Patent application title: Increased Production of Isobutanol in Yeast with Reduced Mitochondrial Amino Acid Biosynthesis
Inventors:
IPC8 Class: AC12P716FI
USPC Class:
1 1
Class name:
Publication date: 2016-10-20
Patent application number: 20160304909
Abstract:
Yeast cells with reduced activity of certain enzymes involved in branched
chain amino acid biosynthesis in yeast mitochondria are described. Target
enzymes include threonine deaminase, isopropylmalate synthase, and
optionally branched chain amino acid transaminase.Claims:
1-15. (canceled)
16. A recombinant yeast host cell which produces isobutanol and comprises mitochondria which is substantially devoid of branched chain amino acid transaminase activity.
17. The recombinant yeast host cell of claim 16, wherein the mitochondria is further substantially devoid of isopropylmalate synthase activity.
18. The recombinant yeast host cell of claim 17, wherein the mitochondria is further substantially devoid of threonine deaminase activity.
19. The recombinant yeast cell of claim 16, wherein the branched chain amino acid transaminase activity is defined by the enzyme classification number EC 2.6.1.42.
20. The recombinant yeast cell of claim 17, wherein the branched chain amino acid transaminase activity is defined by the enzyme classification number EC 2.6.1.42.
21. The recombinant yeast cell of claim 18, wherein the branched chain amino acid transaminase activity is defined by the enzyme classification number EC 2.6.1.42.
22. The recombinant yeast host cell of claim 17, wherein the isopropylmalate synthase activity is defined by the enzyme classification number EC 2.3.3.13.
23. The recombinant yeast host cell of claim 18, wherein the isopropylmalate synthase activity is defined by the enzyme classification number EC 2.3.3.13.
24. The recombinant yeast host cell of claim 18, wherein the threonine deaminase activity is defined by the enzyme classification number EC 4.3.1.19
25. The recombinant yeast cell of claim 16, wherein the yeast cell comprises a disruption in a BAT1 gene.
26. The recombinant yeast cell of claim 17, wherein the yeast cell comprises a disruption in a LEU4 gene.
27. The recombinant yeast cell of claim 18, wherein the yeast cell comprises a disruption in a ILV1 gene.
28. The recombinant yeast cell of claim 18, wherein the yeast cell comprises disruptions in a BAT1 gene, a LEU4 gene, and an ILV1 gene.
29. The recombinant yeast cell of claim 16, wherein the recombinant yeast cell further comprises reduced or eliminated endogenous pyruvate decarboxylase activity.
30. The recombinant yeast cell of claim 17, wherein the recombinant yeast cell further comprises reduced or eliminated endogenous pyruvate decarboxylase activity.
31. The recombinant yeast cell of claim 18, wherein the recombinant yeast cell further comprises reduced or eliminated endogenous pyruvate decarboxylase activity.
32. The recombinant yeast cell of claim 16, wherein the yeast is selected from the group consisting of Saccharomyces, Schizosaccharomyces, Hansuela, Candida, Kluveromyces, Yarrowia, and Pichia.
33. A method of producing isobutanol comprising growing a recombinant yeast cell of claim 16, wherein the recombinant yeast cell is grown under conditions wherein isobutanol is produced.
34. A method of producing isobutanol comprising growing a recombinant yeast cell of claim 17, wherein the recombinant yeast cell is grown under conditions wherein isobutanol is produced.
35. A method of producing isobutanol comprising growing a recombinant yeast cell of claim 18, wherein the recombinant yeast cell is grown under conditions wherein isobutanol is produced.
Description:
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional of and claims the benefit of U.S. application Ser. No. 14/302,097, filed Jun. 11, 2014, which is a divisional and claims the benefit of U.S. application Ser. No. 13/889,999, filed on May 8, 2013, now U.S. Pat. No. 8,785,166, which is a divisional of and claims the benefit of U.S. application Ser. No. 12/617,039, filed Nov. 12, 2009, now U.S. Pat. No. 8,465,964, which is related to and claims the benefit of priority of U.S. Provisional Application No. 61/114,072, filed Nov. 13, 2008. Each of the referenced applications is herein incorporated by reference.
FIELD OF THE INVENTION
[0002] The invention relates to the field of microbiology. More specifically, recombinant yeast strains are disclosed that have reduced amino acid biosynthesis and increased isobutanol production.
BACKGROUND OF THE INVENTION
[0003] Butanol is an important industrial chemical, useful as a fuel additive, as a feedstock chemical in the plastics industry, and as a foodgrade extractant in the food and flavor industry. Each year 10 to 12 billion pounds of butanol are produced by petrochemical means and the need for this commodity chemical will likely increase.
[0004] Isobutanol is produced biologically as a by-product of yeast fermentation. It is a component of "fusel oil" that forms as a result of incomplete metabolism of amino acids by this group of fungi. Isobutanol is specifically produced from catabolism of L-valine in the cytoplasm. After the amine group of L-valine is harvested as a nitrogen source, the resulting .alpha.-keto acid is decarboxylated and reduced to isobutanol by enzymes of the so-called Ehrlich pathway (Dickinson et al., J. Biol. Chem. 273(40):25752-25756 (1998)). Yields of fusel oil and/or its components achieved during beverage fermentation are typically low. For example, the concentration of isobutanol produced in beer fermentation is reported to be less than 16 parts per million (Garcia et al., Process Biochemistry 29:303-309 (1994)). Addition of exogenous L-valine to the fermentation increases the yield of isobutanol, as described by Dickinson et al., supra, wherein it is reported that a yield of isobutanol of 3 g/L is obtained by providing L-valine at a concentration of 20 g/L in the fermentation. However, the use of valine as a feed-stock would be cost prohibitive for industrial scale isobutanol production.
[0005] Additionally Van Nedervelde et al (Proceedings of the Congress--European Brewery Convention (2003), 29th, 50/1-50/10) have demonstrated the deletions in the gene encoding the BAT 1 mitochondrial protein in yeast result in strains having increased levels of higher alcohols. Similarly Nako et al (WO 2007032522) note that amyl alcohol and/or isobutanol and/or isoamyl acetate levels in yeast used for the production of alcoholic beverages may be altered via manipulation of the BAT1 and BAT2 genes. The art is silent with respect to the down regulation of other genes encoding proteins that are functional in the mitochondria for the enhanced production of isobutanol in yeast.
[0006] There is a need for attaining higher amounts of isobutanol through yeast fermentation without addition of valine or other isobutanol production intermediates.
SUMMARY OF THE INVENTION
[0007] Provided herein are recombinant yeast host cells which comprise mitochondria which are substantially devoid of an enzyme activity selected from the group consisting of threonine deaminase and isopropylmalate synthase activity. In some embodiments, the host cells produce isobutanol. In some embodiments, the mitochondria is substantially devoid of branched chain amino acid transaminase activity, and in some embodiments, the mitochondria is substantially devoid of pyruvate dehydrogenase activity. In some embodiments, endogenous pyruvate decarboxylase activity is reduced.
[0008] In some embodiments, the threonine deaminase activity is defined by the enzyme classification number EC 4.3.1.19 and the isopropylmalate synthase activity is defined by the enzyme classification number EC 2.3.3.13. In some embodiments, the branched chain amino acid transaminase activity is defined by the enzyme classification number EC 2.6.1.42. In some embodiments, the pyruvate dehydrogenase activity is defined by the enzyme classification number EC 1.2.4.1. In some embodiments, the pyruvate dehydrogenase activity is defined by a multienzyme complex comprising proteins selected from the group consisting of: PDA1, PDA1, PDB1, LAT1, LPD1, and PDX1.
[0009] In some embodiments, the yeast is selected from the group consisting of Saccharomyces, Schizosaccharomyces, Hansenula, Candida, Kluyveromyces, Yarrowia and Pichia.
[0010] In some embodiments, yeast cells provided comprise a disruption in a gene selected from the group consisting of ILV1 and LEU4. In some embodiments, yeast cells provided comprise a disruption in the BAT1 gene. In some embodiments, yeast cells provided comprise a disruption in a gene encoding a protein selected from the group consisting of PDA1, PDA1, PDB1, LAT1, LPD1, and PDX1
[0011] In some embodiments, yeast cells provided herein are Saccharomyces and wherein; a) the ILV1 gene encodes a polypeptide having at least 80% identity to an amino acid sequence as set forth in SEQ ID NO: 2, based on the Clustal W method of alignment using the default parameters of GAP PENALTY=10, GAP LENGTH PENALTY=0.1, and Gonnet 250 series of protein weight matrix.; and b) the LEU4 gene encodes a polypeptide having at least 80% identity to the amino acid sequence set forth in SEQ ID NO: 28 based on the Clustal W method of alignment using the default parameters of GAP PENALTY=10, GAP LENGTH PENALTY=0.1, and Gonnet 250 series of protein weight matrix.
[0012] In some embodiments, yeast cells provided herein are Saccharomyces and wherein the BAT1 gene encodes a protein having at least 80% identity to the amino acid sequence as set forth in SEQ ID NO:16 based on the Clustal W method of alignment using the default parameters of GAP PENALTY=10, GAP LENGTH PENALTY=0.1, and Gonnet 250 series of protein weight matrix.
[0013] In some embodiments, yeast cells provided herein are Saccharomyces and wherein; a) the PDA1 polypeptide has at least 80% identity to the amino acid sequence as set forth in SEQ ID NO:70; b) the PDB1 polypeptide has at least 80% identity to the amino acid sequence as set forth in SEQ ID NO: 58; c) the LAT1 polypeptide has at least 80% identity to the amino acid sequence as set forth in SEQ ID NO: 106; d) the LPD1 polypeptide has at least 80% identity to the amino acid sequence as set forth in SEQ ID NO: 108; and e) the PDX1 polypeptide has at least 80% identity to the amino acid sequence as set forth in SEQ ID NO: 110; wherein identity of polypeptides recited in parts (a)-(e) is based on the Clustal W method of alignment using the default parameters of GAP PENALTY=10, GAP LENGTH PENALTY=0.1, and Gonnet 250 series of protein weight matrix.
[0014] Also provided herein are methods for the production of isobutanol comprising growing provided host cells under conditions wherein isobutanol is produced.
BRIEF DESCRIPTION OF THE FIGURES AND SEQUENCE DESCRIPTIONS
[0015] The invention can be more fully understood from the following detailed description, figures, and the accompanying sequence descriptions, which form a part of this application.
[0016] FIG. 1 shows biosynthetic pathways for amino acids in yeast mitochondria.
[0017] The following sequences conform with 37 C.F.R. 1.821-1.825 ("Requirements for Patent Applications Containing Nucleotide Sequences and/or Amino Acid Sequence Disclosures--the Sequence Rules") and are consistent with World Intellectual Property Organization (WIPO) Standard ST.25 (1998) and the sequence listing requirements of the EPO and PCT (Rules 5.2 and 49.5(a-bis), and Section 208 and Annex C of the Administrative Instructions). The symbols and format used for nucleotide and amino acid sequence data comply with the rules set forth in 37 C.F.R. .sctn.1.822.
TABLE-US-00001 TABLE 1 SEQ ID NOs of target proteins and encoding sequences for reduction SEQ ID NO: SEQ ID NO: Organism and gene name nucleic acid amino acid Saccharomyces cerevisiae YJM789, Ilv1 1 2 Schizosaccharomyces pombe, Ilv1 3 4 Candida albicans SC5314, Ilv1 5 6 Candida glabrata, Ilv1 7 8 Kluyveromyces lactis, Ilv1 9 10 Yarrowia lipolytica strain CLIB122, Ilv1 11 12 Pichia stipitis CBS 6054, Ilv1 13 14 Saccharomyces cerevisiae, BAT1 15 16 Schizosaccharomycs pombe, BAT1 17 18 Candida albicans SC5314, BAT1 19 20 Kluyveromyces lactis, BAT1 21 22 Yarrowia lipolytica, BAT1 23 24 Pichia stipitis CBS 6054, BAT1 25 26 Saccharomyces cerevisiae, Leu4 27 28 Schizosaccharomycs pombe, Leu4 29 30 chromosome II Schizosaccharomycs pombe, 31 32 Leu4, NP_596103.2 Candida albicans SC5314, Leu4 33 34 Candida albicans SC5314, Leu4 35 36 Candida albicans SC5314, Leu4 37 38 Candida albicans SC5314, Leu4 39 40 Candida glabrata, Leu4; XP_446653.1 41 42 Candida glabrata, Leu4; XP_446566.1 43 44 Kluyveromyces lactis, Leu4; CAH00792.1 45 46 Kluyveromyces lactis, Leu4; CAG98836.1 47 48 Yarrowia lipolytica, Leu4, CAA88928.1 49 50 Yarrowia lipolytica, Leu4 51 52 Pichia stipitis CBS 6054, Leu4, 53 54 XP_001387341.1 Pichia stipitis CBS 6054, Leu4, 55 56 XP_001384536.2 Saccharomyces cerevisiae, PDB1 57 58 Schizosaccharomycs pombe, PDB1 59 60 Candida albicans SC5314, PDB1 61 62 Kluyveromyces lactis, PDB1 63 64 Yarrowia lipolytica, PDB1 65 66 Pichia stipitis CBS 6054, PDB1 67 68 Saccharomyces cerevisiae, PDA1 69 70 Schizosaccharomycs pombe, PDA1 71 72 Candida albicans, PDA1 73 74 Kluyveromyces lactis, PDA1 75 76 Yarrowia lipolytica, PDA1 hypothetical 77 78 protein Pichia stipitis, PDA1 79 80 Saccharomyces cerevisiae Lat1 pyruvate 105 106 dehydrogenase complex Saccharomyces cerevisiae Lpd1 pyruvate 107 108 dehydrogenase complex Saccharomyces cerevisiae Pdx1 pyruvate 109 110 dehydrogenase complex
TABLE-US-00002 TABLE 2 SEQ ID NOs for primers and vectors Primer or vector name Description SEQ ID NO 112590-88A Primer 81 112590-88B Primer 82 112590-88C Primer 83 112590-88D Primer 84 pUC19-URA3r Vector 85 112590-97A Primer 86 112590-97B Primer 87 112590-49E Primer 88 112590-97C Primer 89 112590-108A Primer 90 112590-108B Primer 91 112590-108C Primer 92 112590-108D Primer 93 112590-108E Primer 94 112590-108F Primer 95 BAT1 check Primer 96 112590-118A Primer 97 112590-118B Primer 98 pRS426::GAL1p-alsS Vector 99 112590-118C Primer 100 112590-118D Primer 101 112590-118E Primer 102 112590-118F Primer 103 112590-118G Primer 104
TABLE-US-00003 TABLE 3 Yeast pyruvate decarboxylase sequences SEQ ID NO: SEQ ID NO: Organism and gene name nucleic acid amino acid PDC1 pyruvate decarboxylase 108 109 from Saccharomyces cerevisiae PDC5 pyruvate decarboxylase 110 111 from Saccharomyces cerevisiae PDC6 pyruvate decarboxylase 112 113 from Saccharomyces cerevisiae Pyruvate decarboxylase 114 115 from Candida glabrata PDC1 pyruvate decarboxylase 116 117 from Pichia stipitis PDC2 pyruvate decarboxylase 118 119 from Pichia stipitis Pyruvate decarboxylase 120 121 from Kluyveromyces lactis Pyruvate decarboxylase 122 123 from Yarrowia lipolytica Pyruvate decarboxylase 124 125 from Schizosaccharomyces pombe
DETAILED DESCRIPTION OF THE INVENTION
[0018] The present invention relates to recombinant yeast cells engineered to have reduced activity of threonine deaminase and isopropylmalate synthase, and optionally reduced activity of branched chain amino acid transaminase, in the mitochondria. These cells produce increased amounts of isobutanol as compared to cells with normal levels of these enzyme activities. Isobutanol is valuable as a fuel or fuel additive to reduce demand for fossil fuels.
[0019] The following abbreviations and definitions will be used for the interpretation of the specification and the claims.
[0020] As used herein, the terms "comprises," "comprising," "includes," "including," "has," "having," "contains" or "containing," or any other variation thereof, are intended to cover a non-exclusive inclusion. For example, a composition, a mixture, process, method, article, or apparatus that comprises a list of elements is not necessarily limited to only those elements but may include other elements not expressly listed or inherent to such composition, mixture, process, method, article, or apparatus. Further, unless expressly stated to the contrary, "or" refers to an inclusive or and not to an exclusive or. For example, a condition A or B is satisfied by any one of the following: A is true (or present) and B is false (or not present), A is false (or not present) and B is true (or present), and both A and B are true (or present).
[0021] Also, the indefinite articles "a" and "an" preceding an element or component of the invention are intended to be nonrestrictive regarding the number of instances (i.e. occurrences) of the element or component. Therefore "a" or "an" should be read to include one or at least one, and the singular word form of the element or component also includes the plural unless the number is obviously meant to be singular.
[0022] The term "invention" or "present invention" as used herein is a non-limiting term and is not intended to refer to any single embodiment of the particular invention but encompasses all possible embodiments as described in the specification and the claims.
[0023] As used herein, the term "about" modifying the quantity of an ingredient or reactant of the invention employed refers to variation in the numerical quantity that can occur, for example, through typical measuring and liquid handling procedures used for making concentrates or use solutions in the real world; through inadvertent error in these procedures; through differences in the manufacture, source, or purity of the ingredients employed to make the compositions or carry out the methods; and the like. The term "about" also encompasses amounts that differ due to different equilibrium conditions for a composition resulting from a particular initial mixture. Whether or not modified by the term "about", the claims include equivalents to the quantities. In one embodiment, the term "about" means within 10% of the reported numerical value, preferably within 5% of the reported numerical value.
[0024] The term "isobutanol biosynthetic pathway" refers to an enzyme pathway to produce isobutanol from pyruvate.
[0025] The term "threonine deaminase refers to an enzyme having the EC number EC 4.3.1.19 (Enzyme Nomenclature 1992, Academic Press, San Diego). Threonine deaminase catalyzes the reaction of threonine to 2-oxobutanoate. This is an enzyme involved in branched chain amino acid biosynthesis, specifically of isoleucine. The gene encoding this enzyme is localized in the yeast nucleus however the encoded protein is transported to the mitochondria where it is metabolically active.
[0026] The term "isopropylmalate synthase" refers to an enzyme having the EC number EC 2.3.3.13 (Enzyme Nomenclature 1992, Academic Press, San Diego). Isopropylmalate synthase catalyzes the reaction of alpha-ketoisovalerate to isopropyl malate. This is an enzyme involved in branched chain amino acid biosynthesis, specifically of leucine. The gene encoding this enzyme is localized in the yeast nucleus however the encoded protein is transported to the mitochondria where it is metabolically active.
[0027] The term "branched chain amino acid transaminase" refers to an enzyme having the EC number EC 2.6.1.42 (Enzyme Nomenclature 1992, Academic Press, San Diego). Branched chain amino acid transaminase catalyzes the reaction of alpha-ketoisovalerate to valine and catalyzes the reaction of 2-keto-3-methyl-valerate to isoleucine. This is an enzyme involved in branched chain amino acid biosynthesis, specifically of valine and isoleucine. The gene encoding this enzyme is localized in the yeast nucleus however the encoded protein is transported to the mitochondria where it is metabolically active.
[0028] The term "pyruvate dehydrogenase" refers to an activity provided by a multienzyme complex that may include proteins PDA1, PDB1, LAT1, LPD1, and PDX1. PDA1 and PDB1 are E1.alpha. and E1.beta. subunits of pyruvate dehydrogenase which has EC number EC 1.2.4.1. LAT1 is dihydrolipoyllysine-residue acetyltransferase, also called dihydrolopoyl transacetylase, which has EC number EC 2.3.1.12. LPD1 is dihydrolipoyl dehydrogenase which has EC number EC 1.8.1.4. Pyruvate dehydrogenase activity catalyzes the reaction of pyruvate to acetyl-CoA. The gene encoding this enzyme is localized in the yeast nucleus however the encoded protein is transported to the mitochondria where it is metabolically active.
[0029] The term "carbon substrate" or "fermentable carbon substrate" refers to a carbon source capable of being metabolized by host organisms of the present invention and particularly carbon sources selected from the group consisting of monosaccharides, oligosaccharides, polysaccharides, and one-carbon substrates or mixtures thereof.
[0030] The term "gene" refers to a nucleic acid fragment that is capable of being expressed as a specific protein, optionally including regulatory sequences preceding (5' non-coding sequences) and following (3' non-coding sequences) the coding sequence. "Native gene" refers to a gene as found in nature with its own regulatory sequences. "Chimeric gene" refers to any gene that is not a native gene, comprising regulatory and coding sequences that are not found together in nature. Accordingly, a chimeric gene may comprise regulatory sequences and coding sequences that are derived from different sources, or regulatory sequences and coding sequences derived from the same source, but arranged in a manner different than that found in nature. "Endogenous gene" refers to a native gene in its natural location in the genome of an organism. A "foreign gene" or "heterologous gene" refers to a gene not normally found in the host organism, but that is introduced into the host organism by gene transfer. "Heterologous gene" includes a native coding region, or portion thereof, that is reintroduced into the source organism in a form that is different from the corresponding native gene. For example, a heterologous gene may include a native coding region that is a portion of a chimeric gene including non-native regulatory regions that is reintroduced into the native host. Also foreign genes can comprise native genes inserted into a non-native organism, or chimeric genes. A "transgene" is a gene that has been introduced into the genome by a transformation procedure.
[0031] As used herein the term "coding region" refers to a DNA sequence that codes for a specific amino acid sequence. "Suitable regulatory sequences" refer to nucleotide sequences located upstream (5' non-coding sequences), within, or downstream (3' non-coding sequences) of a coding sequence, and which influence the transcription, RNA processing or stability, or translation of the associated coding sequence. Regulatory sequences may include promoters, translation leader sequences, introns, polyadenylation recognition sequences, RNA processing site, effector binding site and stem-loop structure.
[0032] The term "promoter" refers to a DNA sequence capable of controlling the expression of a coding sequence or functional RNA. In general, a coding sequence is located 3' to a promoter sequence. Promoters may be derived in their entirety from a native gene, or be composed of different elements derived from different promoters found in nature, or even comprise synthetic DNA segments. It is understood by those skilled in the art that different promoters may direct the expression of a gene in different tissues or cell types, or at different stages of development, or in response to different environmental or physiological conditions. Promoters which cause a gene to be expressed in most cell types at most times are commonly referred to as "constitutive promoters". It is further recognized that since in most cases the exact boundaries of regulatory sequences have not been completely defined, DNA fragments of different lengths may have identical promoter activity.
[0033] The term "operably linked" refers to the association of nucleic acid sequences on a single nucleic acid fragment so that the function of one is affected by the other. For example, a promoter is operably linked with a coding sequence when it is capable of effecting the expression of that coding sequence (i.e., that the coding sequence is under the transcriptional control of the promoter). Coding sequences can be operably linked to regulatory sequences in sense or antisense orientation.
[0034] The term "expression", as used herein, refers to the transcription and stable accumulation of sense (mRNA) or antisense RNA derived from the nucleic acid fragment of the invention. Expression may also refer to translation of mRNA into a polypeptide.
[0035] The term "substantially devoid" when used in reference to the presence of an enzyme activity in a host cell means that the presences of that enzyme is not detectable using conventional assay methods or is detectable at such low levels that the presence of the enzyme at would not be expected to have any effect on metabolic pathways.
[0036] As used herein the term "transformation" refers to the transfer of a nucleic acid fragment into a host organism, resulting in genetically stable inheritance. Host organisms containing the transformed nucleic acid fragments are referred to as "transgenic" or "recombinant" or "transformed" organisms.
[0037] The terms "plasmid" and "vector" as used herein, refer to an extra chromosomal element often carrying genes which are not part of the central metabolism of the cell, and usually in the form of circular double-stranded DNA molecules. Such elements may be autonomously replicating sequences, genome integrating sequences, phage or nucleotide sequences, linear or circular, of a single- or double-stranded DNA or RNA, derived from any source, in which a number of nucleotide sequences have been joined or recombined into a unique construction which is capable of introducing a promoter fragment and DNA sequence for a selected gene product along with appropriate 3' untranslated sequence into a cell.
[0038] As used herein the term "codon degeneracy" refers to the nature in the genetic code permitting variation of the nucleotide sequence without effecting the amino acid sequence of an encoded polypeptide. The skilled artisan is well aware of the "codon-bias" exhibited by a specific host cell in usage of nucleotide codons to specify a given amino acid. Therefore, when synthesizing a gene for improved expression in a host cell, it is desirable to design the gene such that its frequency of codon usage approaches the frequency of preferred codon usage of the host cell.
[0039] As used herein, an "isolated nucleic acid fragment" or "isolated nucleic acid molecule" will be used interchangeably and will mean a polymer of RNA or DNA that is single- or double-stranded, optionally containing synthetic, non-natural or altered nucleotide bases. An isolated nucleic acid fragment in the form of a polymer of DNA may be comprised of one or more segments of cDNA, genomic DNA or synthetic DNA.
[0040] A nucleic acid fragment is "hybridizable" to another nucleic acid fragment, such as a cDNA, genomic DNA, or RNA molecule, when a single-stranded form of the nucleic acid fragment can anneal to the other nucleic acid fragment under the appropriate conditions of temperature and solution ionic strength. Hybridization and washing conditions are well known and exemplified in Sambrook, J., Fritsch, E. F. and Maniatis, T. Molecular Cloning: A Laboratory Manual, 2.sup.nd ed., Cold Spring Harbor Laboratory: Cold Spring Harbor, N.Y. (1989), particularly Chapter 11 and Table 11.1 therein (entirely incorporated herein by reference). The conditions of temperature and ionic strength determine the "stringency" of the hybridization. Stringency conditions can be adjusted to screen for moderately similar fragments (such as homologous sequences from distantly related organisms), to highly similar fragments (such as genes that duplicate functional enzymes from closely related organisms). Post-hybridization washes determine stringency conditions. One set of preferred conditions uses a series of washes starting with 6.times.SSC, 0.5% SDS at room temperature for 15 min, then repeated with 2.times.SSC, 0.5% SDS at 45.degree. C. for 30 min, and then repeated twice with 0.2.times.SSC, 0.5% SDS at 50.degree. C. for 30 min. A more preferred set of stringent conditions uses higher temperatures in which the washes are identical to those above except for the temperature of the final two 30 min washes in 0.2.times.SSC, 0.5% SDS was increased to 60.degree. C. Another preferred set of highly stringent conditions uses two final washes in 0.1.times.SSC, 0.1% SDS at 65.degree. C. An additional set of stringent conditions include hybridization at 0.1.times.SSC, 0.1% SDS, 65.degree. C. and washes with 2.times.SSC, 0.1% SDS followed by 0.1.times.SSC, 0.1% SDS, for example.
[0041] Hybridization requires that the two nucleic acids contain complementary sequences, although depending on the stringency of the hybridization, mismatches between bases are possible. The appropriate stringency for hybridizing nucleic acids depends on the length of the nucleic acids and the degree of complementation, variables well known in the art. The greater the degree of similarity or homology between two nucleotide sequences, the greater the value of Tm for hybrids of nucleic acids having those sequences. The relative stability (corresponding to higher Tm) of nucleic acid hybridizations decreases in the following order: RNA:RNA, DNA:RNA, DNA:DNA. For hybrids of greater than 100 nucleotides in length, equations for calculating Tm have been derived (see Sambrook et al., supra, 9.50-9.51). For hybridizations with shorter nucleic acids, i.e., oligonucleotides, the position of mismatches becomes more important, and the length of the oligonucleotide determines its specificity (see Sambrook et al., supra, 11.7-11.8). In one embodiment the length for a hybridizable nucleic acid is at least about 10 nucleotides. Preferably a minimum length for a hybridizable nucleic acid is at least about 15 nucleotides; more preferably at least about 20 nucleotides; and most preferably the length is at least about 30 nucleotides. Furthermore, the skilled artisan will recognize that the temperature and wash solution salt concentration may be adjusted as necessary according to factors such as length of the probe.
[0042] A "substantial portion" of an amino acid or nucleotide sequence is that portion comprising enough of the amino acid sequence of a polypeptide or the nucleotide sequence of a gene to putatively identify that polypeptide or gene, either by manual evaluation of the sequence by one skilled in the art, or by computer-automated sequence comparison and identification using algorithms such as BLAST (Altschul, S. F., et al., J. Mol. Biol., 215:403-410 (1993)). In general, a sequence of ten or more contiguous amino acids or thirty or more nucleotides is necessary in order to putatively identify a polypeptide or nucleic acid sequence as homologous to a known protein or gene. Moreover, with respect to nucleotide sequences, gene specific oligonucleotide probes comprising 20-30 contiguous nucleotides may be used in sequence-dependent methods of gene identification (e.g., Southern hybridization) and isolation (e.g., in situ hybridization of bacterial colonies or bacteriophage plaques). In addition, short oligonucleotides of 12-15 bases may be used as amplification primers in PCR in order to obtain a particular nucleic acid fragment comprising the primers. Accordingly, a "substantial portion" of a nucleotide sequence comprises enough of the sequence to specifically identify and/or isolate a nucleic acid fragment comprising the sequence. The instant specification teaches the complete amino acid and nucleotide sequence encoding particular fungal proteins. The skilled artisan, having the benefit of the sequences as reported herein, may now use all or a substantial portion of the disclosed sequences for purposes known to those skilled in this art. Accordingly, the instant invention comprises the complete sequences as reported in the accompanying Sequence Listing, as well as substantial portions of those sequences as defined above.
[0043] The term "complementary" is used to describe the relationship between nucleotide bases that are capable of hybridizing to one another. For example, with respect to DNA, adenosine is complementary to thymine and cytosine is complementary to guanine.
[0044] The term "percent identity", as known in the art, is a relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences. In the art, "identity" also means the degree of sequence relatedness between polypeptide or polynucleotide sequences, as the case may be, as determined by the match between strings of such sequences. "Identity" and "similarity" can be readily calculated by known methods, including but not limited to those described in: 1.) Computational Molecular Biology (Lesk, A. M., Ed.) Oxford University: NY (1988); 2.) Biocomputing: Informatics and Genome Projects (Smith, D. W., Ed.) Academic: NY (1993); 3.) Computer Analysis of Sequence Data, Part I (Griffin, A. M., and Griffin, H. G., Eds.) Humania: NJ (1994); 4.) Sequence Analysis in Molecular Biology (von Heinje, G., Ed.) Academic (1987); and 5.) Sequence Analysis Primer (Gribskov, M. and Devereux, J., Eds.) Stockton: NY (1991).
[0045] Preferred methods to determine identity are designed to give the best match between the sequences tested. Methods to determine identity and similarity are codified in publicly available computer programs. Sequence alignments and percent identity calculations may be performed using the MegAlign.TM. program of the LASERGENE bioinformatics computing suite (DNASTAR Inc., Madison, Wis.). Multiple alignment of the sequences is performed using the "Clustal method of alignment" which encompasses several varieties of the algorithm including the "Clustal V method of alignment" corresponding to the alignment method labeled Clustal V (described by Higgins and Sharp, CABIOS. 5:151-153 (1989); Higgins, D. G. et al., Comput. Appl. Biosci., 8:189-191 (1992)) and found in the MegAlign.TM. program of the LASERGENE bioinformatics computing suite (DNASTAR Inc.). For multiple alignments, the default values correspond to GAP PENALTY=10 and GAP LENGTH PENALTY=10. Default parameters for pairwise alignments and calculation of percent identity of protein sequences using the Clustal method are KTUPLE=1, GAP PENALTY=3, WINDOW=5 and DIAGONALS SAVED=5. For nucleic acids these parameters are KTUPLE=2, GAP PENALTY=5, WINDOW=4 and DIAGONALS SAVED=4. After alignment of the sequences using the Clustal V program, it is possible to obtain a "percent identity" by viewing the "sequence distances" table in the same program. Additionally the "Clustal W method of alignment" is available and corresponds to the alignment method labeled Clustal W (described by Higgins and Sharp, CABIOS. 5:151-153 (1989); Higgins, D. G. et al., Comput. Appl. Biosci. 8:189-191(1992)) and found in the MegAlign.TM. v6.1 program of the LASERGENE bioinformatics computing suite (DNASTAR Inc.). Default parameters for multiple alignment (GAP PENALTY=10, GAP LENGTH PENALTY=0.2, Delay Divergen Seqs(%)=30, DNA Transition Weight=0.5, Protein Weight Matrix=Gonnet Series, DNA Weight Matrix=IUB). After alignment of the sequences using the Clustal W program, it is possible to obtain a "percent identity" by viewing the "sequence distances" table in the same program.
[0046] It is well understood by one skilled in the art that many levels of sequence identity are useful in identifying polypeptides, from other species, wherein such polypeptides have the same or similar function or activity. Useful examples of percent identities include, but are not limited to: 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%, or any integer percentage from 55% to 100% may be useful in describing the present invention, such as 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99%. Suitable nucleic acid fragments not only have the above homologies but typically encode a polypeptide having at least 50 amino acids, preferably at least 100 amino acids, more preferably at least 150 amino acids, still more preferably at least 200 amino acids, and most preferably at least 250 amino acids.
[0047] The term "sequence analysis software" refers to any computer algorithm or software program that is useful for the analysis of nucleotide or amino acid sequences. "Sequence analysis software" may be commercially available or independently developed. Typical sequence analysis software will include, but is not limited to: 1.) the GCG suite of programs (Wisconsin Package Version 9.0, Genetics Computer Group (GCG), Madison, Wis.); 2.) BLASTP, BLASTN, BLASTX (Altschul et al., J. Mol. Biol., 215:403-410 (1990)); 3.) DNASTAR (DNASTAR, Inc. Madison, Wis.); 4.) Sequencher (Gene Codes Corporation, Ann Arbor, Mich.); and 5.) the FASTA program incorporating the Smith-Waterman algorithm (W. R. Pearson, Comput. Methods Genome Res., [Proc. Int. Symp.] (1994), Meeting Date 1992, 111-20. Editor(s): Suhai, Sandor. Plenum: New York, N.Y.). Within the context of this application it will be understood that where sequence analysis software is used for analysis, that the results of the analysis will be based on the "default values" of the program referenced, unless otherwise specified. As used herein "default values" will mean any set of values or parameters that originally load with the software when first initialized.
[0048] Standard recombinant DNA and molecular cloning techniques used here are well known in the art and are described by Sambrook, J., Fritsch, E. F. and Maniatis, T., Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1989) (hereinafter "Maniatis"); and by Silhavy, T. J., Bennan, M. L. and Enquist, L. W., Experiments with Gene Fusions, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1984); and by Ausubel, F. M. et al., Current Protocols in Molecular Biology, published by Greene Publishing Assoc. and Wiley-Interscience (1987). Additional methods used here are in Methods in Enzymology, Volume 194, Guide to Yeast Genetics and Molecular and Cell Biology (Part A, 2004, Christine Guthrie and Gerald R. Fink (Eds.), Elsevier Academic Press, San Diego, Calif.).
Mitochondrial Enzyme Engineering for Isobutanol Production
[0049] Disclosed herein are yeast cells with improved isobutanol production and engineering of endogenous enzyme activities in the mitochondria of yeast cells. This engineering may be performed in any type of yeast cell that is amenable to genetic engineering methods and that naturally produces at least a small amount of isobutanol as a by-product of incomplete amino acid metabolism. Suitable yeasts include, but are not limited to, Saccharomyces, Schizosaccharomyces, Hansenula, Candida, Kluyveromyces, Yarrowia and Pichia. Suitable strains include, but are not limited to, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Kluyveromyces lactis, Kluyveromyces thermotolerans, Candida glabrata, Candida albicans, Pichia stipitis and Yarrowia lipolytica.
[0050] Applicants have found that altering branched chain amino acid biosynthesis and pyruvate metabolism in the mitochondria of yeast can affect the amount of isobutanol produced by the yeast cell. Surprisingly, Applicants found that altered cells had increased isobutanol production using endogenous isobutanol biosynthesis. Applicants found that by eliminating threonine deaminase and isopropylmalate synthase activities in the yeast mitochondria, a nine-fold increase in isobutanol production was achieved. With further elimination of branched chain amino acid aminotransferase activity in the mitochondria, over twelve-fold increase in isobutanol production was achieved. Additional reduction of pyruvate dehydrogenase activity in the mitochondria resulted in over thirteen-fold increase in isobutanol production.
[0051] Mitochondrial biosynthetic pathways for branched chain amino acid biosynthesis are shown in the diagram in FIG. 1. The following enzymes are encoded by the genes labeled as steps (arrows) in the pathways in FIG. 1:
[0052] ILV1: threonine deaminase
[0053] ILV2: acetolactate synthase (ALS)
[0054] ILV3: dihydroxy-acid dehydratase (DHAD)
[0055] ILV5: acetohydroxy acid reductoisomerase (KARI)
[0056] BAT1: branched chain amino acid aminotransferase
[0057] BAT2: branched chain amino acid transaminase
[0058] LEU4: isopropylmalate synthase
[0059] PDA1*: refers to the complex including the components:
[0060] PDA1: pyruvate dehydrogenase E1.alpha. subunit
[0061] PDB1: pyruvate dehydrogenase E1 .beta. subunit
[0062] LAT1: dihydrolipoyllysine-residue acetyltransferase
[0063] LPD1: dihydrolipoyl dehydrogenase
[0064] PDX1: protein X Threonine deaminase, ALS, KARI, DHAD and branched chain amino acid aminotransferase enzyme activities in the mitochondria form a biosynthetic pathway from threonine to isoleucine. ALS, KARI, DHAD and branched chain amino acid aminotransferase enzyme activities in the mitochondria form a biosynthetic pathway from pyruvate to valine. ALS, KARI, DHAD and isopropylmalate synthase enzyme activities in the mitochondria form a biosynthetic pathway from pyruvate to isopropyl malate, which moves to the cytoplasm and is converted to leucine.
[0065] While not intending to suggest a mechanism of the effect of down-regulating or eliminating the activity of various mitochondrial enzymes the presence of the cytosolic valine to isobutanol pathway generally in yeast suggests some possible explanations for the effects seen here. For example, eliminating threonine deaminase activity may affect pathway intermediate flow in the pathway from threonine to isoleucine. Eliminating isopropylmalate synthase activity may reduce metabolism of the .alpha.-ketoisovalerate intermediate in the leucine pathway. Eliminating mitochondrial branched chain amino acid aminotransferase activity may reduce metabolism of the alpha-ketoisovalerate intermediate in the valine pathway that is fully within the mitochondrion. Applicants found that the combination of the loss of threonine deaminase and isopropylmalate synthase activities in the mitochondria was very effective in increasing isobutanol production, suggesting that there was substantial increase in transfer of .alpha.-ketoisovalerate to the cytoplasm, and it was converted to isobutanol. In the present cells the conversion to isobutanol relies on endogenous enzyme activities of the yeast cell. Applicants found that the combination of the loss of threonine deaminase, isopropylmalate synthase, and branched chain amino acid aminotransferase activities in the mitochondria further increased isobutanol production, suggesting that there was further increase in transfer of .alpha.-ketoisovalerate to the cytoplasm, and it was converted to isobutanol.
[0066] In the yeast mitochondria pyruvate is also converted to acetyl-CoA through pyruvate dehydrogenase activity (see FIG. 1). Applicants found that eliminating pyruvate dehydrogenase activity in the mitochondria further increased isobutanol production, suggesting that flow of pyruvate to alpha-ketoisovalerate was increased, as well as transfer of alpha-ketoisovalerate to the cytoplasm with conversion to valine and then to isobutanol. The conversion of pyruvate to acetyl-CoA is catalyzed by a multienzyme pyruvate dehydrogenase complex. The pyruvate dehydrogenase enzyme is one enzyme of the multienzyme pyruvate dehydrogenase complex. Pyruvate dehydrogenase (EC 1.2.4.1) itself has alpha and beta subunits: PDA1 and PDB1, respectively, forming the E1.alpha. and E1.beta. subunits, respectively, of the E1 component. The complex includes an E2 core which has dihydrolipoamide acetyltransferase activity (EC 2.3.1.12) and E3 which has dihydrolipoamide dehydrogenase activity (EC1.8.1.4). E2 may be encoded by LAT1 and E3 by LPD1. An additional complex protein is encoded by PDX1, which links Lat1p to Lpd1. Thus the pyruvate dehydrogenase complex may include PDA1, PDB1, Lat1, Lpd1, and Pdx1, or homologous proteins encoded by genes which may have alternative names in various yeasts. The activity of any of these proteins may be reduced to affect the function of the pyruvate dehydrogenase complex, and thereby affect pyruvate dehydrogenase activity, to prepare a strain of one embodiment of the present invention. In the description below when referring to PDA1, it is understood that PDA1 may be substituted by any of PDB1, LAT1, LPD1, or PDX1, any of which may be modified to reduce pyruvate dehydrogenase activity.
[0067] In the present invention, any yeast enzymes providing threonine deaminase, isopropylmalate synthase, branched chain amino acid aminotransferase, or pyruvate dehydrogenase activities in the mitochondria may be targets for engineering to reduce these activities. Preferably the activity is reduced such that there is substantially no detectable activity of the target enzyme. Yeast cells are engineered to reduce enzyme activity typically by modification of the gene encoding the target enzyme. The genes encoding these enzymes are ILV1, LEU4, BAT1, and PDA1 (and miltienzyme complex genes PDB1, LAT1, LPD1, and PDX1), respectively. Any ILV1, LEU4, BAT1, or PDA1 gene of yeast encoding a mitochondrial targeted protein is a target for engineering for reduced expression of the encoded enzyme activity in the present cells. Examples of target coding region sequences and their encoded proteins from different species of yeast cells are given as SEQ ID NOs: 1-80 and 105-110 in Table 1. Other target proteins, or their encoding sequences, having at least about 70%-75%, 75%-80%, 80-85%, 85%-90%, 90%-95%, or at least about 98% sequence identity to any of the proteins or coding sequences listed in Table 1, and these activities, may be identified in the literature and in bioinformatics databases well known to the skilled person.
[0068] There is cytoplasmic isopropylmalate synthase activity encoded by LEU9 and cytoplasmic branched chain amino acid transaminase activity encoded by BAT2, genes which are not targets in the present disclosure.
[0069] Because mitochondrial threonine deaminase, isopropylmalate synthase, branched chain amino acid aminotransferase, and pyruvate dehydrogenase complex enzymes are well known, as well as their encoding genes (ILV1, LEU4, BAT1, PDA1, PDB1, LAT1, LPD1, and PDX1, respectively), one skilled in the art can readily identify these proteins and their encoding genes in yeast cells using bioinformatics approaches, to identify additional target genes for engineering as disclosed herein. Typically BLAST (described above) searching of publicly available databases with known target protein sequences, such as those provided herein, is used to identify homologous proteins and their encoding sequences that may be targeted for inactivation in the present strains. For example, endogenous yeast mitochondrial threonine deaminase proteins having amino acid sequence identities of at least about 70%-75%, 75%-80%, 80-85%, 85%-90%, 90%-95% or 98% sequence identity to any of the threonine deaminase proteins of SEQ ID NOs:2, 4, 6, 8, 10, 12, and 14 may have reduced expression in the present strains. Identities are based on the Clustal W method of alignment using the default parameters of GAP PENALTY=10, GAP LENGTH PENALTY=0.1, and Gonnet 250 series of protein weight matrix.
[0070] In the following description, ILV1 is used as an example, and the same description applies to any of LEU4, BAT1, PDA1, PDB1, LAT1, LPD1, and PDX1 coding regions. The sequences of, for example, the ILV1 coding regions provided herein may be used to identify other homologs in nature. For example each of the threonine deaminase encoding nucleic acid fragments described herein may be used to isolate genes encoding homologous proteins. Isolation of homologous genes using sequence-dependent protocols is well known in the art. Examples of sequence-dependent protocols include, but are not limited to: 1.) methods of nucleic acid hybridization; 2.) methods of DNA and RNA amplification, as exemplified by various uses of nucleic acid amplification technologies [e.g., polymerase chain reaction (PCR), Mullis et al., U.S. Pat. No. 4,683,202; ligase chain reaction (LCR), Tabor, S. et al., Proc. Acad. Sci. USA 82:1074 (1985); or strand displacement amplification (SDA), Walker, et al., Proc. Natl. Acad. Sci. U.S.A., 89:392 (1992)]; and 3.) methods of library construction and screening by complementation.
[0071] For example, genes encoding similar proteins or polypeptides to the threonine deaminase encoding genes provided herein could be isolated directly by using all or a portion of the instant nucleic acid fragments as DNA hybridization probes to screen libraries from any desired organism using methodology well known to those skilled in the art. Specific oligonucleotide probes based upon the disclosed nucleic acid sequences can be designed and synthesized by methods known in the art (Maniatis, supra). Moreover, the entire sequences can be used directly to synthesize DNA probes by methods known to the skilled artisan (e.g., random primers DNA labeling, nick translation or end-labeling techniques), or RNA probes using available in vitro transcription systems. In addition, specific primers can be designed and used to amplify a part of (or full-length of) the instant sequences. The resulting amplification products can be labeled directly during amplification reactions or labeled after amplification reactions, and used as probes to isolate full-length DNA fragments by hybridization under conditions of appropriate stringency.
[0072] Typically, in PCR-type amplification techniques, the primers have different sequences and are not complementary to each other. Depending on the desired test conditions, the sequences of the primers should be designed to provide for both efficient and faithful replication of the target nucleic acid. Methods of PCR primer design are common and well known in the art (Thein and Wallace, "The use of oligonucleotides as specific hybridization probes in the Diagnosis of Genetic Disorders", in Human Genetic Diseases: A Practical Approach, K. E. Davis Ed., (1986) pp 33-50, IRL: Herndon, Va.; and Rychlik, W., In Methods in Molecular Biology, White, B. A. Ed., (1993) Vol. 15, pp 31-39, PCR Protocols: Current Methods and Applications. Humania: Totowa, N.J.).
[0073] Generally two short segments of the described sequences may be used in polymerase chain reaction protocols to amplify longer nucleic acid fragments encoding homologous genes from DNA or RNA. The polymerase chain reaction may also be performed on a library of cloned nucleic acid fragments wherein the sequence of one primer is derived from the described nucleic acid fragments, and the sequence of the other primer takes advantage of the presence of the polyadenylic acid tracts to the 3' end of the mRNA precursor encoding microbial genes.
[0074] Alternatively, the second primer sequence may be based upon sequences derived from the cloning vector. For example, the skilled artisan can follow the RACE protocol (Frohman et al., PNAS USA 85:8998 (1988)) to generate cDNAs by using PCR to amplify copies of the region between a single point in the transcript and the 3' or 5' end. Primers oriented in the 3' and 5' directions can be designed from the instant sequences. Using commercially available 3' RACE or 5' RACE systems (e.g., BRL, Gaithersburg, Md.), specific 3' or 5' cDNA fragments can be isolated (Ohara et al., PNAS USA 86:5673 (1989); Loh et al., Science 243:217 (1989)).
[0075] Alternatively, the provided threonine deaminase encoding sequences may be employed as hybridization reagents for the identification of homologs. The basic components of a nucleic acid hybridization test include a probe, a sample suspected of containing the gene or gene fragment of interest, and a specific hybridization method. Probes are typically single-stranded nucleic acid sequences that are complementary to the nucleic acid sequences to be detected. Probes are "hybridizable" to the nucleic acid sequence to be detected. The probe length can vary from 5 bases to tens of thousands of bases, and will depend upon the specific test to be done. Typically a probe length of about 15 bases to about 30 bases is suitable. Only part of the probe molecule need be complementary to the nucleic acid sequence to be detected. In addition, the complementarity between the probe and the target sequence need not be perfect. Hybridization does occur between imperfectly complementary molecules with the result that a certain fraction of the bases in the hybridized region are not paired with the proper complementary base.
[0076] Hybridization methods are well defined. Typically the probe and sample must be mixed under conditions that will permit nucleic acid hybridization. This involves contacting the probe and sample in the presence of an inorganic or organic salt under the proper concentration and temperature conditions. The probe and sample nucleic acids must be in contact for a long enough time that any possible hybridization between the probe and sample nucleic acid may occur. The concentration of probe or target in the mixture will determine the time necessary for hybridization to occur. The higher the probe or target concentration, the shorter the hybridization incubation time needed. Optionally, a chaotropic agent may be added. The chaotropic agent stabilizes nucleic acids by inhibiting nuclease activity. Furthermore, the chaotropic agent allows sensitive and stringent hybridization of short oligonucleotide probes at room temperature (Van Ness and Chen, Nucl. Acids Res. 19:5143-5151 (1991)). Suitable chaotropic agents include guanidinium chloride, guanidinium thiocyanate, sodium thiocyanate, lithium tetrachloroacetate, sodium perchlorate, rubidium tetrachloroacetate, potassium iodide and cesium trifluoroacetate, among others. Typically, the chaotropic agent will be present at a final concentration of about 3 M. If desired, one can add formamide to the hybridization mixture, typically 30-50% (v/v).
[0077] Various hybridization solutions can be employed. Typically, these comprise from about 20 to 60% volume, preferably 30%, of a polar organic solvent. A common hybridization solution employs about 30-50% v/v formamide, about 0.15 to 1 M sodium chloride, about 0.05 to 0.1 M buffers (e.g., sodium citrate, Tris-HCl, PIPES or HEPES (pH range about 6-9)), about 0.05 to 0.2% detergent (e.g., sodium dodecylsulfate), or between 0.5-20 mM EDTA, FICOLL (Pharmacia Inc.) (about 300-500 kdal), polyvinylpyrrolidone (about 250-500 kdal) and serum albumin. Also included in the typical hybridization solution will be unlabeled carrier nucleic acids from about 0.1 to 5 mg/mL, fragmented nucleic DNA (e.g., calf thymus or salmon sperm DNA, or yeast RNA), and optionally from about 0.5 to 2% wt/vol glycine. Other additives may also be included, such as volume exclusion agents that include a variety of polar water-soluble or swellable agents (e.g., polyethylene glycol), anionic polymers (e.g., polyacrylate or polymethylacrylate) and anionic saccharidic polymers (e.g., dextran sulfate).
[0078] Nucleic acid hybridization is adaptable to a variety of assay formats. One of the most suitable is the sandwich assay format. The sandwich assay is particularly adaptable to hybridization under non-denaturing conditions. A primary component of a sandwich-type assay is a solid support. The solid support has adsorbed to it or covalently coupled to it immobilized nucleic acid probe that is unlabeled and complementary to one portion of the sequence.
[0079] Threonine deaminase and isopropylmalate synthase, and optionally branched chain amino acid aminotransferase and/or pyruvate dehydrogenase activities may be reduced using genetic manipulations that disrupt expression of active enzyme from the target gene. Many methods for genetic modification of target genes are known to one skilled in the art and may be used to create the present yeast strains. Modifications that may be used to reduce or eliminate expression of a target protein are disruptions that include, but are not limited to, deletion of the entire gene or a portion of the gene, inserting a DNA fragment into the gene (in either the promoter or coding region) so that the protein is not expressed or expressed at lower levels, introducing a mutation into the coding region which adds a stop codon or frame shift such that a functional protein is not expressed, and introducing one or more mutations into the coding region to alter amino acids so that a non-functional or a less enzymatically active protein is expressed. In addition, expression of a gene may be blocked by expression of an antisense RNA or an interfering RNA, and constructs may be introduced that result in cosuppression. In addition, the synthesis or stability of the transcript may be lessened by mutation. Similarly the efficiency by which a protein is translated from mRNA may be modulated by mutation. In addition, since the target proteins are all mitochondrial, disruption of mitochondrial localization may be used such as disrupting the mitochondrial targeting signal sequence. All of these methods may be readily practiced by one skilled in the art making use of the known or identified coding sequences as exemplified in Table 1.
[0080] DNA sequences surrounding a target gene coding sequence are also useful in some modification procedures and are available for yeasts such as for Saccharomyces cerevisiae in the complete genome sequence coordinated by Genome Project ID9518 of Genome Projects coordinated by NCBI (National Center for Biotechnology Information) with identifying GOPID #13838. Additional examples of yeast genomic sequences include that of Yarrowia lipolytica, GOPIC #13837, and of Candida albicans, which is included in GPID #10771, #10701 and #16373. Additional genomes have been completely sequenced and annotated and are publicly available for the following yeast strains Candida glabrata CBS 138, Kluyveromyces lactis NRRL Y-1140, Pichia stipitis CBS 6054, and Schizosaccharomyces pombe 972h-.
[0081] In particular, DNA sequences surrounding a target coding sequence are useful for modification methods using homologous recombination. For example, in this method flanking sequences are placed bounding a selectable marker gene to mediate homologous recombination whereby the marker gene replaces the target gene. Also partial target gene sequences and flanking sequences bounding a selectable marker gene may be used to mediate homologous recombination whereby the marker gene replaces a portion of the target gene. In addition, the selectable marker may be bounded by site-specific recombination sites, so that following expression of the corresponding site-specific recombinase, the resistance gene is excised from the target gene without reactivating the latter. The site-specific recombination leaves behind a recombination site which disrupts expression of the target gene encoded protein. The homologous recombination vector may be constructed to also leave a deletion in the target gene following excision of the selectable marker, as is well known to one skilled in the art.
[0082] Deletions may be made using mitotic recombination as described in Wach et al. ((1994) Yeast 10:1793-1808). This method involves preparing a DNA fragment that contains a selectable marker between genomic regions that may be as short as 20 bp, and which bound a target DNA sequence. This DNA fragment can be prepared by PCR amplification of the selectable marker gene using as primers oligonucleotides that hybridize to the ends of the marker gene and that include the genomic regions that can recombine with the yeast genome. The linear DNA fragment can be efficiently transformed into yeast and recombined into the genome resulting in gene replacement including with deletion of the target DNA sequence (as described in Methods in Enzymology, v194, pp 281-301 (1991)).
[0083] Moreover, promoter replacement methods may be used to exchange the endogenous transcriptional control elements allowing another means to modulate expression such as described in Mnaimneh et al. ((2004) Cell 118(1):31-44) and in Example 12 herein.
[0084] In addition, a target gene in any yeast cell may be disrupted using random mutagenesis, which is followed by screening to identify strains with reduced target gene encided activity. Using this type of method, the DNA sequence of any region of the genome affecting expression of a target protein need not be known. Methods for creating genetic mutations are common and well known in the art and may be applied to the exercise of creating rmutants. Commonly used random genetic modification methods (reviewed in Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.) include spontaneous mutagenesis, mutagenesis caused by mutator genes, chemical mutagenesis, irradiation with UV or X-rays, or transposon mutagenesis.
[0085] Chemical mutagenesis of yeast commonly involves treatment of yeast cells with one of the following DNA mutagens: ethyl methanesulfonate (EMS), nitrous acid, diethyl sulfate, or N-methyl-N'-nitro-N-nitroso-guanidine (MNNG). These methods of mutagenesis have been reviewed in Spencer et al (Mutagenesis in Yeast, 1996, Yeast Protocols: Methods in Cell and Molecular Biology. Humana Press, Totowa, N.J.). Chemical mutagenesis with EMS may be performed as described in Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. Irradiation with ultraviolet (UV) light or X-rays can also be used to produce random mutagenesis in yeast cells. The primary effect of mutagenesis by UV irradiation is the formation of pyrimidine dimers which disrupt the fidelity of DNA replication. Protocols for UV-mutagenesis of yeast can be found in Spencer et al (Mutagenesis in Yeast, 1996, Yeast Protocols: Methods in Cell and Molecular Biology. Humana Press, Totowa, N.J.). Introduction of a mutator phenotype can also be used to generate random chromosomal mutations in yeast. Common mutator phenotypes can be obtained through disruption of one or more of the following genes: PMS1, MAGI, RAD18 or RAD51. Restoration of the non-mutator phenotype can be easily obtained by insertion of the wildtype allele. Collections of modified cells produced from any of these or other known random mutagenesis processes may be screened for reduced activity of the target enzyme.
Production of Isobutanol
[0086] Strains of yeast disclosed herein may be grown in fermentation media for production of isobutanol. Suitable carbon substrates may include but are not limited to monosaccharides such as fructose, oligosaccharides such as lactose maltose, galactose, or sucrose, polysaccharides such as starch or cellulose or mixtures thereof and unpurified mixtures from renewable feedstocks such as cheese whey permeate, cornsteep liquor, sugar beet molasses, and barley malt. Other carbon substrates may include ethanol, lactate, succinate, or glycerol.
[0087] Additionally the carbon substrate may also be one-carbon substrates such as carbon dioxide, or methanol for which metabolic conversion into key biochemical intermediates has been demonstrated. In addition to one and two carbon substrates, methylotrophic organisms are also known to utilize a number of other carbon containing compounds such as methylamine, glucosamine and a variety of amino acids for metabolic activity. For example, methylotrophic yeasts are known to utilize the carbon from methylamine to form trehalose or glycerol (Bellion et al., Microb. Growth C1 Compd., [Int. Symp.], 7th (1993), 415-32, Editor(s): Murrell, J. Collin; Kelly, Don P. Publisher: Intercept, Andover, UK). Similarly, various species of Candida will metabolize alanine or oleic acid (Sulter et al., Arch. Microbiol. 153:485-489 (1990)). Hence it is contemplated that the source of carbon utilized in the present invention may encompass a wide variety of carbon containing substrates.
[0088] Although it is contemplated that all of the above mentioned carbon substrates and mixtures thereof are suitable in the present invention, preferred carbon substrates are glucose, fructose, and sucrose, or mixtures of these with C5 sugars such as xylose and/or arabinose for yeasts cells modified to use C5 sugars. Sucrose may be derived from renewable sugar sources such as sugar cane, sugar beets, cassava, sweet sorghum, and mixtures thereof. Glucose and dextrose may be derived from renewable grain sources through saccharification of starch based feedstocks including grains such as corn, wheat, rye, barley, oats, and mixtures thereof. In addition, fermentable sugars may be derived from renewable cellulosic or lignocellulosic biomass through processes of pretreatment and saccharification, as described, for example, in U.S. Patent Application Publication No. 2007/0031918A1, which is herein incorporated by reference. Biomass refers to any cellulosic or lignocellulosic material and includes materials comprising cellulose, and optionally further comprising hemicellulose, lignin, starch, oligosaccharides and/or monosaccharides. Biomass may also comprise additional components, such as protein and/or lipid. Biomass may be derived from a single source, or biomass can comprise a mixture derived from more than one source; for example, biomass may comprise a mixture of corn cobs and corn stover, or a mixture of grass and leaves. Biomass includes, but is not limited to, bioenergy crops, agricultural residues, municipal solid waste, industrial solid waste, sludge from paper manufacture, yard waste, wood and forestry waste. Examples of biomass include, but are not limited to, corn grain, corn cobs, crop residues such as corn husks, corn stover, grasses, wheat, wheat straw, barley, barley straw, hay, rice straw, switchgrass, waste paper, sugar cane bagasse, sorghum, soy, components obtained from milling of grains, trees, branches, roots, leaves, wood chips, sawdust, shrubs and bushes, vegetables, fruits, flowers, animal manure, and mixtures thereof.
[0089] In addition to an appropriate carbon source, fermentation media must contain suitable minerals, salts, cofactors, buffers and other components, known to those skilled in the art, suitable for the growth of the cultures and promotion of the enzymatic pathway necessary for production of the desired product.
Byproduct Formation
[0090] It will be appreciated that reduction and preferably elimination of by-products of carbon metabolism other than carbon dioxide and isobutanol would be advantageous for production of isobutanol. For example microorganisms metabolizing sugar substrates produce a variety of by-products in a mixed acid fermentation (Moat, A. G. et al., MicrobialPhysiology, 4.sub.th edition, John Wiley Publishers, N.Y., 2002). Yeast metabolizing sugar substrates produce a variety of by-products like acids and alcohols such as, but not limited to, formate, lactate, succinate, ethanol, acetate and glycerol. Formation of these byproducts during isobutanol fermentation lower the yield of isobutanol. To prevent yield loss of isobutanol the genes encoding enzyme activities corresponding to byproduct formation can be down-regulated or disrupted using methods described herein and/or known in the art.
[0091] Endogenous pyruvate decarboxylase activity in yeast converts pyruvate to acetaldehyde, which is then converted to ethanol or to acetyl-CoA via acetate. Therefore, endogenous pyruvate decarboxylase activity is a target for reduction of byproduct formation. Yeasts may have one or more genes encoding pyruvate decarboylase. For example, there is one gene encoding pyruvate decarboxylase in Kluyveromyces lactis, while there are three isozymes of pyruvate decarboxylase encoded by the PDC1, PCD5, and PDC6 genes in Saccharomyces cerevisiae, as well as a pyruvate decarboxylase regulatory gene PDC2. Expression of pyruvate decarboxylase from PDC6 is minimal. In yeast strains disclosed herein, the pyruvate decarboxylase activity may be reduced by downregulating or disrupting at least one gene encoding a pyruvate decarboxylase, or a gene regulating pyruvate decarboxylase gene expression as described in U.S. patent application Ser. No. 12/477,942, which is herein incorporated by reference. For example, in S. cerevisiae the PDC1 and PDC5 genes, or all three genes, may be disrupted. Alternatively, pyruvate decarboxylase activity may be reduced by disrupting the PDC2 regulatory gene in S. cerevisiae. In other yeasts, genes encoding pyruvate decarboxylase proteins such as those having at least about 80-85%, 85%-90%, 90%-95%, or at least about 98% sequence identity to PDC1 or PDC5 may be downregulated or disrupted. Examples of yeast pyruvate decarboxylase genes or proteins that may be targeted for downregulation or disruption are listed in Table 3 (SEQ ID NOs: 108, 110, 112, 114, 116, 118, 120, 122, and 124).
[0092] Examples of yeast strains with reduced pyruvate decarboxylase activity due to disruption of pyruvate decarboxylase encoding genes have been reported such as for Saccharomyces in Flikweert et al. (Yeast (1996) 12:247-257), for Kluyveromyces in Bianchi et al. (Mol. Microbiol. (1996) 19(1):27-36), and disruption of the regulatory gene in Hohmann, (Mol Gen Genet. (1993) 241:657-666). Saccharomyces strains having no pyruvate decarboxylase activity are available from the ATCC (Accession #200027 and #200028).
Culture Conditions
[0093] Typically cells are grown at a temperature in the range of about 20.degree. C. to about 37.degree. C. in an appropriate medium. Suitable growth media in the present invention are common commercially prepared media such as broth that includes yeast nitrogen base, ammonium sulfate, and dextrose as the carbon/energy source) or YPD Medium, a blend of peptone, yeast extract, and dextrose in optimal proportions for growing most Saccharomyces cerevisiae strains. Other defined or synthetic growth media may also be used and the appropriate medium for growth of the particular microorganism will be known by one skilled in the art of microbiology or fermentation science.
[0094] Suitable pH ranges for the fermentation are between pH 3.0 to pH 7.5, where pH 4.5.0 to pH 6.5 is preferred as the initial condition.
[0095] Fermentations may be performed under aerobic or anaerobic conditions, where anaerobic or microaerobic conditions are preferred.
[0096] The amount of isobutanol produced in the fermentation medium can be determined using a number of methods known in the art, for example, high performance liquid chromatography (HPLC) or gas chromatography (GC).
Industrial Batch and Continuous Fermentations
[0097] The present process employs a batch method of fermentation. A classical batch fermentation is a closed system where the composition of the medium is set at the beginning of the fermentation and not subject to artificial alterations during the fermentation. Thus, at the beginning of the fermentation the medium is inoculated with the desired organism or organisms, and fermentation is permitted to occur without adding anything to the system. Typically, however, a "batch" fermentation is batch with respect to the addition of carbon source and attempts are often made at controlling factors such as pH and oxygen concentration. In batch systems the metabolite and biomass compositions of the system change constantly up to the time the fermentation is stopped. Within batch cultures cells moderate through a static lag phase to a high growth log phase and finally to a stationary phase where growth rate is diminished or halted. If untreated, cells in the stationary phase will eventually die. Cells in log phase generally are responsible for the bulk of production of end product or intermediate.
[0098] A variation on the standard batch system is the Fed-Batch system. Fed-Batch fermentation processes are also suitable in the present invention and comprise a typical batch system with the exception that the substrate is added in increments as the fermentation progresses. Fed-Batch systems are useful when catabolite repression is apt to inhibit the metabolism of the cells and where it is desirable to have limited amounts of substrate in the media. Measurement of the actual substrate concentration in Fed-Batch systems is difficult and is therefore estimated on the basis of the changes of measurable factors such as pH, dissolved oxygen and the partial pressure of waste gases such as CO.sub.2. Batch and Fed-Batch fermentations are common and well known in the art and examples may be found in Thomas D. Brock in Biotechnology: A Textbook of Industrial Microbiology, Second Edition (1989) Sinauer Associates, Inc., Sunderland, Mass., or Deshpande, Mukund V., Appl. Biochem. Biotechnol., 36:227, (1992), herein incorporated by reference.
[0099] Although the present invention is performed in batch mode it is contemplated that the method would be adaptable to continuous fermentation methods. Continuous fermentation is an open system where a defined fermentation medium is added continuously to a bioreactor and an equal amount of conditioned media is removed simultaneously for processing. Continuous fermentation generally maintains the cultures at a constant high density where cells are primarily in log phase growth.
[0100] Continuous fermentation allows for the modulation of one factor or any number of factors that affect cell growth or end product concentration. For example, one method will maintain a limiting nutrient such as the carbon source or nitrogen level at a fixed rate and allow all other parameters to moderate. In other systems a number of factors affecting growth can be altered continuously while the cell concentration, measured by media turbidity, is kept constant. Continuous systems strive to maintain steady state growth conditions and thus the cell loss due to the medium being drawn off must be balanced against the cell growth rate in the fermentation. Methods of modulating nutrients and growth factors for continuous fermentation processes as well as techniques for maximizing the rate of product formation are well known in the art of industrial microbiology and a variety of methods are detailed by Brock, supra.
[0101] It is contemplated that the present invention may be practiced using either batch, fed-batch or continuous processes and that any known mode of fermentation would be suitable. Additionally, it is contemplated that cells may be immobilized on a substrate as whole cell catalysts and subjected to fermentation conditions for isobutanol production.
Methods for Isobutanol Isolation from the Fermentation Medium
[0102] Bioproduced isobutanol may be isolated from the fermentation medium using methods known in the art. For example, solids may be removed from the fermentation medium by centrifugation, filtration, decantation, or the like. Then, the isobutanol may be isolated from the fermentation medium, which has been treated to remove solids as described above, using methods such as distillation, liquid-liquid extraction, or membrane-based separation. Because isobutanol forms a low boiling point, azeotropic mixture with water, distillation can only be used to separate the mixture up to its azeotropic composition. Distillation may be used in combination with another separation method to obtain separation around the azeotrope. Methods that may be used in combination with distillation to isolate and purify isobutanol include, but are not limited to, decantation, liquid-liquid extraction, adsorption, and membrane-based techniques. Additionally, isobutanol may be isolated using azeotropic distillation using an entrainer (see for example Doherty and Malone, Conceptual Design of Distillation Systems, McGraw Hill, New York, 2001).
[0103] The isobutanol-water mixture forms a heterogeneous azeotrope so that distillation may be used in combination with decantation to isolate and purify the isobutanol. In this method, the isobutanol containing fermentation broth is distilled to near the azeotropic composition. Then, the azeotropic mixture is condensed, and the isobutanol is separated from the fermentation medium by decantation. The decanted aqueous phase may be returned to the first distillation column as reflux. The isobutanol-rich decanted organic phase may be further purified by distillation in a second distillation column.
[0104] The isobutanol may also be isolated from the fermentation medium using liquid-liquid extraction in combination with distillation. In this method, the isobutanol is extracted from the fermentation broth using liquid-liquid extraction with a suitable solvent. The isobutanol-containing organic phase is then distilled to separate the isobutanol from the solvent.
[0105] Distillation in combination with adsorption may also be used to isolate isobutanol from the fermentation medium. In this method, the fermentation broth containing the isobutanol is distilled to near the azeotropic composition and then the remaining water is removed by use of an adsorbent, such as molecular sieves (Aden et al. Lignocellulosic Biomass to Ethanol Process Design and Economics Utilizing Co-Current Dilute Acid Prehydrolysis and Enzymatic Hydrolysis for Corn Stover, Report NREL/TP-510-32438, National Renewable Energy Laboratory, June 2002).
[0106] Additionally, distillation in combination with pervaporation may be used to isolate and purify the isobutanol from the fermentation medium. In this method, the fermentation broth containing the isobutanol is distilled to near the azeotropic composition, and then the remaining water is removed by pervaporation through a hydrophilic membrane (Guo et al., J. Membr. Sci. 245, 199-210 (2004)).
EXAMPLES
[0107] The present invention is further defined in the following Examples. It should be understood that these Examples, while indicating preferred embodiments of the invention, are given by way of illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this invention, and without departing from the spirit and scope thereof, can make various changes and modifications of the invention to adapt it to various uses and conditions.
General Methods
[0108] Standard recombinant DNA and molecular cloning techniques used in the Examples are well known in the art and are described by Sambrook, J., Fritsch, E. F. and Maniatis, T. Molecular Cloning: A Laboratory Manual; Cold Spring Harbor Laboratory Press: Cold Spring Harbor, N.Y. (1989) (Maniatis) and by T. J. Silhavy, M. L. Bennan, and L. W. Enquist, Experiments with Gene Fusions, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y. (1984) and by Ausubel, F. M. et al., Current Protocols in Molecular Biology, pub. by Greene Publishing Assoc. and Wiley-Interscience (1987), and by Methods in Yeast Genetics, 2005, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
[0109] Materials and methods suitable for the maintenance and growth of bacterial cultures are well known in the art. Techniques suitable for use in the following Examples may be found as set out in Manual of Methods for General Bacteriology (Phillipp Gerhardt, R. G. E. Murray, Ralph N. Costilow, Eugene W. Nester, Willis A. Wood, Noel R. Krieg and G. Briggs Phillips, eds), American Society for Microbiology, Washington, D.C. (1994)) or by Thomas D. Brock in Biotechnology: A Textbook of Industrial Microbiology, Second Edition, Sinauer Associates, Inc., Sunderland, Mass. (1989). All reagents, restriction enzymes and materials used for the growth and maintenance of microbial cells were obtained from Aldrich Chemicals (Milwaukee, Wis.), BD Diagnostic Systems (Sparks, Md.), Life Technologies (Rockville, Md.), or Sigma Chemical Company (St. Louis, Mo.) unless otherwise specified. Microbial strains were obtained from The American Type Culture Collection (ATCC), Manassas, Va., unless otherwise noted. The oligonucleotide primers used in the following Examples are given in Table 2. All the oligonucleotide primers were synthesized by Sigma-Genosys (Woodlands, Tex.) or Integrated DNA Technologies (Coralsville, Iowa).
[0110] Synthetic complete medium is described in Amberg, Burke and Strathern, 2005, Methods in Yeast Genetics, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.
GC Method
[0111] The GC method utilized an HP-InnoWax column (30 m.times.0.32 mm ID, 0.25 .mu.m film) from Agilent Technologies (Santa Clara, Calif.). The carrier gas was helium at a flow rate of 1 ml/min measured at 150.degree. C. with constant head pressure; injector split was 1:10 at 200.degree. C.; oven temperature was 45.degree. C. for 1 min, 45.degree. C. to 230.degree. C. at 10.degree. C./min, and 230.degree. C. for 30 sec. FID detection was used at 260.degree. C. with 40 ml/min helium makeup gas. Culture broth samples were filtered through 0.2 .mu.M spin filters before injection. Depending on analytical sensitivity desired, either 0.1 .mu.l or 0.5 .mu.l injection volumes were used. Calibrated standard curves were generated for the following compounds: ethanol, isobutanol, acetoin, meso-2,3-butanediol, and (2S,3S)-2,3-butanediol. Analytical standards were also utilized to identify retention times for isobutryaldehyde, isobutyric acid, and isoamyl alcohol.
[0112] SEQ ID NOs for primers and vectors in the examples below are listed in Table 2.
[0113] The meaning of abbreviations is as follows: "s" means second(s), "min" means minute(s), "h" means hour(s), "psi" means pounds per square inch, "nm" means nanometers, "d" means day(s), ".mu.L" means microliter(s), "mL" means milliliter(s), "L" means liter(s), "mm" means millimeter(s), "nm" means nanometers, "mM" means millimolar, "M" means molar, "mmol" means millimole(s), ".mu.mol" means micromole(s)", "g" means gram(s), ".mu.g" means microgram(s) and "ng" means nanogram(s), "PCR" means polymerase chain reaction, "OD" means optical density, "OD.sub.600" means the optical density measured at a wavelength of 600 nm, "kDa" means kilodaltons, "g" means the gravitation constant, "bp" means base pair(s), "kbp" means kilobase pair(s), "% w/v" means weight/volume percent, % v/v" means volume/volume percent, "wt %" means percent by weight, "HPLC" means high performance liquid chromatography, "GC" means gas chromatography, "FID" means flame ionization detector.
Example 1
Eliminating Expression of Enzymes in Branched Chain Amino Acid Biosynthesis Pathways in S. cerevisiae
[0114] Yeast cells were engineered to eliminate activities of specific enzymes of mitochondrial branched chain amino acid biosynthesis. Three chromosomal disruptions were generated in successive fashion in the following genes: ILV1, encoding threonine deaminase; LEU4, encoding 2-isopropylmalate synthase; and BAT1, encoding branched chain amino acid aminotransferase.
[0115] An ilv1::LEU2 cassette was constructed by PCR amplification of the LEU2 marker from pRS425 (ATCC No. 77106) using Phusion DNA polymerase (New England Biolabs Inc., Beverly, Mass.; catalog no. F-540S) with primers 112590-88A (SEQ ID NO:81) and 112590-88B (SEQ ID NO:82). The ILV1 portion of each primer was derived from the 5' region upstream of the ILV1 promoter and 3'region downstream of the transcriptional terminator, respectively, such that integration of the LEU2 marker results in replacement of the ILV1 coding region. The .about.1.7 kb PCR product was transformed into Saccharomyces cerevisiae strain BY4741 (ATCC #201388) with selection on synthetic complete media lacking leucine and supplemented with 2% glucose at 30.degree. C. Transformants were screened by colony PCR using primers 112590-88C (SEQ ID NO:83) and 112590-88D (SEQ ID NO:84). The resulting identified strain had the genotype: BY4741 ilv1::LEU2.
[0116] A leu4::URA3r disruption cassette was constructed by PCR amplification of the URA3r marker from pUC19-URA3r (SEQ ID NO:85) using Phusion DNA polymerase (New England Biolabs Inc., Beverly, Mass.; catalog no. F-540S) and primers 112590-97A (SEQ ID NO:86) and 112590-97B (SEQ ID NO:87). pUC19-URA3r contains the URA3 marker from pRS426 (ATCC no. 77107) flanked by 75 bp homologous repeat sequences to allow homologous recombination in vivo and removal of the URA3 marker. The LEU4 portion of each primer was derived from the 5' region upstream of the LEU4 promoter and 3'region downstream of the transcriptional terminator, respectively, such that integration of the URA3r marker results in replacement of the LEU4 coding region. The .about.1.5 kb PCR product was transformed into BY4741 ilv1::LEU2 cells with selection on synthetic complete media lacking uracil and supplemented with 2% glucose at 30.degree. C. Transformants were screened by PCR using external primers 112590-49E (SEQ ID NO:88) and 112590-97C (SEQ ID NO:89) to verify integration at the correct site. The URA3r marker was recycled by plating on synthetic complete media supplemented with 2% glucose and 5-FOA at 30.degree. C. following standard protocols. Marker removal was confirmed by patching colonies from the 5-FOA plates onto synthetic complete media lacking uracil to verify the absence of growth. The resulting identified strain had the genotype: BY4741 .DELTA.ilv1 .DELTA.leu4.
[0117] A bat1::URA3r disruption cassette was constructed in several steps. A cassette containing the BAT1 5' region was amplified from BY4741 genomic DNA using Phusion DNA polymerase (New England Biolabs Inc., Beverly, Mass.; catalog no. F-540S) and primers 112590-108A (SEQ ID NO:90) and 112590-108B (SEQ ID NO:91). The cassette containing BAT1 3' sequences was amplified from BY4741 genomic DNA using Phusion DNA polymerase and primers 112590-108C (SEQ ID NO:92) and 112590-108D (SEQ ID NO:93). The URA3r marker was PCR-amplified from pUC19-URA3r using Phusion DNA polymerase and primers 112590-108E (SEQ ID NO:94) and 112590-108F (SEQ ID NO:95). The three PCR products were combined in a SOE PCR reaction (Horton et al. (1989) Gene 77:61-68) and amplified using Phusion DNA polymerase and the end primers 112590-108A (SEQ ID NO:90) and 112590-108D (SEQ ID NO:93), generating the full .about.2.8 kb BAT1::URA3r disruption cassette. The BAT1 portion of each primer was derived from the 5' region upstream of the BAT1 promoter and 3'region downstream of the transcriptional terminator, respectively, such that integration of the URA3r marker results in replacement of the BAT1 coding region. The cassette was transformed into BY4741 .DELTA.ilv1 .DELTA.leu4 with selection on synthetic complete media lacking uracil and supplemented with 2% glucose at 30.degree. C. Transformants were screened by PCR using external primers 112590-49E (SEQ ID NO:88) and "BAT1 check" (SEQ ID NO:96) to verify integration at the correct site. The URA3r marker was recycled by plating on synthetic complete media supplemented with 2% glucose and 5-FOA at 30.degree. C. following standard protocols. Marker removal was confirmed by patching colonies from the 5-FOA plates onto synthetic complete media lacking uracil supplemented with 2% glucose to verify the absence of growth. The resulting identified strain had the genotype: BY4741 .DELTA.ilv1 .DELTA.leu4 .DELTA.bat1.
Example 2
Reduction of Pyruvate Dehydrogenase Activity in S. cerevisiae
[0118] To reduce levels of mitochondrial pyruvate dehydrogenase activity, the native promoter of the PDA1 gene, encoding the E1.alpha. subunit of pyruvate dehydrogenase, was replaced with the inducible GAL1 promoter through homologous recombination. The GAL1 promoter and URA3r marker were joined together by SOE PCR. The URA3r marker was PCR amplified from pUC19-URA3r using Phusion DNA polymerase (New England Biolabs Inc., Beverly, Mass.; catalog no. F-540S) and primers 112590-118A (SEQ ID NO:97) and 112590-118B (SEQ ID NO:98). The GAL1 promoter was PCR-amplified from pRS426::GAL1p-alsS (SEQ ID NO:99) using Phusion DNA polymerase (New England Biolabs Inc., Beverly, Mass.; catalog no. F-540S) and primers 112590-118C (SEQ ID NO:100) and 112590-118D (SEQ ID NO:101). pRS426::GAL1p-alsS contained an F1 origin of replication (nt 4976 to 5432) for maintenance in E. coli and a 2 micron origin (nt 2215 to 3560) for replication in yeast. The vector has an GAL1 promoter (nt 7702 to 8144) and CYC1 terminator (nt 5721 to 5970). In addition, it carries the URA3 marker (nt 4042 to 4845) for selection in yeast and ampicillin resistance marker (nt 1225 to 2082) for selection in E. coli.
[0119] The two products were joined by SOE PCR using Phusion DNA polymerase and primers 112590-118E (SEQ ID NO:102) and 112590-118F (SEQ ID NO:103). The PDA1 portion of each primer was derived from the 5' region upstream of the PDA1 promoter and PDA1 coding sequence, respectively, such that integration of the URA3 marker results in replacement of the native PDA1 promoter with the GAL1 promoter. The PCR product was transformed into BY4741 .DELTA.ilv1 .DELTA.leu4 .DELTA.bat1 with selection on synthetic complete media lacking uracil and supplemented with 2% glucose at 30.degree. C. Transformants were screened by PCR using external primers 112590-49E (SEQ ID NO:88) and 112590-118G (SEQ ID NO:104) to verify integration at the PDA1 locus. The URA3r marker was recycled by plating on synthetic complete media supplemented with 2% glucose and 5-FOA at 30.degree. C. following standard protocols. Marker removal was confirmed by patching colonies from the 5FOA plates onto synthetic complete media lacking uracil supplemented with 2% glucose to verify the absence of growth. The resulting identified strain had the genotype: BY4741 .DELTA.ilv1 .DELTA.leu4 .DELTA.bat1 GAL1p-PDA1.
Example 3
Isobutanol Production in S. cerevisiae Deletion Strains
[0120] The purpose of this example is to demonstrate isobutanol production in S. cerevisiae in which ILV1 (threonine deaminase) and LEU4 (2-isopropylmalate synthase), and optionally BAT1 (branched-chain amino acid aminotransferase) genes are disrupted. An additional strain has a PDA1 (pyruvate dehydrogenase) disruption.
[0121] Strains BY4741, BY4741 .DELTA.ilv1 .DELTA.leu4, BY4741 .DELTA.ilv1 .DELTA.leu4 .DELTA.bat1, and BY4741 .DELTA.ilv1 .DELTA.leu4 .DELTA.bat1 GAL1p-PDA1, each transformed with control pR423 (ATCC #77104) and pRS426 plasmids, were grown in synthetic complete media lacking histidine and uracil, and supplemented with 2% glucose under aerobic conditions (20 ml media in 125 ml flask) and cultured at 30.degree. C. with shaking at 220 rpm. Cultures were inoculated at 0.1 OD.sub.600 and assayed for isobutanol titers at 24 hours post-inoculation. Isobutanol was quantitated by GC-FID on a HP-Innowax column using a standard curve of pure isobutanol. A standard curve of isobutanol ranging from 25 mM to 0.6 mM was used to define the linear relationship between raw peak area and isobutanol concentration. Experimental samples were compared against this standard curve to obtain isobutanol titers given in Table 4.
TABLE-US-00004 TABLE 4 Isobutanol production in control and different deletion strains of S. cerevisiae. Strain Isobutanol (mM) BY4741 pRS423/pRS426 0.07* BY4741 .DELTA.ilv1 .DELTA.leu4 pRS423/ 0.64 pRS426 BY4741 .DELTA.ilv1 .DELTA.leu4 .DELTA.bat1 0.90 pRS423/pRS426 BY4741 .DELTA.ilv1 .DELTA.leu4 .DELTA.bat1 0.95 GAL1p-PDA1 pRS423/pRS426 *Sample taken at 48 hours.
Sequence CWU
1
1
12811731DNASaccharomyces cerevisiae 1atgtcagcta ctctactaaa gcaaccatta
tgtacggttg ttcggcaagg taaacagtcc 60aaagtgtctg gattgaacct tttgagacta
aaggctcatt tgcacagaca acacctgtca 120ccttccttga taaaactaca ctctgaattg
aaattggatg agctgcaaac tgataacacc 180cctgattacg tccgtttagt tttaaggtcc
tctgtatacg atgttattaa tgaatctcca 240atctctcaag gtgtaggttt gtcttcccgt
ctaaacacga atgtcatctt gaaaagagaa 300gatctattgc ctgttttctc tttcaagctt
cgtggtgcct ataacatgat tgccaagttg 360gacgattctc aaagaaacca gggtgttatt
gcctgttcag ctgggaatca tgcccaaggt 420gtggcctttg ctgctaaaca cttgaaaata
cctgctacta tcgttatgcc tgtttgtaca 480ccatctatta agtatcaaaa tgtctcgaga
ttagggtctc aagtcgtcct atatggtaac 540gattttgacg aggctaaggc tgaatgtgcc
aaattggctg aagagcgtgg cttgacgaac 600attcctcctt tcgatcatcc ttatgtcatt
gccggtcaag gtactgtagc tatggaaatc 660ctaagacaag tacgtaccgc taataagatc
ggtgctgtct ttgttcccgt cggcggtggt 720ggtttaattg ctggtattgg tgcttatttg
aaaagggttg ctcctcatat caaaatcatt 780ggtgttgaaa cttacgatgc ggccacttta
cataattcct tgcaacgcaa ccagagaact 840cctttacctg tggtgggtac ttttgccgat
ggtacgtctg tgcgtatgat tggtgaagaa 900acatttagag tcgcccaaca agtggttgat
gaagttgttc ttgttaacac tgacgaaatc 960tgtgctgcag taaaggatat ttttgaagat
actagaagta ttgtagaacc atctggtgcc 1020ctttcagtag ccggtatgaa gaaatacatc
tctaccgtac atccagaaat tgaccacact 1080aaaaacacct atgttcccat cctttctggt
gctaacatga actttgatag attaagattt 1140gtttccgaac gtgctgttct tggtgaagga
aaggaagtct tcatgttagt tactttaccc 1200gacgtccctg gtgcgttcaa gaaaatgcaa
aagatcatcc acccaagatc tgttactgaa 1260ttctcttacc gttacaatga acatcgtcat
gagtcctcta gtgaagtgcc caaggcttac 1320atttacactt ctttcagcgt cgttgacaga
gaaaaggaaa tcaagcaagt tatgcaacag 1380ttgaatgctt taggttttga agctgtggat
atctccgata acgaattggc taaatctcat 1440ggtagatact tggttggtgg tgcttctaag
gttcctaatg aaagaattat ttcatttgaa 1500ttccctgaaa gaccaggtgc cttgactagg
ttccttggag gcctaagcga ttcttggaat 1560cttactttat tccattatag aaaccatggt
gccgatatcg gtaaggtttt agctggtatt 1620tccgttcctc caagggaaaa cttaaccttc
caaaaattct tggaagattt aggctacact 1680tatcatgatg aaactgataa cactgtttat
caaaaattct tgaaatatta a 17312576PRTSaccharomyces cerevisiae
2Met Ser Ala Thr Leu Leu Lys Gln Pro Leu Cys Thr Val Val Arg Gln 1
5 10 15 Gly Lys Gln Ser
Lys Val Ser Gly Leu Asn Leu Leu Arg Leu Lys Ala 20
25 30 His Leu His Arg Gln His Leu Ser Pro
Ser Leu Ile Lys Leu His Ser 35 40
45 Glu Leu Lys Leu Asp Glu Leu Gln Thr Asp Asn Thr Pro Asp
Tyr Val 50 55 60
Arg Leu Val Leu Arg Ser Ser Val Tyr Asp Val Ile Asn Glu Ser Pro 65
70 75 80 Ile Ser Gln Gly Val
Gly Leu Ser Ser Arg Leu Asn Thr Asn Val Ile 85
90 95 Leu Lys Arg Glu Asp Leu Leu Pro Val Phe
Ser Phe Lys Leu Arg Gly 100 105
110 Ala Tyr Asn Met Ile Ala Lys Leu Asp Asp Ser Gln Arg Asn Gln
Gly 115 120 125 Val
Ile Ala Cys Ser Ala Gly Asn His Ala Gln Gly Val Ala Phe Ala 130
135 140 Ala Lys His Leu Lys Ile
Pro Ala Thr Ile Val Met Pro Val Cys Thr 145 150
155 160 Pro Ser Ile Lys Tyr Gln Asn Val Ser Arg Leu
Gly Ser Gln Val Val 165 170
175 Leu Tyr Gly Asn Asp Phe Asp Glu Ala Lys Ala Glu Cys Ala Lys Leu
180 185 190 Ala Glu
Glu Arg Gly Leu Thr Asn Ile Pro Pro Phe Asp His Pro Tyr 195
200 205 Val Ile Ala Gly Gln Gly Thr
Val Ala Met Glu Ile Leu Arg Gln Val 210 215
220 Arg Thr Ala Asn Lys Ile Gly Ala Val Phe Val Pro
Val Gly Gly Gly 225 230 235
240 Gly Leu Ile Ala Gly Ile Gly Ala Tyr Leu Lys Arg Val Ala Pro His
245 250 255 Ile Lys Ile
Ile Gly Val Glu Thr Tyr Asp Ala Ala Thr Leu His Asn 260
265 270 Ser Leu Gln Arg Asn Gln Arg Thr
Pro Leu Pro Val Val Gly Thr Phe 275 280
285 Ala Asp Gly Thr Ser Val Arg Met Ile Gly Glu Glu Thr
Phe Arg Val 290 295 300
Ala Gln Gln Val Val Asp Glu Val Val Leu Val Asn Thr Asp Glu Ile 305
310 315 320 Cys Ala Ala Val
Lys Asp Ile Phe Glu Asp Thr Arg Ser Ile Val Glu 325
330 335 Pro Ser Gly Ala Leu Ser Val Ala Gly
Met Lys Lys Tyr Ile Ser Thr 340 345
350 Val His Pro Glu Ile Asp His Thr Lys Asn Thr Tyr Val Pro
Ile Leu 355 360 365
Ser Gly Ala Asn Met Asn Phe Asp Arg Leu Arg Phe Val Ser Glu Arg 370
375 380 Ala Val Leu Gly Glu
Gly Lys Glu Val Phe Met Leu Val Thr Leu Pro 385 390
395 400 Asp Val Pro Gly Ala Phe Lys Lys Met Gln
Lys Ile Ile His Pro Arg 405 410
415 Ser Val Thr Glu Phe Ser Tyr Arg Tyr Asn Glu His Arg His Glu
Ser 420 425 430 Ser
Ser Glu Val Pro Lys Ala Tyr Ile Tyr Thr Ser Phe Ser Val Val 435
440 445 Asp Arg Glu Lys Glu Ile
Lys Gln Val Met Gln Gln Leu Asn Ala Leu 450 455
460 Gly Phe Glu Ala Val Asp Ile Ser Asp Asn Glu
Leu Ala Lys Ser His 465 470 475
480 Gly Arg Tyr Leu Val Gly Gly Ala Ser Lys Val Pro Asn Glu Arg Ile
485 490 495 Ile Ser
Phe Glu Phe Pro Glu Arg Pro Gly Ala Leu Thr Arg Phe Leu 500
505 510 Gly Gly Leu Ser Asp Ser Trp
Asn Leu Thr Leu Phe His Tyr Arg Asn 515 520
525 His Gly Ala Asp Ile Gly Lys Val Leu Ala Gly Ile
Ser Val Pro Pro 530 535 540
Arg Glu Asn Leu Thr Phe Gln Lys Phe Leu Glu Asp Leu Gly Tyr Thr 545
550 555 560 Tyr His Asp
Glu Thr Asp Asn Thr Val Tyr Gln Lys Phe Leu Lys Tyr 565
570 575 31803DNASchizosaccharomyces
pombe 3atgactggaa cgagttttta cacttcggta ctcagattgg gacgattggc tcaacagggc
60ctaaaattcc aatctgtaaa acatattcgt ccatcatgtt tttcatcttt tggattacaa
120gctaaacgtt ggaactctac tcaacaaaat gatagttcta ttgattgttt agaacctaag
180ctgcaaggaa ttattgaaga caatatttct ccctcgacgg cacaaaaaga aatatcagac
240atcaagttta atattccaaa ggaaatgctt cttccagatg gaactcctga ttatttacgt
300ttgactctca cgtctaacgt gtatgaagtt atcaaggaga ctcctcttac aaagggtgtt
360gtcatttctg aaagtaccgg tgttccagtc tacttaaaac gtgaagatct cactcctgtg
420ttttcattta aaattcgagg ggctcataat aaaatggctt ctcttgataa gcagtcattg
480aaaaatggag tcattgcttg ttccgctggc aatcacgccc agggtgttgc ttactccgct
540aggactcttg gtgtaaaagc taccattgtt atgcctcaga atactcctga aatcaaatgg
600aggaacgtta agagattggg cgctaatgtt ctcttacatg gagctaattt tgacattgct
660aaagcagaat gtgcacgttt ggctaaagag caaaatctcg aagttattca tccctttgac
720gatccttatg taattgctgg acaaggaacc attggacttg aaattcttca tcaaatagat
780cttcgcaagc tggatgctat ttactgcgct gttggcggtg gtggtttaat tgctggaata
840gctacttacg ttaagcgtat tgctccccat gttaaggtca ttggtgtcga gacatttgac
900gctgatgctt taaaaaagtc tttgaaggac aaaaagcggg taacccttaa ggaagttggc
960ttattcgctg atggaactgc tgtgaaactt gttggagagg aaaccttccg tcttgtctcc
1020aagaatattg acgatgtagt tcttgttgac aaagatgaga tttgtgcagc cattaaggat
1080gtttttttgg atacccgttc agtggtcgaa ccatcaggag ctatggctgt tgctggtatg
1140aagcgttatg tcgctaaaca caagcctaaa aatcccaatg ctgctcaggt ttgcatctta
1200agtggtgcca atatggactt tgatcgcctt agatttattg ctgagcgtgc tgatcttggt
1260ttgaacaagg aagtattctt gagtgtcact attcctgagc gccctggttc atttgaagcc
1320ctacacaaca ttattactcc acgtagtatt accgaatttt cttatcgtta cgataatgat
1380gactatgcta acatttacac atcgtttgtg gtaaaggacc gtgcaactga attgcctttg
1440attcttcaac aaatctctga gcaaaatatg gttgcagaag atatcagcga taatgaactt
1500gctaaaactc atgcccgtta tcttattgga ggaaaatcat ctgtttcaaa agagcgtttg
1560taccgattgg atttccctga acgccctgga gctttatgta agtttttgag gagtataaag
1620gaagtttgca gcatttccct tttccattat cgtaattgtg gtggagatat agctagtgtg
1680cttgctggcc ttagagtttt tgatggccaa gtggaaaaac ttcattcagt tttggaagag
1740attggataca actgggtgga cgaaacaaat aatcccgttt acttgcgcta tcttcgtaaa
1800tag
18034600PRTSchizosaccharomyces pombe 4Met Thr Gly Thr Ser Phe Tyr Thr Ser
Val Leu Arg Leu Gly Arg Leu 1 5 10
15 Ala Gln Gln Gly Leu Lys Phe Gln Ser Val Lys His Ile Arg
Pro Ser 20 25 30
Cys Phe Ser Ser Phe Gly Leu Gln Ala Lys Arg Trp Asn Ser Thr Gln
35 40 45 Gln Asn Asp Ser
Ser Ile Asp Cys Leu Glu Pro Lys Leu Gln Gly Ile 50
55 60 Ile Glu Asp Asn Ile Ser Pro Ser
Thr Ala Gln Lys Glu Ile Ser Asp 65 70
75 80 Ile Lys Phe Asn Ile Pro Lys Glu Met Leu Leu Pro
Asp Gly Thr Pro 85 90
95 Asp Tyr Leu Arg Leu Thr Leu Thr Ser Asn Val Tyr Glu Val Ile Lys
100 105 110 Glu Thr Pro
Leu Thr Lys Gly Val Val Ile Ser Glu Ser Thr Gly Val 115
120 125 Pro Val Tyr Leu Lys Arg Glu Asp
Leu Thr Pro Val Phe Ser Phe Lys 130 135
140 Ile Arg Gly Ala His Asn Lys Met Ala Ser Leu Asp Lys
Gln Ser Leu 145 150 155
160 Lys Asn Gly Val Ile Ala Cys Ser Ala Gly Asn His Ala Gln Gly Val
165 170 175 Ala Tyr Ser Ala
Arg Thr Leu Gly Val Lys Ala Thr Ile Val Met Pro 180
185 190 Gln Asn Thr Pro Glu Ile Lys Trp Arg
Asn Val Lys Arg Leu Gly Ala 195 200
205 Asn Val Leu Leu His Gly Ala Asn Phe Asp Ile Ala Lys Ala
Glu Cys 210 215 220
Ala Arg Leu Ala Lys Glu Gln Asn Leu Glu Val Ile His Pro Phe Asp 225
230 235 240 Asp Pro Tyr Val Ile
Ala Gly Gln Gly Thr Ile Gly Leu Glu Ile Leu 245
250 255 His Gln Ile Asp Leu Arg Lys Leu Asp Ala
Ile Tyr Cys Ala Val Gly 260 265
270 Gly Gly Gly Leu Ile Ala Gly Ile Ala Thr Tyr Val Lys Arg Ile
Ala 275 280 285 Pro
His Val Lys Val Ile Gly Val Glu Thr Phe Asp Ala Asp Ala Leu 290
295 300 Lys Lys Ser Leu Lys Asp
Lys Lys Arg Val Thr Leu Lys Glu Val Gly 305 310
315 320 Leu Phe Ala Asp Gly Thr Ala Val Lys Leu Val
Gly Glu Glu Thr Phe 325 330
335 Arg Leu Val Ser Lys Asn Ile Asp Asp Val Val Leu Val Asp Lys Asp
340 345 350 Glu Ile
Cys Ala Ala Ile Lys Asp Val Phe Leu Asp Thr Arg Ser Val 355
360 365 Val Glu Pro Ser Gly Ala Met
Ala Val Ala Gly Met Lys Arg Tyr Val 370 375
380 Ala Lys His Lys Pro Lys Asn Pro Asn Ala Ala Gln
Val Cys Ile Leu 385 390 395
400 Ser Gly Ala Asn Met Asp Phe Asp Arg Leu Arg Phe Ile Ala Glu Arg
405 410 415 Ala Asp Leu
Gly Leu Asn Lys Glu Val Phe Leu Ser Val Thr Ile Pro 420
425 430 Glu Arg Pro Gly Ser Phe Glu Ala
Leu His Asn Ile Ile Thr Pro Arg 435 440
445 Ser Ile Thr Glu Phe Ser Tyr Arg Tyr Asp Asn Asp Asp
Tyr Ala Asn 450 455 460
Ile Tyr Thr Ser Phe Val Val Lys Asp Arg Ala Thr Glu Leu Pro Leu 465
470 475 480 Ile Leu Gln Gln
Ile Ser Glu Gln Asn Met Val Ala Glu Asp Ile Ser 485
490 495 Asp Asn Glu Leu Ala Lys Thr His Ala
Arg Tyr Leu Ile Gly Gly Lys 500 505
510 Ser Ser Val Ser Lys Glu Arg Leu Tyr Arg Leu Asp Phe Pro
Glu Arg 515 520 525
Pro Gly Ala Leu Cys Lys Phe Leu Arg Ser Ile Lys Glu Val Cys Ser 530
535 540 Ile Ser Leu Phe His
Tyr Arg Asn Cys Gly Gly Asp Ile Ala Ser Val 545 550
555 560 Leu Ala Gly Leu Arg Val Phe Asp Gly Gln
Val Glu Lys Leu His Ser 565 570
575 Val Leu Glu Glu Ile Gly Tyr Asn Trp Val Asp Glu Thr Asn Asn
Pro 580 585 590 Val
Tyr Leu Arg Tyr Leu Arg Lys 595 600 5
1710DNACandida albicans 5atgtcaatca ctcgactttc aagtgctaag cttttattaa
gtagcacgtc acgcaaacta 60caggtattaa ggttaaatag tacaacgacc aaacccctta
cccctagaca aaaatggccg 120gaacttttgg actctgattt catagtgaat tctcaaggtg
aaaaacaacc cgattatgtc 180aaattgatat taacttcaag agtgtacgat gttgtggacg
aagccggtac accattaacc 240aatgctatca atttatctca tagatgtggt gccaatatct
atcttaaaag agaggatttg 300ttaccagttt tttcgttcaa gttgagaggt gcatataata
tgattgccca tttgcattca 360aattccccac aacctatatc aggggttatt gcttgttcgg
caggaaacca tgcccaagga 420gttgcatttt cttcgagtaa attaaatatc ccagccacaa
ttgtcatgcc tactccaaca 480ccttctatca agtacaccaa tgtttcaaga ttaggtgccc
aagttgtatt gtatggagac 540gattttgatt cagcaaaaca agagtgtgaa aggttgagca
cagagcaaaa tttaatcaac 600attccacctt ttaaccatcc ttacgttatt gctggtcagg
gtacaattgc tttagagatt 660gctagacaat tgagattaga taaattgaat gccatatttg
ttcctgtagg aggaggtggc 720ttaattgcag gtgtggcagt gtatttgaag catattgccc
ctcacgtcaa aatcatagga 780gtagaaacgt atgatgccga tgcattgaac cagtctttaa
agaatagtcg cctggttact 840ttggaaaaag ttggtttgtt tgcagatggt actgccgtga
aagttcttgg agatgaaacc 900tggagattag caaaagaata tgtagatgaa gttgtgcttg
tcaacactga tgaattgtgt 960gctgctatta aggatatttt tgaagacaca aggctgattg
tcgaaccttc tggagcattg 1020tctgttgctg gattgaaaaa gtacattgaa gaacacccag
agattgacca cagagataag 1080acatatgttc cagttttgtc tggtgctaat atgaattttg
atagattaag gtttgttagt 1140gaaagagcag ttttgggtga aggaaaagaa gtctcattgg
ctgttaccat tcctgagaaa 1200cctggtgagt ttgccagatt gcaaaaagtt atcaatccac
gtgctatcac tgaattttca 1260tacaggtaca acggtgaaga aaacgccgat atatttgtgt
cctttaatgt agtggacaag 1320aaaaaagaaa agtcttcagt tatagcagca atggaaaatt
gtggatttga agttgttgat 1380atttcagaaa acgaattggc aaaatctcat ggacgttatt
tagttggtgg taagtcacaa 1440tctacaaaat cctcaaatga aaaaatctat caatttgaat
tccctgaaaa accaaatgct 1500ttgtttaact ttttacaagc attaaggagc gactggaata
tcagcttgtt taattataga 1560aatcatggac atgatgtcgg aaaaatcttg tgtgcattta
ctcttcctga aggatctgag 1620gaagacttcc aagaattttt aaagaatgtt ggttacactt
ttgttgatga atctgataac 1680atcttttaca aaaaattctt gagaagctaa
17106569PRTCandida albicans 6Met Ser Ile Thr Arg
Leu Ser Ser Ala Lys Leu Leu Leu Ser Ser Thr 1 5
10 15 Ser Arg Lys Leu Gln Val Leu Arg Leu Asn
Ser Thr Thr Thr Lys Pro 20 25
30 Leu Thr Pro Arg Gln Lys Trp Pro Glu Leu Leu Asp Ser Asp Phe
Ile 35 40 45 Val
Asn Ser Gln Gly Glu Lys Gln Pro Asp Tyr Val Lys Leu Ile Leu 50
55 60 Thr Ser Arg Val Tyr Asp
Val Val Asp Glu Ala Gly Thr Pro Leu Thr 65 70
75 80 Asn Ala Ile Asn Leu Ser His Arg Cys Gly Ala
Asn Ile Tyr Leu Lys 85 90
95 Arg Glu Asp Leu Leu Pro Val Phe Ser Phe Lys Leu Arg Gly Ala Tyr
100 105 110 Asn Met
Ile Ala His Leu His Ser Asn Ser Pro Gln Pro Ile Ser Gly 115
120 125 Val Ile Ala Cys Ser Ala Gly
Asn His Ala Gln Gly Val Ala Phe Ser 130 135
140 Ser Ser Lys Leu Asn Ile Pro Ala Thr Ile Val Met
Pro Thr Pro Thr 145 150 155
160 Pro Ser Ile Lys Tyr Thr Asn Val Ser Arg Leu Gly Ala Gln Val Val
165 170 175 Leu Tyr Gly
Asp Asp Phe Asp Ser Ala Lys Gln Glu Cys Glu Arg Leu 180
185 190 Ser Thr Glu Gln Asn Leu Ile Asn
Ile Pro Pro Phe Asn His Pro Tyr 195 200
205 Val Ile Ala Gly Gln Gly Thr Ile Ala Leu Glu Ile Ala
Arg Gln Leu 210 215 220
Arg Leu Asp Lys Leu Asn Ala Ile Phe Val Pro Val Gly Gly Gly Gly 225
230 235 240 Leu Ile Ala Gly
Val Ala Val Tyr Leu Lys His Ile Ala Pro His Val 245
250 255 Lys Ile Ile Gly Val Glu Thr Tyr Asp
Ala Asp Ala Leu Asn Gln Ser 260 265
270 Leu Lys Asn Ser Arg Ser Val Thr Leu Glu Lys Val Gly Leu
Phe Ala 275 280 285
Asp Gly Thr Ala Val Lys Val Leu Gly Asp Glu Thr Trp Arg Leu Ala 290
295 300 Lys Glu Tyr Val Asp
Glu Val Val Leu Val Asn Thr Asp Glu Leu Cys 305 310
315 320 Ala Ala Ile Lys Asp Ile Phe Glu Asp Thr
Arg Ser Ile Val Glu Pro 325 330
335 Ser Gly Ala Leu Ser Val Ala Gly Leu Lys Lys Tyr Ile Glu Glu
His 340 345 350 Pro
Glu Ile Asp His Arg Asp Lys Thr Tyr Val Pro Val Leu Ser Gly 355
360 365 Ala Asn Met Asn Phe Asp
Arg Leu Arg Phe Val Ser Glu Arg Ala Val 370 375
380 Leu Gly Glu Gly Lys Glu Val Ser Leu Ala Val
Thr Ile Pro Glu Lys 385 390 395
400 Pro Gly Glu Phe Ala Arg Leu Gln Lys Val Ile Asn Pro Arg Ala Ile
405 410 415 Thr Glu
Phe Ser Tyr Arg Tyr Asn Gly Glu Glu Asn Ala Asp Ile Phe 420
425 430 Val Ser Phe Asn Val Val Asp
Lys Lys Lys Glu Lys Ser Ser Val Ile 435 440
445 Ala Ala Met Glu Asn Cys Gly Phe Glu Val Val Asp
Ile Ser Glu Asn 450 455 460
Glu Leu Ala Lys Ser His Gly Arg Tyr Leu Val Gly Gly Lys Ser Gln 465
470 475 480 Ser Thr Lys
Ser Ser Asn Glu Lys Ile Tyr Gln Phe Glu Phe Pro Glu 485
490 495 Lys Pro Asn Ala Leu Phe Asn Phe
Leu Gln Ala Leu Arg Ser Asp Trp 500 505
510 Asn Ile Ser Leu Phe Asn Tyr Arg Asn His Gly His Asp
Val Gly Lys 515 520 525
Ile Leu Cys Ala Phe Thr Leu Pro Glu Gly Ser Glu Glu Asp Phe Gln 530
535 540 Glu Phe Leu Lys
Asn Val Gly Tyr Thr Phe Val Asp Glu Ser Asp Asn 545 550
555 560 Ile Phe Tyr Lys Lys Phe Leu Arg Ser
565 71716DNACandida glabrata 7atgataccca
agctgctgtg tggtaacaca ctgttgagtg catctgttac aacaagtagg 60tctgtctatg
gcttatctac cagatatttt actcaggatc tcgcaccttc attggttaaa 120ctgcattccg
agttaaagcc tgatgagctt cttactgata acacaccaga ctatgtgcgt 180ttggtgctaa
gatcatcggt ctatgatgtt ataaaggaat cgccaatctc acatggtgtt 240ggtctatcgt
ctagactaaa cacaaatgtc caactgaaaa gagaagattt actaccagtg 300ttctctttca
agctgcgtgg tgcatacaac atgatagcga agttagacga tactcagaga 360aatcaaggtg
tcatcgcatg ttccgcaggt aatcatgcac aaggtgtagc atatgccgct 420agacatttgg
atattccagc aactattgtc atgcctgtgt ctactccatc tataaaatat 480caaaatgtgt
cgagactggg ttcacaagtt gttctatatg gtaatgattt tgacgaagct 540aaagctgaat
gtactaaact ggcagaagag cgtggtttga ctaacatccc tccatttgat 600catccatatg
tcattgctgg tcaaggaaca gttgcgatgg aaatcttgag acaggtctat 660aactcaaata
agatcggtgc tgtctttgtt ccagttggtg gtggtgggtt gattgccggt 720gttggtgcct
atttgaagag agtcactcca cacatcaaga ttataggtgt ggaaacacat 780gatgcagcaa
ctttacacac atctcttcaa agaaataaaa gaacaaatct agctagcgtt 840ggtacttttg
ctgatggtac ttctgtgcgt attattggtg aagaaacctt tagagttgcc 900agagaagttg
tcgatgaaat tgtattggtc aatactgatg aaatttgtgc tgcggttaag 960gatgtctttg
aggataccag aagtattgtt gaaccatctg gtgctcttgc ggttgctggt 1020atgaagaagt
atattactca acttcatcca gaaatagatc actctaagca aacatatgtc 1080ccaattttgt
caggtgctaa tatgaacttc gatagattaa gatttgtttc tgagcgtgct 1140gtattaggtg
aaggtaagga agtttttatg ctggttacca ttcctgacgt tccaggctct 1200ttcaaaaaaa
tgcagaaggt tattcatcca agagctgtta ctgagttctg ttaccgttat 1260aatgaacatc
gtcatgaatc ttctagtgag gttccaaagg cctatatcta tacatctttc 1320agtgtggtag
accgcgaaaa ggagattaaa caagtaatgc agcaactgaa caccctcggt 1380tttgaagccg
tcgatatttc tgacaatgaa ttagcaaaat cacatggtag atatttagtt 1440ggtggtgcgt
caaaggtccc aaatgaaaga attatttcgt tcgaattccc agaaagacct 1500ggggccttaa
ccagattctt ggcaggttta agcgagtctt ggaatttgac attgttccat 1560tacagaaacc
atggtgctga tattggtaaa gtattggctg gtatttctgt gccacctaga 1620gaaaatttaa
ctttccaaaa attcttggaa gatttaggct ataagtacca agatgaaaca 1680gaaaatatgg
tatatcaaag actactgaaa tattaa
17168571PRTCandida glabrata 8Met Ile Pro Lys Leu Leu Cys Gly Asn Thr Leu
Leu Ser Ala Ser Val 1 5 10
15 Thr Thr Ser Arg Ser Val Tyr Gly Leu Ser Thr Arg Tyr Phe Thr Gln
20 25 30 Asp Leu
Ala Pro Ser Leu Val Lys Leu His Ser Glu Leu Lys Pro Asp 35
40 45 Glu Leu Leu Thr Asp Asn Thr
Pro Asp Tyr Val Arg Leu Val Leu Arg 50 55
60 Ser Ser Val Tyr Asp Val Ile Lys Glu Ser Pro Ile
Ser His Gly Val 65 70 75
80 Gly Leu Ser Ser Arg Leu Asn Thr Asn Val Gln Leu Lys Arg Glu Asp
85 90 95 Leu Leu Pro
Val Phe Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Ile 100
105 110 Ala Lys Leu Asp Asp Thr Gln Arg
Asn Gln Gly Val Ile Ala Cys Ser 115 120
125 Ala Gly Asn His Ala Gln Gly Val Ala Tyr Ala Ala Arg
His Leu Asp 130 135 140
Ile Pro Ala Thr Ile Val Met Pro Val Ser Thr Pro Ser Ile Lys Tyr 145
150 155 160 Gln Asn Val Ser
Arg Leu Gly Ser Gln Val Val Leu Tyr Gly Asn Asp 165
170 175 Phe Asp Glu Ala Lys Ala Glu Cys Thr
Lys Leu Ala Glu Glu Arg Gly 180 185
190 Leu Thr Asn Ile Pro Pro Phe Asp His Pro Tyr Val Ile Ala
Gly Gln 195 200 205
Gly Thr Val Ala Met Glu Ile Leu Arg Gln Val Tyr Asn Ser Asn Lys 210
215 220 Ile Gly Ala Val Phe
Val Pro Val Gly Gly Gly Gly Leu Ile Ala Gly 225 230
235 240 Val Gly Ala Tyr Leu Lys Arg Val Thr Pro
His Ile Lys Ile Ile Gly 245 250
255 Val Glu Thr His Asp Ala Ala Thr Leu His Thr Ser Leu Gln Arg
Asn 260 265 270 Lys
Arg Thr Asn Leu Ala Ser Val Gly Thr Phe Ala Asp Gly Thr Ser 275
280 285 Val Arg Ile Ile Gly Glu
Glu Thr Phe Arg Val Ala Arg Glu Val Val 290 295
300 Asp Glu Ile Val Leu Val Asn Thr Asp Glu Ile
Cys Ala Ala Val Lys 305 310 315
320 Asp Val Phe Glu Asp Thr Arg Ser Ile Val Glu Pro Ser Gly Ala Leu
325 330 335 Ala Val
Ala Gly Met Lys Lys Tyr Ile Thr Gln Leu His Pro Glu Ile 340
345 350 Asp His Ser Lys Gln Thr Tyr
Val Pro Ile Leu Ser Gly Ala Asn Met 355 360
365 Asn Phe Asp Arg Leu Arg Phe Val Ser Glu Arg Ala
Val Leu Gly Glu 370 375 380
Gly Lys Glu Val Phe Met Leu Val Thr Ile Pro Asp Val Pro Gly Ser 385
390 395 400 Phe Lys Lys
Met Gln Lys Val Ile His Pro Arg Ala Val Thr Glu Phe 405
410 415 Cys Tyr Arg Tyr Asn Glu His Arg
His Glu Ser Ser Ser Glu Val Pro 420 425
430 Lys Ala Tyr Ile Tyr Thr Ser Phe Ser Val Val Asp Arg
Glu Lys Glu 435 440 445
Ile Lys Gln Val Met Gln Gln Leu Asn Thr Leu Gly Phe Glu Ala Val 450
455 460 Asp Ile Ser Asp
Asn Glu Leu Ala Lys Ser His Gly Arg Tyr Leu Val 465 470
475 480 Gly Gly Ala Ser Lys Val Pro Asn Glu
Arg Ile Ile Ser Phe Glu Phe 485 490
495 Pro Glu Arg Pro Gly Ala Leu Thr Arg Phe Leu Ala Gly Leu
Ser Glu 500 505 510
Ser Trp Asn Leu Thr Leu Phe His Tyr Arg Asn His Gly Ala Asp Ile
515 520 525 Gly Lys Val Leu
Ala Gly Ile Ser Val Pro Pro Arg Glu Asn Leu Thr 530
535 540 Phe Gln Lys Phe Leu Glu Asp Leu
Gly Tyr Lys Tyr Gln Asp Glu Thr 545 550
555 560 Glu Asn Met Val Tyr Gln Arg Leu Leu Lys Tyr
565 570 91704DNAKluyveromyces lactis
9atgctacaat ctatagtgag aactcctaga gttcttcgtg cttcaaatgc attaaaactt
60tctgttcgct gtgttagcac ggaccagttc tctgataatt tgcagaggat gtactcccat
120ttgaaggctg acgaacgatt ggaagatgga tctccagact acgtgcgttt aattttaagg
180tcttctgttt atgaagtcat tgaagagacc cccatttcac gtgcggtgtc attgtcctct
240agactaaaca ctaacgttaa attgaaaaga gaggatttgt tgccagtgtt ttccttcaag
300ctgcgtggtg cttataacat gattgccaag ctagacgaaa cacagaagaa tgctggtgtt
360attgcgtgct ctgctggtaa tcacgcacaa ggtgttgcct tttcaagtaa tcatatgaac
420attccagcta ccattgtgat gcctgtttca acaccatcaa tcaaatatca aaacgtgtcg
480agattaggtg cccaagtggt tctatacggt gacgacttcg atgaagccaa attggaatgt
540gcgaggttag ctgaagaacg tggtatgaca gatattccac catttgatca tccttacgtt
600atcgctggtc aaggtactat tgccatggag attctaagac aagtacaaaa tgggtctaac
660atcggagcag tgttctgtgc cgtaggtggt ggtggtttga tttcaggtat tggttcatac
720ttgaagagaa tcgcacctca tatcaaggtt attggtgtgg aaacttacga tgccgctacg
780ttagatgttt cattaaagaa cggtaaacgt accccattgc caagtgttgg aacgttcgct
840gatggtacct ctgtgaggtt aatcggtgaa gaaacattcc gtgtttgtca agacgtagtc
900gatgaagtta tcttggtgaa caccgatgaa atctgtgctg ccgttaaaga tgtgtttgag
960gacacaagat caattgtcga accaagtggt gctcttgctg ttgccggttt gaaaaaatat
1020gtctctcaac tacaccctga aatagaccat tctaagaaga catacgttcc aattctttcc
1080ggtgccaaca tgaatttcga ccgtttaaga ttcgtctcag aacgtgctgt attgggtgaa
1140ggtaaagaag tgtttatgtt ggtcaccatt ccagatactc caggttcttt caagaagcta
1200cagaatgtga tccatccaag agctgtcact gaattctcat accgttataa cgagcattgt
1260cacgaaaatg actccgatgt accaaccgct tgtatctaca catcttttaa cgtcgttgac
1320cgtgaaaagg aaatcaagca agtggttcaa caattgcatg ctttaggttt cgaagccgta
1380gatatctctg acaacgaaat ggccaagtct cacggtagat atttggtcgg tggtgcttct
1440aaaattgaaa atgagaaagt cattgcattc gaattcccag agagaccagg tgcactaact
1500aaattcttat caggattgaa cgtttcctgg aacttgactt tattccatta cagaaaccat
1560ggcgctgata ttgggaaaat tttggctggt atcagtgtgc ctccacaaga caacgaaatc
1620ttccaaaagt tcttggacga tctaggatat aaatatcaag atgaaactga caatatggtt
1680taccagaagt ttttgaagta ctag
170410567PRTKluyveromyces lactis 10Met Leu Gln Ser Ile Val Arg Thr Pro
Arg Val Leu Arg Ala Ser Asn 1 5 10
15 Ala Leu Lys Leu Ser Val Arg Cys Val Ser Thr Asp Gln Phe
Ser Asp 20 25 30
Asn Leu Gln Arg Met Tyr Ser His Leu Lys Ala Asp Glu Arg Leu Glu
35 40 45 Asp Gly Ser Pro
Asp Tyr Val Arg Leu Ile Leu Arg Ser Ser Val Tyr 50
55 60 Glu Val Ile Glu Glu Thr Pro Ile
Ser Arg Ala Val Ser Leu Ser Ser 65 70
75 80 Arg Leu Asn Thr Asn Val Lys Leu Lys Arg Glu Asp
Leu Leu Pro Val 85 90
95 Phe Ser Phe Lys Leu Arg Gly Ala Tyr Asn Met Ile Ala Lys Leu Asp
100 105 110 Glu Thr Gln
Lys Asn Ala Gly Val Ile Ala Cys Ser Ala Gly Asn His 115
120 125 Ala Gln Gly Val Ala Phe Ser Ser
Asn His Met Asn Ile Pro Ala Thr 130 135
140 Ile Val Met Pro Val Ser Thr Pro Ser Ile Lys Tyr Gln
Asn Val Ser 145 150 155
160 Arg Leu Gly Ala Gln Val Val Leu Tyr Gly Asp Asp Phe Asp Glu Ala
165 170 175 Lys Leu Glu Cys
Ala Arg Leu Ala Glu Glu Arg Gly Met Thr Asp Ile 180
185 190 Pro Pro Phe Asp His Pro Tyr Val Ile
Ala Gly Gln Gly Thr Ile Ala 195 200
205 Met Glu Ile Leu Arg Gln Val Gln Asn Gly Ser Asn Ile Gly
Ala Val 210 215 220
Phe Cys Ala Val Gly Gly Gly Gly Leu Ile Ser Gly Ile Gly Ser Tyr 225
230 235 240 Leu Lys Arg Ile Ala
Pro His Ile Lys Val Ile Gly Val Glu Thr Tyr 245
250 255 Asp Ala Ala Thr Leu Asp Val Ser Leu Lys
Asn Gly Lys Arg Thr Pro 260 265
270 Leu Pro Ser Val Gly Thr Phe Ala Asp Gly Thr Ser Val Arg Leu
Ile 275 280 285 Gly
Glu Glu Thr Phe Arg Val Cys Gln Asp Val Val Asp Glu Val Ile 290
295 300 Leu Val Asn Thr Asp Glu
Ile Cys Ala Ala Val Lys Asp Val Phe Glu 305 310
315 320 Asp Thr Arg Ser Ile Val Glu Pro Ser Gly Ala
Leu Ala Val Ala Gly 325 330
335 Leu Lys Lys Tyr Val Ser Gln Leu His Pro Glu Ile Asp His Ser Lys
340 345 350 Lys Thr
Tyr Val Pro Ile Leu Ser Gly Ala Asn Met Asn Phe Asp Arg 355
360 365 Leu Arg Phe Val Ser Glu Arg
Ala Val Leu Gly Glu Gly Lys Glu Val 370 375
380 Phe Met Leu Val Thr Ile Pro Asp Thr Pro Gly Ser
Phe Lys Lys Leu 385 390 395
400 Gln Asn Val Ile His Pro Arg Ala Val Thr Glu Phe Ser Tyr Arg Tyr
405 410 415 Asn Glu His
Cys His Glu Asn Asp Ser Asp Val Pro Thr Ala Cys Ile 420
425 430 Tyr Thr Ser Phe Asn Val Val Asp
Arg Glu Lys Glu Ile Lys Gln Val 435 440
445 Val Gln Gln Leu His Ala Leu Gly Phe Glu Ala Val Asp
Ile Ser Asp 450 455 460
Asn Glu Met Ala Lys Ser His Gly Arg Tyr Leu Val Gly Gly Ala Ser 465
470 475 480 Lys Ile Glu Asn
Glu Lys Val Ile Ala Phe Glu Phe Pro Glu Arg Pro 485
490 495 Gly Ala Leu Thr Lys Phe Leu Ser Gly
Leu Asn Val Ser Trp Asn Leu 500 505
510 Thr Leu Phe His Tyr Arg Asn His Gly Ala Asp Ile Gly Lys
Ile Leu 515 520 525
Ala Gly Ile Ser Val Pro Pro Gln Asp Asn Glu Ile Phe Gln Lys Phe 530
535 540 Leu Asp Asp Leu Gly
Tyr Lys Tyr Gln Asp Glu Thr Asp Asn Met Val 545 550
555 560 Tyr Gln Lys Phe Leu Lys Tyr
565 111590DNAYarrowia lipolytica 11atgtccgaac ccgactatct
gaagctcatc ttgaagagcc gcgtctacga cgtgtgcaag 60gaaacacctg tgacatctgc
tcatggtctg agcgagaagc tgggctgcaa agtgctgctc 120aagcgggaag atcttcagcc
ggttttctcg ttcaagctgc gaggagccta caacatgatt 180tcgcagctga gtgacgagga
aaagtggaag ggagtgattg cgtgtagcgc cggtaaccat 240gcccaaggag tcgccttttc
agccaactat ctcaacattc cagcgactat tgtcatgccg 300ttggccactc cttccatcaa
gcacagtaat gtttctagac taggtggcaa ggtggttttg 360cacggagacg attttgattc
ggccaaggcc cactgcaagc agctgtgtga gaaatatgga 420ctcacagata tccctccctt
tgatcacccc cacgtgattg caggccaggg aactattggt 480atggagattc ttcgtcaggc
gtcggacaac ctgaaggccg tgtttatctg tgttggaggc 540ggcggtctga ttgccggagt
aggcgcttac atcaagcgga tccagcccga tgtcaaaatc 600attgccgtgg agacctatga
tgcatgtgct ctgaaacaga gtctcatcaa gggcgaacgg 660gtgactctgc ctgaagtcgg
tctgtttgcc gatggagctg ctgtcaagct gtgtggcgag 720gagactttcc gactctgtcg
caagtacgtt gatggagttg tgcttgtgaa cacggacgag 780atctgcgccg ctatcaaaga
tgtatttgag gccactagat cggtggtgga gcctgctggt 840gctctgtcgg tggctggtct
caagaagtac tgctccgacc cctcggccat ttggtggtca 900cctgagtccg attccgcaaa
ggccaatggt atccccacta acgttgccat ctcagaaacc 960gacgagtatc tgtcaattct
ctctggagcc aacatgaact ttgaccggct tcgattcgtg 1020gccgaacgag ctatgcttgg
agaaggaacc gaagtcttca tggtcgtcac catccccgat 1080attcccggag cgtttgaaaa
gctgcacgag atcattctcc ccagagctgt caccgagttc 1140tcctacagaa agaagtccac
tgctgagaac gaagacgcta acatttttgt gtctttttca 1200gtcaaaaacc gacaagagga
aattgcagac gtgctggaaa agctgcaagc tgccggtatg 1260agcggagtcg acgtttcaga
caacgaactg gcaaagaccc acgctagata tctcgtggga 1320ggccagccag acgtgcctaa
tgagagactg ttccggttcg agttccctga acgacccaac 1380gcgctcaaaa acttcctcgg
aggtgtccag acaaagtgga atatcaccct gttccactac 1440agaaacaacg gcagtgatat
tggaaagatt ctgacagcct tggacgtgcc ggaaagcgac 1500aatgaggcgc tcaaggagtt
tcttgagaag ctcaagtacc cctttgtgga ggagacagac 1560aatgtggtgt acaagcagtt
tatgaagtaa 159012529PRTYarrowia
lipolytica 12Met Ser Glu Pro Asp Tyr Leu Lys Leu Ile Leu Lys Ser Arg Val
Tyr 1 5 10 15 Asp
Val Cys Lys Glu Thr Pro Val Thr Ser Ala His Gly Leu Ser Glu
20 25 30 Lys Leu Gly Cys Lys
Val Leu Leu Lys Arg Glu Asp Leu Gln Pro Val 35
40 45 Phe Ser Phe Lys Leu Arg Gly Ala Tyr
Asn Met Ile Ser Gln Leu Ser 50 55
60 Asp Glu Glu Lys Trp Lys Gly Val Ile Ala Cys Ser Ala
Gly Asn His 65 70 75
80 Ala Gln Gly Val Ala Phe Ser Ala Asn Tyr Leu Asn Ile Pro Ala Thr
85 90 95 Ile Val Met Pro
Leu Ala Thr Pro Ser Ile Lys His Ser Asn Val Ser 100
105 110 Arg Leu Gly Gly Lys Val Val Leu His
Gly Asp Asp Phe Asp Ser Ala 115 120
125 Lys Ala His Cys Lys Gln Leu Cys Glu Lys Tyr Gly Leu Thr
Asp Ile 130 135 140
Pro Pro Phe Asp His Pro His Val Ile Ala Gly Gln Gly Thr Ile Gly 145
150 155 160 Met Glu Ile Leu Arg
Gln Ala Ser Asp Asn Leu Lys Ala Val Phe Ile 165
170 175 Cys Val Gly Gly Gly Gly Leu Ile Ala Gly
Val Gly Ala Tyr Ile Lys 180 185
190 Arg Ile Gln Pro Asp Val Lys Ile Ile Ala Val Glu Thr Tyr Asp
Ala 195 200 205 Cys
Ala Leu Lys Gln Ser Leu Ile Lys Gly Glu Arg Val Thr Leu Pro 210
215 220 Glu Val Gly Leu Phe Ala
Asp Gly Ala Ala Val Lys Leu Cys Gly Glu 225 230
235 240 Glu Thr Phe Arg Leu Cys Arg Lys Tyr Val Asp
Gly Val Val Leu Val 245 250
255 Asn Thr Asp Glu Ile Cys Ala Ala Ile Lys Asp Val Phe Glu Ala Thr
260 265 270 Arg Ser
Val Val Glu Pro Ala Gly Ala Leu Ser Val Ala Gly Leu Lys 275
280 285 Lys Tyr Cys Ser Asp Pro Ser
Ala Ile Trp Trp Ser Pro Glu Ser Asp 290 295
300 Ser Ala Lys Ala Asn Gly Ile Pro Thr Asn Val Ala
Ile Ser Glu Thr 305 310 315
320 Asp Glu Tyr Leu Ser Ile Leu Ser Gly Ala Asn Met Asn Phe Asp Arg
325 330 335 Leu Arg Phe
Val Ala Glu Arg Ala Met Leu Gly Glu Gly Thr Glu Val 340
345 350 Phe Met Val Val Thr Ile Pro Asp
Ile Pro Gly Ala Phe Glu Lys Leu 355 360
365 His Glu Ile Ile Leu Pro Arg Ala Val Thr Glu Phe Ser
Tyr Arg Lys 370 375 380
Lys Ser Thr Ala Glu Asn Glu Asp Ala Asn Ile Phe Val Ser Phe Ser 385
390 395 400 Val Lys Asn Arg
Gln Glu Glu Ile Ala Asp Val Leu Glu Lys Leu Gln 405
410 415 Ala Ala Gly Met Ser Gly Val Asp Val
Ser Asp Asn Glu Leu Ala Lys 420 425
430 Thr His Ala Arg Tyr Leu Val Gly Gly Gln Pro Asp Val Pro
Asn Glu 435 440 445
Arg Leu Phe Arg Phe Glu Phe Pro Glu Arg Pro Asn Ala Leu Lys Asn 450
455 460 Phe Leu Gly Gly Val
Gln Thr Lys Trp Asn Ile Thr Leu Phe His Tyr 465 470
475 480 Arg Asn Asn Gly Ser Asp Ile Gly Lys Ile
Leu Thr Ala Leu Asp Val 485 490
495 Pro Glu Ser Asp Asn Glu Ala Leu Lys Glu Phe Leu Glu Lys Leu
Lys 500 505 510 Tyr
Pro Phe Val Glu Glu Thr Asp Asn Val Val Tyr Lys Gln Phe Met 515
520 525 Lys 131620DNAPichia
stipitis 13atgtttttca gtagatctgg agaagttgaa aaatttccaa accttctcga
cgccgatttc 60aacgaagatg gtgatccaga ctacatcaaa ttgatcttga cttcacgagt
gtatgatgtt 120gtggaaaggg caggaacccc tctcacacat gccatcaatt tgtcccataa
gtgcaattca 180aacatctact tgaagagaga ggatttgctt cctgtattct ctttcaaatt
gcgtggagca 240tataatatga tttcacattt gcattctaac tcaaagatgc cactttcggg
tgtaatagct 300tgttctgctg gtaaccatgc tcaaggtgta gcttactctg ccaacagatt
gaaaattcct 360tccactatag ttatgcctac ggctacacct tctatcaagt ataccaatgt
ttcgagactt 420ggatcgcaag ttgttttgta tggtgacgac tttgactcgg ccaagcaaga
atgtgcccgt 480ttgagttcat tgaacaactt gacggatgtg cctcctttcg accatcccta
tgtcatcgct 540ggccagggta ccatagcatt ggagatcacg agacagttgc gcttggataa
gttgaacgca 600ttgtttgtcc ctgttggtgg tggtggtctt attgctggtg tcgctgtcta
cttgaagaag 660attgctcccc atgtgaagat cattggtgta gaaacaaacg atgctgatgc
cttgtaccag 720tcgctcaagg ctaaaaagct ggtggtactt gaccaagttg gtatgtttgc
tgacggaact 780gctgtcaagg tcttaggtaa agaaacctgg agactctgtg aaaacttagt
agacgaagtc 840gttaaggttt ctactgatga gttgtgtgca gcaatcaagg atatctttga
agacacaaga 900ctgattactg aaccatccgg agccttgtct gtagccggct tgaagaagta
cattgaacaa 960aatccagaca ttgaccacag aaacaagttc tatgtgccca tcttgagtgg
tgccaatatg 1020aacttcgaca gattgagatt cgtcagcgag agagctgttc tcggtgaagg
taaagaagtt 1080tcgttggtgg ttactattcc tgaaaagcct ggtgaattcg ccaagttgca
aagtatcatc 1140aatcctagag ccattacaga attctcgtac aggtgtaatg gtgctgatgc
caacatcttt 1200gtttccttca atgttattga caaaaagaag gaattaaccc caattattga
agacatgaac 1260aacaatgaac atggatacga agtagttgat atctctgaca atgaattagc
caagacccat 1320ggtcgttatt tggtcggcgg taagtcctct gaagaagttg ccaatgaaag
attatacagt 1380ttcgaatttc cagaaaagcc tggagcctta ttcaacttct tacaagcttt
gaaggctgat 1440tggaacatta ctttgtttca ttacagaaat cacgggcacg acatcggcaa
ggttttgtgt 1500ggttttacgc ttcctgaagg aacagatgac gcagatttcc agtccttctt
gaatgaactt 1560ggatacaagt tcaatgttga aaatgacaac gttgtctata agaagttctt
gagaagctga 162014539PRTPichia stipitis 14Met Phe Phe Ser Arg Ser Gly
Glu Val Glu Lys Phe Pro Asn Leu Leu 1 5
10 15 Asp Ala Asp Phe Asn Glu Asp Gly Asp Pro Asp
Tyr Ile Lