttcaaca accacgtgcg cacgctggtg gcgcgcgagg aggtgaccat
11700cgggctgatg cacctgtggg actttgtaag cgcgctggtg cagaacccca acagcaagcc
11760tctgacggcg cagctgttcc tgatagtgca gcacagcagg gacaacgagg cgtttaggga
11820cgcgctgctg aacatcaccg agcccgaggg tcggtggctg ctggacctga ttaacatcct
11880gcagagcata gtggtgcagg agcgcagcct gagcctggcc gacaaggtgg cggccatcaa
11940ctactcgatg ctgagcctgg gcaagtttta cgcgcgcaag atctaccaga cgccgtacgt
12000gcccatagac aaggaggtga agatcgacgg tttttacatg cgcatggcgc tgaaggtgct
12060caccctgagc gacgacctgg gcgtgtaccg caacgagcgc atccacaagg ccgtgagcgt
12120gagccggcgg cgcgagctga gcgaccgcga gctgatgcac agcctgcagc gggcgctggc
12180gggcgccggc agcggcgaca gggaggcgga gtcctacttc gatgcggggg cggacctgcg
12240ctgggcgccc agccggcggg ccctggaggc cgcgggggtc cgcgaggact atgacgagga
12300cggcgaggag gatgaggagt acgagctaga ggagggcgag tacctggact aaaccgcggg
12360tggtgtttcc ggtagatgca agacccgaac gtggtggacc cggcgctgcg ggcggctctg
12420cagagccagc cgtccggcct taactcctca gacgactggc gacaggtcat ggaccgcatc
12480atgtcgctga cggcgcgtaa cccggacgcg ttccggcagc agccgcaggc caacaggctc
12540tccgccatcc tggaggcggt ggtgcctgcg cgctcgaacc ccacgcacga gaaggtgctg
12600gccatagtga acgcgctggc cgagaacagg gccatccgcc cggacgaggc cgggctggtg
12660tacgacgcgc tgctgcagcg cgtggcccgc tacaacagcg gcaacgtgca gaccaacctg
12720gaccggctgg tgggggacgt gcgcgaggcg gtggcgcagc gcgagcgcgc ggatcggcag
12780ggcaacctgg gctccatggt ggcgctgaat gccttcctga gcacgcagcc ggccaacgtg
12840ccgcgggggc aggaagacta caccaacttt gtgagcgcgc tgcggctgat ggtgaccgag
12900accccccaga gcgaggtgta ccagtcgggc ccggactact tcttccagac cagcagacag
12960ggcctgcaga cggtgaacct gagccaggct ttcaagaacc tgcgggggct gtggggcgtg
13020aaggcgccca ccggcgaccg ggcgacggtg tccagcctgc tgacgcccaa ctcgcgcctg
13080ctgctgctgc tgatcgcgcc gttcacggac agcggcagcg tgtcccggga cacctacctg
13140gggcacctgc tgaccctgta ccgcgaggcc atcgggcagg cgcaggtgga cgagcacacc
13200ttccaggaga tcaccagcgt gagccgcgcg ctggggcagg aggacacgag cagcctggag
13260gcgactctga actacctgct gaccaaccgg cggcagaaga ttccctcgct gcacagcctg
13320acctccgagg aggagcgcat cttgcgctac gtgcagcaga gcgtgagcct gaacctgatg
13380cgcgacgggg tgacgcccag cgtggcgctg gacatgaccg cgcgcaacat ggaaccgggc
13440atgtacgccg cgcaccggcc ttacatcaac cgcctgatgg actacctgca tcgcgcggcg
13500gccgtgaacc ccgagtactt taccaacgcc atcctgaacc cgcactggct cccgccgccc
13560gggttctaca gcgggggctt cgaggtcccg gagaccaacg atggcttcct gtgggacgac
13620atggacgaca gcgtgttctc cccgcggccg caggcgctgg cggaagcgtc cctgctgcgt
13680cccaagaagg aggaggagga ggaggcgagt cgccgccgcg gcagcagcgg cgtggcttct
13740ctgtccgagc tgggggcggc agccgccgcg cgccccgggt ccctgggcgg cagccccttt
13800ccgagcctgg tggggtctct gcacagcgag cgcaccaccc gccctcggct gctgggcgag
13860gacgagtacc tgaataactc cctgctgcag ccggtgcggg agaaaaacct gcctcccgcc
13920ttccccaaca acgggataga gagcctggtg gacaagatga gcagatggaa gacctatgcg
13980caggagcaca gggacgcgcc tgcgctccgg ccgcccacgc ggcgccagcg ccacgaccgg
14040cagcgggggc tggtgtggga tgacgaggac tccgcggacg atagcagcgt gctggacctg
14100ggagggagcg gcaacccgtt cgcgcacctg cgcccccgcc tggggaggat gttttaaaaa
14160aaaaaaaaaa aagcaagaag catgatgcaa aaattaaata aaactcacca aggccatggc
14220gaccgagcgt tggtttcttg tgttcccttc agtatgcggc gcgcggcgat gtaccaggag
14280ggacctcctc cctcttacga gagcgtggtg ggcgcggcgg cggcggcgcc ctcttctccc
14340tttgcgtcgc agctgctgga gccgccgtac gtgcctccgc gctacctgcg gcctacgggg
14400gggagaaaca gcatccgtta ctcggagctg gcgcccctgt tcgacaccac ccgggtgtac
14460ctggtggaca acaagtcggc ggacgtggcc tccctgaact accagaacga ccacagcaat
14520tttttgacca cggtcatcca gaacaatgac tacagcccga gcgaggccag cacccagacc
14580atcaatctgg atgaccggtc gcactggggc ggcgacctga aaaccatcct gcacaccaac
14640atgcccaacg tgaacgagtt catgttcacc aataagttca aggcgcgggt gatggtgtcg
14700cgctcgcaca ccaaggaaga ccgggtggag ctgaagtacg agtgggtgga gttcgagctg
14760ccagagggca actactccga gaccatgacc attgacctga tgaacaacgc gatcgtggag
14820cactatctga aagtgggcag gcagaacggg gtcctggaga gcgacatcgg ggtcaagttc
14880gacaccagga acttccgcct ggggctggac cccgtgaccg ggctggttat gcccggggtg
14940tacaccaacg aggccttcca tcccgacatc atcctgctgc ccggctgcgg ggtggacttc
15000acttacagcc gcctgagcaa cctcctgggc atccgcaagc ggcagccctt ccaggagggc
15060ttcaggatca cctacgagga cctggagggg ggcaacatcc ccgcgctcct cgatgtggag
15120gcctaccagg atagcttgaa ggaaaatgag gcgggacagg aggataccgc ccccgccgcc
15180tccgccgccg ccgagcaggg cgaggatgct gctgacaccg cggccgcgga cggggcagag
15240gccgaccccg ctatggtggt ggaggctccc gagcaggagg aggacatgaa tgacagtgcg
15300gtgcgcggag acaccttcgt cacccggggg gaggaaaagc aagcggaggc cgaggccgcg
15360gccgaggaaa agcaactggc ggcagcagcg gcggcggcgg cgttggccgc ggcggaggct
15420gagtctgagg ggaccaagcc cgccaaggag cccgtgatta agcccctgac cgaagatagc
15480aagaagcgca gttacaacct gctcaaggac agcaccaaca ccgcgtaccg cagctggtac
15540ctggcctaca actacggcga cccgtcgacg ggggtgcgct cctggaccct gctgtgcacg
15600ccggacgtga cctgcggctc ggagcaggtg tactggtcgc tgcccgacat gatgcaagac
15660cccgtgacct tccgctccac gcggcaggtc agcaacttcc cggtggtggg cgccgagctg
15720ctgcccgtgc actccaagag cttctacaac gaccaggccg tctactccca gctcatccgc
15780cagttcacct ctctgaccca cgtgttcaat cgctttcctg agaaccagat tctggcgcgc
15840ccgcccgccc ccaccatcac caccgtcagt gaaaacgttc ctgctctcac agatcacggg
15900acgctaccgc tgcgcaacag catcggagga gtccagcgag tgaccgttac tgacgccaga
15960cgccgcacct gcccctacgt ttacaaggcc ttgggcatag tctcgccgcg cgtcctttcc
16020agccgcactt tttgagcaac accaccatca tgtccatcct gatctcaccc agcaataact
16080ccggctgggg actgctgcgc gcgcccagca agatgttcgg aggggcgagg aagcgttccg
16140agcagcaccc cgtgcgcgtg cgcgggcact tccgcgcccc ctggggagcg cacaaacgcg
16200gccgcgcggg gcgcaccacc gtggacgacg ccatcgactc ggtggtggag caggcgcgca
16260actacaggcc cgcggtctct accgtggacg cggccatcca gaccgtggtg cggggcgcgc
16320ggcggtacgc caagctgaag agccgccgga agcgcgtggc ccgccgccac cgccgccgac
16380ccggggccgc cgccaaacgc gccgccgcgg ccctgcttcg ccgggccaag cgcacgggcc
16440gccgcgccgc catgagggcc gcgcgccgct tggccgccgg catcaccgcc gccaccatgg
16500ccccccgtac ccgaagacgc gcggccgccg ccgccgccgc cgccatcagt gacatggcca
16560gcaggcgccg gggcaacgtg tactgggtgc gcgactcggt gaccggcacg cgcgtgcccg
16620tgcgcttccg ccccccgcgg acttgagatg atgtgaaaaa acaacactga gtctcctgct
16680gttgtgtgta tcccagcggc ggcggcgcgc gcagcgtcat gtccaagcgc aaaatcaaag
16740aagagatgct ccaggtcgtc gcgccggaga tctatgggcc cccgaagaag gaagagcagg
16800attcgaagcc ccgcaagata aagcgggtca aaaagaaaaa gaaagatgat gacgatgccg
16860atggggaggt ggagttcctg cgcgccacgg cgcccaggcg cccggtgcag tggaagggcc
16920ggcgcgtaaa gcgcgtcctg cgccccggca ccgcggtggt cttcacgccc ggcgagcgct
16980ccacccggac tttcaagcgc gtctatgacg aggtgtacgg cgacgaagac ctgctggagc
17040aggccaacga gcgcttcgga gagtttgctt acgggaagcg tcagcgggcg ctggggaagg
17100aggacctgct ggcgctgccg ctggaccagg gcaaccccac ccccagtctg aagcccgtga
17160ccctgcagca ggtgctgccg agcagcgcac cctccgaggc gaagcggggt ctgaagcgcg
17220agggcggcga cctggcgccc accgtgcagc tcatggtgcc caagcggcag aggctggagg
17280atgtgctgga gaaaatgaaa gtagaccccg gtctgcagcc ggacatcagg gtccgcccca
17340tcaagcaggt ggcgccgggc ctcggcgtgc agaccgtgga cgtggtcatc cccaccggca
17400actcccccgc cgccgccacc actaccgctg cctccacgga catggagaca cagaccgatc
17460ccgccgcagc cgcagccgca gccgccgccg cgacctcctc ggcggaggtg cagacggacc
17520cctggctgcc gccggcgatg tcagctcccc gcgcgcgtcg cgggcgcagg aagtacggcg
17580ccgccaacgc gctcctgccc gagtacgcct tgcatccttc catcgcgccc acccccggct
17640accgaggcta tacctaccgc ccgcgaagag ccaagggttc cacccgccgt ccccgccgac
17700gcgccgccgc caccacccgc cgccgccgcc gcagacgcca gcccgcactg gctccagtct
17760ccgtgaggaa agtggcgcgc gacggacaca ccctggtgct gcccagggcg cgctaccacc
17820ccagcatcgt ttaaaagcct gttgtggttc ttgcagatat ggccctcact tgccgcctcc
17880gtttcccggt gccgggatac cgaggaggaa gatcgcgccg caggaggggt ctggccggcc
17940gcggcctgag cggaggcagc cgccgcgcgc accggcggcg acgcgccacc agccgacgca
18000tgcgcggcgg ggtgctgccc ctgttaatcc ccctgatcgc cgcggcgatc ggcgccgtgc
18060ccgggatcgc ctccgtggcc ttgcaagcgt cccagaggca ttgacagact tgcaaacttg
18120caaatatgga aaaaaaaacc ccaataaaaa agtctagact ctcacgctcg cttggtcctg
18180tgactatttt gtagaatgga agacatcaac tttgcgtcgc tggccccgcg tcacggctcg
18240cgcccgttcc tgggacactg gaacgatatc ggcaccagca acatgagcgg tggcgccttc
18300agttggggct ctctgtggag cggcattaaa agtatcgggt ctgccgttaa aaattacggc
18360tcccgggcct ggaacagcag cacgggccag atgttgagag acaagttgaa agagcagaac
18420ttccagcaga aggtggtgga gggcctggcc tccggcatca acggggtggt ggacctggcc
18480aaccaggccg tgcagaataa gatcaacagc agactggacc cccggccgcc ggtggaggag
18540gtgccgccgg cgctggagac ggtgtccccc gatgggcgtg gcgagaagcg cccgcggccc
18600gatagggaag agaccactct ggtcacgcag accgatgagc cgcccccgta tgaggaggcc
18660ctgaagcaag gtctgcccac cacgcggccc atcgcgccca tggccaccgg ggtggtgggc
18720cgccacaccc ccgccacgct ggacttgcct ccgcccgccg atgtgccgca gcagcagaag
18780gcggcacagc cgggcccgcc cgcgaccgcc tcccgttcct ccgccggtcc tctgcgccgc
18840gcggccagcg gcccccgcgg gggggtcgcg aggcacggca actggcagag cacgctgaac
18900agcatcgtgg gtctgggggt gcggtccgtg aagcgccgcc gatgctactg aatagcttag
18960ctaacgtgtt gtatgtgtgt atgcgcccta tgtcgccgcc agaggagctg ctgagtcgcc
19020gccgttcgcg cgcccaccac caccgccact ccgcccctca agatggcgac cccatcgatg
19080atgccgcagt ggtcgtacat gcacatctcg ggccaggacg cctcggagta cctgagcccc
19140gggctggtgc agttcgcccg cgccaccgag agctacttca gcctgagtaa caagtttagg
19200aaccccacgg tggcgcccac gcacgatgtg accaccgacc ggtctcagcg cctgacgctg
19260cggttcattc ccgtggaccg cgaggacacc gcgtactcgt acaaggcgcg gttcaccctg
19320gccgtgggcg acaaccgcgt gctggacatg gcctccacct actttgacat ccgcggggtg
19380ctggaccggg gtcccacttt caagccctac tctggcaccg cctacaactc cctggccccc
19440aagggcgctc ccaactcctg cgagtgggag caagaggaaa ctcaggcagt tgaagaagca
19500gcagaagagg aagaagaaga tgctgacggt caagctgagg aagagcaagc agctaccaaa
19560aagactcatg tatatgctca ggctcccctt tctggcgaaa aaattagtaa agatggtctg
19620caaataggaa cggacgctac agctacagaa caaaaaccta tttatgcaga ccctacattc
19680cagcccgaac cccaaatcgg ggagtcccag tggaatgagg cagatgctac agtcgccggc
19740ggtagagtgc taaagaaatc tactcccatg aaaccatgct atggttccta tgcaagaccc
19800acaaatgcta atggaggtca gggtgtacta acggcaaatg cccagggaca gctagaatct
19860caggttgaaa tgcaattctt ttcaacttct gaaaacgccc gtaacgaggc taacaacatt
19920cagcccaaat tggtgctgta tagtgaggat gtgcacatgg agaccccgga tacgcacctt
19980tcttacaagc ccgcaaaaag cgatgacaat tcaaaaatca tgctgggtca gcagtccatg
20040cccaacagac ctaattacat cggcttcaga gacaacttta tcggcctcat gtattacaat
20100agcactggca acatgggagt gcttgcaggt caggcctctc agttgaatgc agtggtggac
20160ttgcaagaca gaaacacaga actgtcctac cagctcttgc ttgattccat gggtgacaga
20220accagatact tttccatgtg gaatcaggca gtggacagtt atgacccaga tgttagaatt
20280attgaaaatc atggaactga agacgagctc cccaactatt gtttccctct gggtggcata
20340ggggtaactg acacttacca ggctgttaaa accaacaatg gcaataacgg gggccaggtg
20400acttggacaa aagatgaaac ttttgcagat cgcaatgaaa taggggtggg aaacaatttc
20460gctatggaga tcaacctcag tgccaacctg tggagaaact tcctgtactc caacgtggcg
20520ctgtacctac cagacaagct taagtacaac ccctccaatg tggacatctc tgacaacccc
20580aacacctacg attacatgaa caagcgagtg gtggccccgg ggctggtgga ctgctacatc
20640aacctgggcg cgcgctggtc gctggactac atggacaacg tcaacccctt caaccaccac
20700cgcaatgcgg gcctgcgcta ccgctccatg ctcctgggca acgggcgcta cgtgcccttc
20760cacatccagg tgccccagaa gttctttgcc atcaagaacc tcctcctcct gccgggctcc
20820tacacctacg agtggaactt caggaaggat gtcaacatgg tcctccagag ctctctgggt
20880aacgatctca gggtggacgg ggccagcatc aagttcgaga gcatctgcct ctacgccacc
20940ttcttcccca tggcccacaa cacggcctcc acgctcgagg ccatgctcag gaacgacacc
21000aacgaccagt ccttcaatga ctacctctcc gccgccaaca tgctctaccc catacccgcc
21060aacgccacca acgtccccat ctccatcccc tcgcgcaact gggcggcctt ccgcggctgg
21120gccttcaccc gcctcaagac caaggagacc ccctccctgg gctcgggatt cgacccctac
21180tacacctact cgggctccat tccctacctg gacggcacct tctacctcaa ccacactttc
21240aagaaggtct cggtcacctt cgactcctcg gtcagctggc cgggcaacga ccgtctgctc
21300acccccaacg agttcgagat caagcgctcg gtcgacgggg agggctacaa cgtggcccag
21360tgcaacatga ccaaggactg gttcctggtc cagatgctgg ccaactacaa catcggctac
21420cagggcttct acatcccaga gagctacaag gacaggatgt actccttctt caggaacttc
21480cagcccatga gccggcaggt ggtggaccag accaagtaca aggactacca ggaggtgggc
21540atcatccacc agcacaacaa ctcgggcttc gtgggctacc tcgcccccac catgcgcgag
21600ggacaggcct accccgccaa cttcccctat ccgctcatag gcaagaccgc ggtcgacagc
21660atcacccaga aaaagttcct ctgcgaccgc accctctggc gcatcccctt ctccagcaac
21720ttcatgtcca tgggtgcgct ctcggacctg ggccagaact tgctctacgc caactccgcc
21780cacgccctcg acatgacctt cgaggtcgac cccatggacg agcccaccct tctctatgtt
21840ctgttcgaag tctttgacgt ggtccgggtc caccagccgc accgcggcgt catcgagacc
21900gtgtacctgc gtacgccctt ctcggccggc aacgccacca cctaaagaag caagccgcag
21960tcatcgccgc ctgcatgccg tcgggttcca ccgagcaaga gctcagggcc atcgtcagag
22020acctgggatg cgggccctat tttttgggca ccttcgacaa gcgcttccct ggctttgtct
22080ccccacacaa gctggcctgc gccatcgtca acacggccgg ccgcgagacc gggggcgtgc
22140actggctggc cttcgcctgg aacccgcgct ccaaaacatg cttcctcttt gaccccttcg
22200gcttttcgga ccagcggctc aagcaaatct acgagttcga gtacgagggc ttgctgcgtc
22260gcagcgccat cgcctcctcg cccgaccgct gcgtcaccct cgaaaagtcc acccagaccg
22320tgcaggggcc cgactcggcc gcctgcggtc tcttctgctg catgtttctg cacgcctttg
22380tgcactggcc tcagagtccc atggaccgca accccaccat gaacttgctg acgggggtgc
22440ccaactccat gctccagagc ccccaggtcg agcccaccct gcgccgcaac caggagcagc
22500tctacagctt cctggagcgc cactcgcctt acttccgccg ccacagcgca cagatcagga
22560gggccacctc cttctgccac ttgcaagaga tgcaagaagg gtaataacga tgtacacact
22620ttttttctca ataaatggca tctttttatt tatacaagct ctctggggta ttcatttccc
22680accaccaccc gccgttgtcg ccatctggct ctatttagaa atcgaaaggg ttctgccggg
22740agtcgccgtg cgccacgggc agggacacgt tgcgatactg gtagcgggtg ccccacttga
22800actcgggcac caccaggcga ggcagctcgg ggaagttttc gctccacagg ctgcgggtca
22860gcaccagcgc gttcatcagg tcgggcgccg agatcttgaa gtcgcagttg gggccgccgc
22920cctgcgcgcg cgagttgcgg tacaccgggt tgcagcactg gaacaccaac agcgccgggt
22980gcttcacgct ggccagcacg ctgcggtcgg agatcagctc ggcgtccagg tcctccgcgt
23040tgctcagcgc gaacggggtc atcttgggca cttgccgccc caggaagggc gcgtgccccg
23100gtttcgagtt gcagtcgcag cgcagcggga tcagcaggtg cccgtgcccg gactcggcgt
23160tggggtacag cgcgcgcatg aaggcctgca tctggcggaa ggccatctgg gccttggcgc
23220cctccgagaa gaacatgccg caggacttgc ccgagaactg gtttgcgggg cagctggcgt
23280cgtgcaggca gcagcgcgcg tcggtgttgg cgatctgcac cacgttgcgc ccccaccggt
23340tcttcacgat cttggccttg gacgattgct ccttcagcgc gcgctgcccg ttctcgctgg
23400tcacatccat ctcgatcaca tgttccttgt tcaccatgct gctgccgtgc agacacttca
23460gctcgccctc cgtctcggtg cagcggtgct gccacagcgc gcagcccgtg ggctcgaaag
23520acttgtaggt cacctccgcg aaggactgca ggtacccctg caaaaagcgg cccatcatgg
23580tcacgaaggt cttgttgctg ctgaaggtca gctgcagccc gcggtgctcc tcgttcagcc
23640aggtcttgca cacggccgcc agcgcctcca cctggtcggg cagcatcttg aagttcacct
23700tcagctcatt ctccacgtgg tacttgtcca tcagcgtgcg cgccgcctcc atgcccttct
23760cccaggccga caccagcggc aggctcacgg ggttcttcac catcaccgtg gccgccgcct
23820ccgccgcgct ttcgctttcc gccccgctgt tctcttcctc ttcctcctct tcctcgccgc
23880cgcccactcg cagcccccgc accacggggt cgtcttcctg caggcgctgc accttgcgct
23940tgccgttgcg cccctgcttg atgcgcacgg gcgggttgct gaagcccacc atcaccagcg
24000cggcctcttc ttgctcgtcc tcgctgtcca gaatgacctc cggggagggg gggttggtca
24060tcctcagtac cgaggcacgc ttctttttct tcctgggggc gttcgccagc tccgcggctg
24120cggccgctgc cgaggtcgaa ggccgagggc tgggcgtgcg cggcaccagc gcgtcctgcg
24180agccgtcctc gtcctcctcg gactcgagac ggaggcgggc ccgcttcttc gggggcgcgc
24240ggggcggcgg aggcggcggc ggcgacggag acggggacga gacatcgtcc agggtgggtg
24300gacggcgggc cgcgccgcgt ccgcgctcgg gggtggtctc gcgctggtcc tcttcccgac
24360tggccatctc ccactgctcc ttctcctata ggcagaaaga gatcatggag tctctcatgc
24420gagtcgagaa ggaggaggac agcctaaccg ccccctctga gccctccacc accgccgcca
24480ccaccgccaa tgccgccgcg gacgacgcgc ccaccgagac caccgccagt accaccctcc
24540ccagcgacgc acccccgctc gagaatgaag tgctgatcga gcaggacccg ggttttgtga
24600gcggagagga ggatgaggtg gatgagaagg agaaggagga ggtcgccgcc tcagtgccaa
24660aagaggataa aaagcaagac caggacgacg cagataagga tgagacagca gtcgggcggg
24720ggaacggaag ccatgatgct gatgacggct acctagacgt gggagacgac gtgctgctta
24780agcacctgca ccgccagtgc gtcatcgtct gcgacgcgct gcaggagcgc tgcgaagtgc
24840ccctggacgt ggcggaggtc agccgcgcct acgagcggca cctcttcgcg ccgcacgtgc
24900cccccaagcg ccgggagaac ggcacctgcg agcccaaccc gcgtctcaac ttctacccgg
24960tcttcgcggt acccgaggtg ctggccacct accacatctt tttccaaaac tgcaagatcc
25020ccctctcctg ccgcgccaac cgcacccgcg ccgacaaaac cctgaccctg cggcagggcg
25080cccacatacc tgatatcgcc tctctggagg aagtgcccaa gatcttcgag ggtctcggtc
25140gcgacgagaa acgggcggcg aacgctctgc acggagacag cgaaaacgag agtcactcgg
25200gggtgctggt ggagctcgag ggcgacaacg cgcgcctggc cgtactcaag cgcagcatag
25260aggtcaccca ctttgcctac ccggcgctca acctgccccc caaggtcatg agtgtggtca
25320tgggcgagct catcatgcgc cgcgcccagc ccctggccgc ggatgcaaac ttgcaagagt
25380cctccgagga aggcctgccc gcggtcagcg acgagcagct ggcgcgctgg ctggagaccc
25440gcgaccccgc gcagctggag gagcggcgca agctcatgat ggccgcggtg ctggtcaccg
25500tggagctcga gtgtctgcag cgcttcttcg cggaccccga gatgcagcgc aagctcgagg
25560agaccctgca ctacaccttc cgccagggct acgtgcgcca ggcctgcaag atctccaacg
25620tggagctctg caacctggtc tcctacctgg gcatcctgca cgagaaccgc ctcgggcaga
25680acgtcctgca ctccaccctc aaaggggagg cgcgccgcga ctacatccgc gactgcgcct
25740acctcttcct ctgctacacc tggcagacgg ccatgggggt ctggcagcag tgcctggagg
25800agcgcaacct caaggagctg gaaaagctcc tcaagcgcac cctcagggac ctctggacgg
25860gcttcaacga gcgctcggtg gccgccgcgc tggcggacat catctttccc gagcgcctgc
25920tcaagaccct gcagcagggc ctgcccgact tcaccagcca gagcatgctg cagaacttca
25980ggactttcat cctggagcgc tcgggcatcc tgccggccac ttgctgcgcg ctgcccagcg
26040acttcgtgcc catcaagtac agggagtgcc cgccgccgct ctggggccac tgctacctct
26100tccagctggc caactacctc gcctaccact cggacctcat ggaagacgtg agcggcgagg
26160gcctgctcga gtgccactgc cgctgcaacc tctgcacgcc ccaccgctct ctagtctgca
26220acccgcagct gctcagcgag agtcagatta tcggtacctt cgagctgcag ggtccctcgc
26280ctgacgagaa gtccgcggct ccagggctga aactcactcc ggggctgtgg acttccgcct
26340acctacgcaa atttgtacct gaggactacc acgcccacga gatcaggttc tacgaagacc
26400aatcccgccc gcccaaggcg gagctcaccg cctgcgtcat cacccagggg cacatcctgg
26460gccaattgca agccatcaac aaagcccgcc gagagttctt gctgaaaaag ggtcgggggg
26520tgtacctgga cccccagtcc ggcgaggagc taaacccgct acccccgccg ccgccccagc
26580agcgggacct tgcttcccag gatggcaccc agaaagaagc agcagccgcc gccgccgccg
26640cagccataca tgcttctgga ggaagaggag gaggactggg acagtcaggc agaggaggtt
26700tcggacgagg agcaggagga gatgatggaa gactgggagg aggacagcag cctagacgag
26760gaagcttcag aggccgaaga ggtggcagac gcaacaccat cgccctcggt cgcagccccc
26820tcgccggggc ccctgaaatc ctccgaaccc agcaccagcg ctataacctc cgctcctccg
26880gcgccggcgc cacccgcccg cagacccaac cgtagatggg acaccacagg aaccggggtc
26940ggtaagtcca agtgcccgcc gccgccaccg cagcagcagc agcagcagcg ccagggctac
27000cgctcgtggc gcgggcacaa gaacgccata gtcgcctgct tgcaagactg cgggggcaac
27060atctctttcg cccgccgctt cctgctattc caccacgggg tcgcctttcc ccgcaatgtc
27120ctgcattact accgtcatct ctacagcccc tactgcagcg gcgacccaga ggcggcagcg
27180gcagccacag cggcgaccac cacctaggaa gatatcctcc gcgggcaaga cagcggcagc
27240agcggccagg agacccgcgg cagcagcggc gggagcggtg ggcgcactgc gcctctcgcc
27300caacgaaccc ctctcgaccc gggagctcag acacaggatc ttccccactt tgtatgccat
27360cttccaacag agcagaggcc aggagcagga gctgaaaata aaaaacagat ctctgcgctc
27420cctcacccgc agctgtctgt atcacaaaag cgaagatcag cttcggcgca cgctggagga
27480cgcggaggca ctcttcagca aatactgcgc gctcactctt aaagactagc tccgcgccct
27540tctcgaattt aggcgggaga aaactacgtc atcgccggcc gccgcccagc ccgcccagcc
27600gagatgagca aagagattcc cacgccatac atgtggagct accagccgca gatgggactc
27660gcggcgggag cggcccagga ctactccacc cgcatgaact acatgagcgc gggaccccac
27720atgatctcac aggtcaacgg gatccgcgcc cagcgaaacc aaatactgct ggaacaggcg
27780gccatcaccg ccacgccccg ccataatctc aacccccgaa attggcccgc cgccctcgtg
27840taccaggaaa ccccctccgc caccaccgta ctacttccgc gtgacgccca ggccgaagtc
27900cagatgacta actcaggggc gcagctcgcg ggcggctttc gtcacggggc gcggccgctc
27960cgaccaggta taagacacct gatgatcaga ggccgaggta tccagctcaa cgacgagtcg
28020gtgagctctt cgctcggtct ccgtccggac ggaactttcc agctcgccgg atccggccgc
28080tcttcgttca cgccccgcca ggcgtacctg actctgcaga cctcgtcctc ggagccccgc
28140tccggcggca tcggaaccct ccagttcgtg gaggagttcg tgccctcggt ctacttcaac
28200cccttctcgg gacctcccgg acgctacccc gaccagttca ttccgaactt tgacgcggtg
28260aaggactcgg cggacggcta cgactgaatg tcaggtgtcg aggcagagca gcttcgcctg
28320agacacctcg agcactgccg ccgccacaag tgcttcgccc gcggttctgg tgagttctgc
28380tactttcagc tacccgagga gcataccgag gggccggcgc acggcgtccg cctgaccacc
28440cagggcgagg ttacctgttc cctcatccgg gagtttaccc tccgtcccct gctagtggag
28500cgggagcggg gtccctgtgt cctaactatc gcctgcaact gccctaaccc tggattacat
28560caagatcttt gctgtcatct ctgtgctgag tttaataaac gctgagatca gaatctactg
28620gggctcctgt cgccatcctg tgaacgccac cgtcttcacc caccccgacc aggcccaggc
28680gaacctcacc tgcggtctgc atcggagggc caagaagtac ctcacctggt acttcaacgg
28740cacccccttt gtggtttaca acagcttcga cggggacgga gtctccctga aagaccagct
28800ctccggtctc agctactcca tccacaagaa caccaccctc caactcttcc ctccctacct
28860gccgggaacc tacgagtgcg tcaccggccg ctgcacccac ctcacccgcc tgatcgtaaa
28920ccagagcttt ccgggaacag ataactccct cttccccaga acaggaggtg agctcaggaa
28980actccccggg gaccagggcg gagacgtacc ttcgaccctt gtggggttag gattttttat
29040taccgggttg ctggctcttt taatcaaagt ttccttgaga tttgttcttt ccttctacgt
29100gtatgaacac ctcaacctcc aataactcta ccctttcttc ggaatcaggt gacttctctg
29160aaatcgggct tggtgtgctg cttactctgt tgattttttt ccttatcata ctcagccttc
29220tgtgcctcag gctcgccgcc tgctgcgcac acatctatat ctactgctgg ttgctcaagt
29280gcaggggtcg ccacccaaga tgaacaggta catggtccta tcgatcctag gcctgctggc
29340cctggcggcc tgcagcgccg ccaaaaaaga gattaccttt gaggagcccg cttgcaatgt
29400aactttcaag cccgagggtg accaatgcac caccctcgtc aaatgcgtta ccaatcatga
29460gaggctgcgc atcgactaca aaaacaaaac tggccagttt gcggtctata gtgtgtttac
29520gcccggagac ccctctaact actctgtcac cgtcttccag ggcggacagt ctaagatatt
29580caattacact ttcccttttt atgagttatg cgatgcggtc atgtacatgt caaaacagta
29640caacctgtgg cctccctctc cccaggcgtg tgtggaaaat actgggtctt actgctgtat
29700ggctttcgca atcactacgc tcgctctaat ctgcacggtg ctatacataa aattcaggca
29760gaggcgaatc tttatcgatg aaaagaaaat gccttgatcg ctaacaccgg ctttctatct
29820gcagaatgaa tgcaatcacc tccctactaa tcaccaccac cctccttgcg attgcccatg
29880ggttgacacg aatcgaagtg ccagtggggt ccaatgtcac catggtgggc cccgccggca
29940attccaccct catgtgggaa aaatttgtcc gcaatcaatg ggttcatttc tgctctaacc
30000gaatcagtat caagcccaga gccatctgcg atgggcaaaa tctaactctg atcaatgtgc
30060aaatgatgga tgctgggtac tattacgggc agcggggaga aatcattaat tactggcgac
30120cccacaagga ctacatgctg catgtagtcg aggcacttcc cactaccacc cccactacca
30180cctctcccac caccaccacc actactacta ctactactac tactactact actaccacta
30240ccgctgcccg ccatacccgc aaaagcacca tgattagcac aaagccccct cgtgctcact
30300cccacgccgg cgggcccatc ggtgcgacct cagaaaccac cgagctttgc ttctgccaat
30360gcactaacgc cagcgctcat gaactgttcg acctggagaa tgaggatgtc cagcagagct
30420ccgcttgcct gacccaggag gctgtggagc ccgttgccct gaagcagatc ggtgattcaa
30480taattgactc ttcttctttt gccactcccg aataccctcc cgattctact ttccacatca
30540cgggtaccaa agaccctaac ctctctttct acctgatgct gctgctctgt atctctgtgg
30600tctcttccgc gctgatgtta ctggggatgt tctgctgcct gatctgccgc agaaagagaa
30660aagctcgctc tcagggccaa ccactgatgc ccttccccta ccccccggat tttgcagata
30720acaagatatg agctcgctgc tgacactaac cgctttacta gcctgcgctc taacccttgt
30780cgcttgcgac tcgagattcc acaatgtcac agctgtggca ggagaaaatg ttactttcaa
30840ctccacggcc gatacccagt ggtcgtggag tggctcaggt agctacttaa ctatctgcaa
30900tagctccact tcccccggca tatccccaac caagtaccaa tgcaatgcca gcctgttcac
30960cctcatcaac gcttccaccc tggacaatgg actctatgta ggctatgtac cctttggtgg
31020gcaaggaaag acccacgctt acaacctgga agttcgccag cccagaacca ctacccaagc
31080ttctcccacc accaccacca ccaccaccat caccagcagc agcagcagca gcagccacag
31140cagcagcagc agattattga ctttggtttt ggccagctca tctgccgcta cccaggccat
31200ctacagctct gtgcccgaaa ccactcagat ccaccgccca gaaacgacca ccgccaccac
31260cctacacacc tccagcgatc agatgccgac caacatcacc cccttggctc ttcaaatggg
31320acttacaagc cccactccaa aaccagtgga tgcggccgag gtctccgccc tcgtcaatga
31380ctgggcgggg ctgggaatgt ggtggttcgc cataggcatg atggcgctct gcctgcttct
31440gctctggctc atctgctgcc tccaccgcag gcgagccaga ccccccatct atagacccat
31500cattgtcctg aaccccgata atgatgggat ccatagattg gatggcctga aaaacctact
31560tttttctttt acagtatgat aaattgagac atgcctcgca ttttcttgta catgttcctt
31620ctcccacctt ttctggggtg ttctacgctg gccgctgtgt ctcacctgga ggtagactgc
31680ctctcaccct tcactgtcta cctgctttac ggattggtca ccctcactct catctgcagc
31740ctaatcacag taatcatcgc cttcatccag tgcattgatt acatctgtgt gcgcctcgca
31800tacttcagac accacccgca gtaccgagac aggaacattg cccaacttct aagactgctc
31860taatcatgca taagactgtg atctgccttc tgatcctctg catcctgccc accctcacct
31920cctgccagta caccacaaaa tctccgcgca aaagacatgc ctcctgccgc ttcacccaac
31980tgtggaatat acccaaatgc tacaacgaaa agagcgagct ctccgaagct tggctgtatg
32040gggtcatctg tgtcttagtt ttctgcagca ctgtctttgc cctcataatc tacccctact
32100ttgatttggg atggaacgcg atcgatgcca tgaattaccc cacctttccc gcacccgaga
32160taattccact gcgacaagtt gtacccgttg tcgttaatca acgcccccca tcccctacgc
32220ccactgaaat cagctacttt aacctaacag gcggagatga ctgacgccct agatctagaa
32280atggacggca tcagtaccga gcagcgtctc ctagagaggc gcaggcaggc ggctgagcaa
32340gagcgcctca atcaggagct ccgagatctc gttaacctgc accagtgcaa aagaggcatc
32400ttttgtctgg taaagcaggc caaagtcacc tacgagaaga ccggcaacag ccaccgcctc
32460agttacaaat tgcccaccca gcgccagaag ctggtgctca tggtgggtga gaatcccatc
32520accgtcaccc agcactcggt agagaccgag gggtgtctgc actccccctg tcggggtcca
32580gaagacctct gcaccctggt aaagaccctg tgcggtctca gagatttagt cccctttaac
32640taatcaaaca ctggaatcaa taaaaagaat cacttactta aaatcagaca gcaggtctct
32700gtccagttta ttcagcagca cctccttccc ctcctcccaa ctctggtact ccaaacgcct
32760tctggcggca aacttcctcc acaccctgaa gggaatgtca gattcttgct cctgtccctc
32820cgcacccact atcttcatgt tgttgcagat gaagcgcacc aaaacgtctg acgagagctt
32880caaccccgtg tacccctatg acacggaaag cggccctccc tccgtccctt tcctcacccc
32940tcccttcgtg tctcccgatg gattccaaga aagtcccccc ggggtcctgt ctctgaacct
33000ggccgagccc ctggtcactt cccacggcat gctcgccctg aaaatgggaa gtggcctctc
33060cctggacgac gctggcaacc tcacctctca agatatcacc accgctagcc ctcccctcaa
33120aaaaaccaag accaacctca gcctagaaac ctcatccccc ctaactgtga gcacctcagg
33180cgccctcacc gtagcagccg ccgctcccct ggcggtggcc ggcacctccc tcaccatgca
33240atcagaggcc cccctgacag tacaggatgc aaaactcacc ctggccacca aaggccccct
33300gaccgtgtct gaaggcaaac tggccttgca aacatcggcc ccgctgacgg ccgctgacag
33360cagcaccctc acagtcagtg ccacaccacc ccttagcaca agcaatggca gcttgggtat
33420tgacatgcaa gcccccattt acaccaccaa tggaaaacta ggacttaact ttggcgctcc
33480cctgcatgtg gtagacagcc taaatgcact gactgtagtt actggccaag gtcttacgat
33540aaacggaaca gccctacaaa ctagagtctc aggtgccctc aactatgaca catcaggaaa
33600cctagaattg agagctgcag ggggtatgcg agttgatgca aatggtcaac ttatccttga
33660tgtagcttac ccatttgatg cacaaaacaa tctcagcctt aggcttggac agggacccct
33720gtttgttaac tctgcccaca acttggatgt taactacaac agaggcctct acctgttcac
33780atctggaaat accaaaaagc tagaagttaa tatcaaaaca gccaagggtc tcatttatga
33840tgacactgct atagcaatca atgcgggtga tgggctacag tttgactcag gctcagatac
33900aaatccatta aaaactaaac ttggattagg actggattat gactccagca gagccataat
33960tgctaaactg ggaactggcc taagctttga caacacaggt gccatcacag taggcaacaa
34020aaatgatgac aagcttacct tgtggaccac accagaccca tcccctaact gtagaatcta
34080ttcagagaaa gatgctaaat tcacacttgt tttgactaaa tgcggcagtc aggtgttggc
34140cagcgtttct gttttatctg taaaaggtag ccttgcgccc atcagtggca cagtaactag
34200tgctcagatt gtcctcagat ttgatgaaaa tggagttcta ctaagcaatt cttcccttga
34260ccctcaatac tggaactaca gaaaaggtga ccttacagag ggcactgcat ataccaacgc
34320agtgggattt atgcccaacc tcacagcata cccaaaaaca cagagccaaa ctgctaaaag
34380caacattgta agtcaggttt acttgaatgg ggacaaatcc aaacccatga ccctcaccat
34440taccctcaat ggaactaatg aaacaggaga tgccacagta agcacttact ccatgtcatt
34500ctcatggaac tggaatggaa gtaattacat taatgaaacg ttccaaacca actccttcac
34560cttctcctac atcgcccaag aataaaaagc atgacgctgt tgatttgatt caatgtgttt
34620ctgttttatt ttcaagcaca acaaaatcat tcaagtcatt cttccatctt agcttaatag
34680acacagtagc ttaatagacc cagtagtgca aagccccatt ctagcttata gatcagacag
34740tgataattaa ccaccaccac caccatacct tttgattcag gaaatcatga tcatcacagg
34800atcctagtct tcaggccgcc ccctccctcc caagacacag aatacacagt cctctccccc
34860cgactggctt taaataacac catctggttg gtcacagaca tgttcttagg ggttatattc
34920cacacggtct cctgccgcgc caggcgctcg tcggtgatgt tgataaactc tcccggcagc
34980tcgctcaagt tcacgtcgct gtccagcggc tgaacctccg gctgacgcga taactgtgcg
35040accggctgct ggacgaacgg aggccgcgcc tacaaggggg tagagtcata atcctcggtc
35100aggatagggc ggtgatgcag cagcagcgag cgaaacatct gctgccgccg ccgctccgtc
35160cggcaggaaa acaacacgcc ggtggtctcc tccgcgataa tccgcaccgc ccgcagcatc
35220agcttcctcg ttctccgcgc gcagcacctc acccttatct cgctcaaatc ggcgcagtag
35280gtacagcaca gcaccacgat gttattcatg atcccacagt gcagggcgct gtatccaaag
35340ctcatgccgg gaaccaccgc ccccacgtgg ccatcgtacc acaagcgcac gtaaatcaag
35400tgtcgacccc tcatgaacgc gctggacaca aacattactt ccttgggcat gttgtaattc
35460accacctccc ggtaccagat aaacctctgg ttgaacaggg caccttccac caccatcctg
35520aaccaagagg ccagaacctg cccaccggct atgcactgca gggaacccgg gttggaacaa
35580tgacaatgca gactccaagg ctcgtaaccg tggatcatcc ggctgctgaa ggcatcgatg
35640ttggcacaac acagacacac gtgcatgcac tttctcatga ttagcagctc ttccctcgtc
35700aggatcatat cccaaggaat aacccattct tgaatcaacg taaaacccac acagcaggga
35760aggcctcgca cataactcac gttgtgcatg gtcagcgtgt tgcattccgg aaacagcgga
35820tgatcctcca gtatcgaggc gcgggtctcc ttctcacagg gaggtaaagg gtccctgctg
35880tacggactgc gccgggacga ccgagatcgt gttgagcgta gtgtcatgga aaagggaacg
35940ccggacgtgg tcatacttct tgaagcagaa ccaggttcgc gcgtggcagg cctccttgcg
36000tctgcggtct cgccgtctag ctcgctccgt gtgatagttg tagtacagcc actcccgcag
36060agcgtcgagg cgcaccctgg cttccggatc tatgtagact ccgtcttgca ccgcggccct
36120gataatatcc accaccgtag aataagcaac acccagccaa gcaatacact cgctctgcga
36180gcggcagaca ggaggagcgg gcagagatgg gagaaccatg ataaaaaact ttttttaaag
36240aatattttcc aattcttcga aagtaagatc tatcaagtgg cagcgctccc ctccactggc
36300gcggtcaaac tctacggcca aagcacagac aacggcattt ctaagatgtt ccttaatggc
36360gtccaaaaga cacaccgctc tcaagttgca gtaaactatg aatgaaaacc catccggctg
36420attttccaat atagacgcgc cggcagcgtc caccaaaccc agataatttt cttctctcca
36480gcggtttacg atctgtctaa gcaaatccct tatatcaagt ccgaccatgc caaaaatctg
36540ctcaagagcg ccctccacct tcatgtacaa gcagcgcatc atgattgcaa aaattcaggt
36600tcttcagaga cctgtataag attcaaaatg ggaacattaa caaaaattcc tctgtcgcgc
36660agatcccttc gcagggcaag ctgaacataa tcagacaggt ccgaacggac cagtgaggcc
36720aaatccccac caggaaccag atccagagac cctatactga ttatgacgcg catactcggg
36780gctatgctga ccagcgtagc gccgatgtag gcgtgctgca tgggcggcga gataaaatgc
36840aaagtgctgg ttaaaaaatc aggcaaagcc tcgcgcaaaa aagctaacac atcataatca
36900tgctcatgca ggtagttgca ggtaagctca ggaaccaaaa cggaataaca cacgattttc
36960ctctcaaaca tgacttcgcg gatactgcgt aaaacaaaaa attataaata aaaaattaat
37020taaataactt aaacattgga agcctgtctc acaacaggaa aaaccacttt aatcaacata
37080agacgggcca cgggcatgcc ggcatagccg taaaaaaatt ggtccccgtg attaacaagt
37140accacagaca gctccccggt catgtcgggg gtcatcatgt gagactctgt atacacgtct
37200ggattgtgaa catcagacaa acaaagaaat cgagccacgt agcccggagg tataatcacc
37260cgcaggcgga ggtacagcaa aacgaccccc ataggaggaa tcacaaaatt agtaggagaa
37320aaaaatacat aaacaccaga aaaaccctgt tgctgaggca aaatagcgcc ctcccgatcc
37380aaaacaacat aaagcgcttc cacaggagca gccataacaa agacccgagt cttaccagta
37440aaagaaaaaa gatctctcaa cgcagcacca gcaccaacac ttcgcagtgt aaaaggccaa
37500gtgccgagag agtatatata ggaataaaaa gtgacgtaaa cgggcaaagt ccaaaaaacg
37560cccagaaaaa ccgcacgcga acctacgccc cgaaacgaaa gccaaaaaac actagacact
37620cccttccggc gtcaacttcc gctttcccac gctacgtcac ttcccccggt caaacaaact
37680acatatcccg aacttccaag tcgccacgcc caaaacaccg cctacacctc cccgcccgcc
37740ggcccgcccc cggacccgcc tcccgccccg cgccgcccat ctcattatca tattggcttc
37800aatccaaaat aaggtatatt attgatgatg
3783020593PRTPan troglodytes 20Met Arg Arg Ala Ala Met Tyr Gln Glu Gly
Pro Pro Pro Ser Tyr Glu1 5 10
15Ser Val Val Gly Ala Ala Ala Ala Ala Pro Ser Ser Pro Phe Ala Ser
20 25 30Gln Leu Leu Glu Pro Pro
Tyr Val Pro Pro Arg Tyr Leu Arg Pro Thr 35 40
45Gly Gly Arg Asn Ser Ile Arg Tyr Ser Glu Leu Ala Pro Leu
Phe Asp 50 55 60Thr Thr Arg Val Tyr
Leu Val Asp Asn Lys Ser Ala Asp Val Ala Ser65 70
75 80Leu Asn Tyr Gln Asn Asp His Ser Asn Phe
Leu Thr Thr Val Ile Gln 85 90
95Asn Asn Asp Tyr Ser Pro Ser Glu Ala Ser Thr Gln Thr Ile Asn Leu
100 105 110Asp Asp Arg Ser His
Trp Gly Gly Asp Leu Lys Thr Ile Leu His Thr 115
120 125Asn Met Pro Asn Val Asn Glu Phe Met Phe Thr Asn
Lys Phe Lys Ala 130 135 140Arg Val Met
Val Ser Arg Ser His Thr Lys Glu Asp Arg Val Glu Leu145
150 155 160Lys Tyr Glu Trp Val Glu Phe
Glu Leu Pro Glu Gly Asn Tyr Ser Glu 165
170 175Thr Met Thr Ile Asp Leu Met Asn Asn Ala Ile Val
Glu His Tyr Leu 180 185 190Lys
Val Gly Arg Gln Asn Gly Val Leu Glu Ser Asp Ile Gly Val Lys 195
200 205Phe Asp Thr Arg Asn Phe Arg Leu Gly
Leu Asp Pro Val Thr Gly Leu 210 215
220Val Met Pro Gly Val Tyr Thr Asn Glu Ala Phe His Pro Asp Ile Ile225
230 235 240Leu Leu Pro Gly
Cys Gly Val Asp Phe Thr Tyr Ser Arg Leu Ser Asn 245
250 255Leu Leu Gly Ile Arg Lys Arg Gln Pro Phe
Gln Glu Gly Phe Arg Ile 260 265
270Thr Tyr Glu Asp Leu Glu Gly Gly Asn Ile Pro Ala Leu Leu Asp Val
275 280 285Glu Ala Tyr Gln Asp Ser Leu
Lys Glu Asn Glu Ala Gly Gln Glu Asp 290 295
300Thr Ala Pro Ala Ala Ser Ala Ala Ala Glu Gln Gly Glu Asp Ala
Ala305 310 315 320Asp Thr
Ala Ala Ala Asp Gly Ala Glu Ala Asp Pro Ala Met Val Val
325 330 335Glu Ala Pro Glu Gln Glu Glu
Asp Met Asn Asp Ser Ala Val Arg Gly 340 345
350Asp Thr Phe Val Thr Arg Gly Glu Glu Lys Gln Ala Glu Ala
Glu Ala 355 360 365Ala Ala Glu Glu
Lys Gln Leu Ala Ala Ala Ala Ala Ala Ala Ala Leu 370
375 380Ala Ala Ala Glu Ala Glu Ser Glu Gly Thr Lys Pro
Ala Lys Glu Pro385 390 395
400Val Ile Lys Pro Leu Thr Glu Asp Ser Lys Lys Arg Ser Tyr Asn Leu
405 410 415Leu Lys Asp Ser Thr
Asn Thr Ala Tyr Arg Ser Trp Tyr Leu Ala Tyr 420
425 430Asn Tyr Gly Asp Pro Ser Thr Gly Val Arg Ser Trp
Thr Leu Leu Cys 435 440 445Thr Pro
Asp Val Thr Cys Gly Ser Glu Gln Val Tyr Trp Ser Leu Pro 450
455 460Asp Met Met Gln Asp Pro Val Thr Phe Arg Ser
Thr Arg Gln Val Ser465 470 475
480Asn Phe Pro Val Val Gly Ala Glu Leu Leu Pro Val His Ser Lys Ser
485 490 495Phe Tyr Asn Asp
Gln Ala Val Tyr Ser Gln Leu Ile Arg Gln Phe Thr 500
505 510Ser Leu Thr His Val Phe Asn Arg Phe Pro Glu
Asn Gln Ile Leu Ala 515 520 525Arg
Pro Pro Ala Pro Thr Ile Thr Thr Val Ser Glu Asn Val Pro Ala 530
535 540Leu Thr Asp His Gly Thr Leu Pro Leu Arg
Asn Ser Ile Gly Gly Val545 550 555
560Gln Arg Val Thr Val Thr Asp Ala Arg Arg Arg Thr Cys Pro Tyr
Val 565 570 575Tyr Lys Ala
Leu Gly Ile Val Ser Pro Arg Val Leu Ser Ser Arg Thr 580
585 590Phe21964PRTPan troglodytes 21Met Ala Thr
Pro Ser Met Met Pro Gln Trp Ser Tyr Met His Ile Ser1 5
10 15Gly Gln Asp Ala Ser Glu Tyr Leu Ser
Pro Gly Leu Val Gln Phe Ala 20 25
30Arg Ala Thr Asp Ser Tyr Phe Ser Leu Ser Asn Lys Phe Arg Asn Pro
35 40 45Thr Val Ala Pro Thr His Asp
Val Thr Thr Asp Arg Ser Gln Arg Leu 50 55
60Thr Leu Arg Phe Ile Pro Val Asp Arg Glu Asp Thr Ala Tyr Ser Tyr65
70 75 80Lys Ala Arg Phe
Thr Leu Ala Val Gly Asp Asn Arg Val Leu Asp Met 85
90 95Ala Ser Thr Tyr Phe Asp Ile Arg Gly Val
Leu Asp Arg Gly Pro Thr 100 105
110Phe Lys Pro Tyr Ser Gly Thr Ala Tyr Asn Ser Leu Ala Pro Lys Gly
115 120 125Ala Pro Asn Ser Cys Glu Trp
Glu Gln Glu Glu Thr Gln Thr Ala Glu 130 135
140Glu Ala Gln Asp Glu Glu Glu Asp Glu Ala Glu Ala Glu Glu Glu
Met145 150 155 160Pro Gln
Glu Glu Gln Ala Pro Val Lys Lys Thr His Val Tyr Ala Gln
165 170 175Ala Pro Leu Ser Gly Glu Lys
Ile Thr Lys Asp Gly Leu Gln Ile Gly 180 185
190Thr Asp Ala Thr Ala Thr Glu Gln Lys Pro Ile Tyr Ala Asp
Pro Thr 195 200 205Phe Gln Pro Glu
Pro Gln Ile Gly Glu Ser Gln Trp Asn Glu Ala Asp 210
215 220Ala Ser Val Ala Gly Gly Arg Val Leu Lys Lys Thr
Thr Pro Met Lys225 230 235
240Pro Cys Tyr Gly Ser Tyr Ala Arg Pro Thr Asn Ala Asn Gly Gly Gln
245 250 255Gly Val Leu Val Glu
Lys Asp Gly Gly Lys Met Glu Ser Gln Val Asp 260
265 270Met Gln Phe Phe Ser Thr Ser Glu Asn Ala Arg Asn
Glu Ala Asn Asn 275 280 285Ile Gln
Pro Lys Leu Val Leu Tyr Ser Glu Asp Val His Met Glu Thr 290
295 300Pro Asp Thr His Ile Ser Tyr Lys Pro Ala Lys
Ser Asp Asp Asn Ser305 310 315
320Lys Val Met Leu Gly Gln Gln Ser Met Pro Asn Arg Pro Asn Tyr Ile
325 330 335Gly Phe Arg Asp
Asn Phe Ile Gly Leu Met Tyr Tyr Asn Ser Thr Gly 340
345 350Asn Met Gly Val Leu Ala Gly Gln Ala Ser Gln
Leu Asn Ala Val Val 355 360 365Asp
Leu Gln Asp Arg Asn Thr Glu Leu Ser Tyr Gln Leu Leu Leu Asp 370
375 380Ser Met Gly Asp Arg Thr Arg Tyr Phe Ser
Met Trp Asn Gln Ala Val385 390 395
400Asp Ser Tyr Asp Pro Asp Val Arg Ile Ile Glu Asn His Gly Thr
Glu 405 410 415Asp Glu Leu
Pro Asn Tyr Cys Phe Pro Leu Gly Gly Ile Gly Val Thr 420
425 430Asp Thr Tyr Gln Ala Ile Lys Thr Asn Gly
Asn Gly Asn Gly Gly Gly 435 440
445Asn Thr Thr Trp Thr Lys Asp Glu Thr Phe Ala Asp Arg Asn Glu Ile 450
455 460Gly Val Gly Asn Asn Phe Ala Met
Glu Ile Asn Leu Ser Ala Asn Leu465 470
475 480Trp Arg Asn Phe Leu Tyr Ser Asn Val Ala Leu Tyr
Leu Pro Asp Lys 485 490
495Leu Lys Tyr Asn Pro Ser Asn Val Glu Ile Ser Asp Asn Pro Asn Thr
500 505 510Tyr Asp Tyr Met Asn Lys
Arg Val Val Ala Pro Gly Leu Val Asp Cys 515 520
525Tyr Ile Asn Leu Gly Ala Arg Trp Ser Leu Asp Tyr Met Asp
Asn Val 530 535 540Asn Pro Phe Asn His
His Arg Asn Ala Gly Leu Arg Tyr Arg Ser Met545 550
555 560Leu Leu Gly Asn Gly Arg Tyr Val Pro Phe
His Ile Gln Val Pro Gln 565 570
575Lys Phe Phe Ala Ile Lys Asn Leu Leu Leu Leu Pro Gly Ser Tyr Thr
580 585 590Tyr Glu Trp Asn Phe
Arg Lys Asp Val Asn Met Val Leu Gln Ser Ser 595
600 605Leu Gly Asn Asp Leu Arg Val Asp Gly Ala Ser Ile
Lys Phe Glu Ser 610 615 620Ile Cys Leu
Tyr Ala Thr Phe Phe Pro Met Ala His Asn Thr Ala Ser625
630 635 640Thr Leu Glu Ala Met Leu Arg
Asn Asp Thr Asn Asp Gln Ser Phe Asn 645
650 655Asp Tyr Leu Ser Ala Ala Asn Met Leu Tyr Pro Ile
Pro Ala Asn Ala 660 665 670Thr
Asn Val Pro Ile Ser Ile Pro Ser Arg Asn Trp Ala Ala Phe Arg 675
680 685Gly Trp Ala Phe Thr Arg Leu Lys Thr
Lys Glu Thr Pro Ser Leu Gly 690 695
700Ser Gly Phe Asp Pro Tyr Tyr Thr Tyr Ser Gly Ser Ile Pro Tyr Leu705
710 715 720Asp Gly Thr Phe
Tyr Leu Asn His Thr Phe Lys Lys Val Ser Val Thr 725
730 735Phe Asp Ser Ser Val Ser Trp Pro Gly Asn
Asp Arg Leu Leu Thr Pro 740 745
750Asn Glu Phe Glu Ile Lys Arg Ser Val Asp Gly Glu Gly Tyr Asn Val
755 760 765Ala Gln Cys Asn Met Thr Lys
Asp Trp Phe Leu Ile Gln Met Leu Ala 770 775
780Asn Tyr Asn Ile Gly Tyr Gln Gly Phe Tyr Ile Pro Glu Ser Tyr
Lys785 790 795 800Asp Arg
Met Tyr Ser Phe Phe Arg Asn Phe Gln Pro Met Ser Arg Gln
805 810 815Val Val Asp Glu Thr Lys Tyr
Lys Asp Tyr Gln Gln Val Gly Ile Ile 820 825
830His Gln His Asn Asn Ser Gly Phe Val Gly Tyr Leu Ala Pro
Thr Met 835 840 845Arg Glu Gly Gln
Ala Tyr Pro Ala Asn Phe Pro Tyr Pro Leu Ile Gly 850
855 860Lys Thr Ala Val Asp Ser Val Thr Gln Lys Lys Phe
Leu Cys Asp Arg865 870 875
880Thr Leu Trp Arg Ile Pro Phe Ser Ser Asn Phe Met Ser Met Gly Ala
885 890 895Leu Thr Asp Leu Gly
Gln Asn Leu Leu Tyr Ala Asn Ser Ala His Ala 900
905 910Leu Asp Met Thr Phe Glu Val Asp Pro Met Asp Glu
Pro Thr Leu Leu 915 920 925Tyr Val
Leu Phe Glu Val Phe Asp Val Val Arg Val His Gln Pro His 930
935 940Arg Gly Val Ile Glu Thr Val Tyr Leu Arg Thr
Pro Phe Ser Ala Gly945 950 955
960Asn Ala Thr Thr22578PRTPan troglodytes 22Met Lys Arg Thr Lys Thr
Ser Asp Glu Ser Phe Asn Pro Val Tyr Pro1 5
10 15Tyr Asp Thr Glu Ser Gly Pro Pro Ser Val Pro Phe
Leu Thr Pro Pro 20 25 30Phe
Val Ser Pro Asp Gly Phe Gln Glu Ser Pro Pro Gly Val Leu Ser 35
40 45Leu Asn Leu Ala Glu Pro Leu Val Thr
Ser His Gly Met Leu Ala Leu 50 55
60Lys Met Gly Ser Gly Leu Ser Leu Asp Asp Ala Gly Asn Leu Thr Ser65
70 75 80Gln Asp Ile Thr Thr
Ala Ser Pro Pro Leu Lys Lys Thr Lys Thr Asn 85
90 95Leu Ser Leu Glu Thr Ser Ser Pro Leu Thr Val
Ser Thr Ser Gly Ala 100 105
110Leu Thr Val Ala Ala Ala Ala Pro Leu Ala Val Ala Gly Thr Ser Leu
115 120 125Thr Met Gln Ser Glu Ala Pro
Leu Thr Val Gln Asp Ala Lys Leu Thr 130 135
140Leu Ala Thr Lys Gly Pro Leu Thr Val Ser Glu Gly Lys Leu Ala
Leu145 150 155 160Gln Thr
Ser Ala Pro Leu Thr Ala Ala Asp Ser Ser Thr Leu Thr Val
165 170 175Ser Ala Thr Pro Pro Leu Ser
Thr Ser Asn Gly Ser Leu Gly Ile Asp 180 185
190Met Gln Ala Pro Ile Tyr Thr Thr Asn Gly Lys Leu Gly Leu
Asn Phe 195 200 205Gly Ala Pro Leu
His Val Val Asp Ser Leu Asn Ala Leu Thr Val Val 210
215 220Thr Gly Gln Gly Leu Thr Ile Asn Gly Thr Ala Leu
Gln Thr Arg Val225 230 235
240Ser Gly Ala Leu Asn Tyr Asp Thr Ser Gly Asn Leu Glu Leu Arg Ala
245 250 255Ala Gly Gly Met Arg
Val Asp Ala Asn Gly Gln Leu Ile Leu Asp Val 260
265 270Ala Tyr Pro Phe Asp Ala Gln Asn Asn Leu Ser Leu
Arg Leu Gly Gln 275 280 285Gly Pro
Leu Phe Val Asn Ser Ala His Asn Leu Asp Val Asn Tyr Asn 290
295 300Arg Gly Leu Tyr Leu Phe Thr Ser Gly Asn Thr
Lys Lys Leu Glu Val305 310 315
320Asn Ile Lys Thr Ala Lys Gly Leu Ile Tyr Asp Asp Thr Ala Ile Ala
325 330 335Ile Asn Ala Gly
Asp Gly Leu Gln Phe Asp Ser Gly Ser Asp Thr Asn 340
345 350Pro Leu Lys Thr Lys Leu Gly Leu Gly Leu Asp
Tyr Asp Ser Ser Arg 355 360 365Ala
Ile Ile Ala Lys Leu Gly Thr Gly Leu Ser Phe Asp Asn Thr Gly 370
375 380Ala Ile Thr Val Gly Asn Lys Asn Asp Asp
Lys Leu Thr Leu Trp Thr385 390 395
400Thr Pro Asp Pro Ser Pro Asn Cys Arg Ile Tyr Ser Glu Lys Asp
Ala 405 410 415Lys Phe Thr
Leu Val Leu Thr Lys Cys Gly Ser Gln Val Leu Ala Ser 420
425 430Val Ser Val Leu Ser Val Lys Gly Ser Leu
Ala Pro Ile Ser Gly Thr 435 440
445Val Thr Ser Ala Gln Ile Val Leu Arg Phe Asp Glu Asn Gly Val Leu 450
455 460Leu Ser Asn Ser Ser Leu Asp Pro
Gln Tyr Trp Asn Tyr Arg Lys Gly465 470
475 480Asp Leu Thr Glu Gly Thr Ala Tyr Thr Asn Ala Val
Gly Phe Met Pro 485 490
495Asn Leu Thr Ala Tyr Pro Lys Thr Gln Ser Gln Thr Ala Lys Ser Asn
500 505 510Ile Val Ser Gln Val Tyr
Leu Asn Gly Asp Lys Ser Lys Pro Met Thr 515 520
525Leu Thr Ile Thr Leu Asn Gly Thr Asn Glu Thr Gly Asp Ala
Thr Val 530 535 540Ser Thr Tyr Ser Met
Ser Phe Ser Trp Asn Trp Asn Gly Ser Asn Tyr545 550
555 560Ile Asn Glu Thr Phe Gln Thr Asn Ser Phe
Thr Phe Ser Tyr Ile Ala 565 570
575Gln Glu
User Contributions:
Comment about this patent or add new information about this topic: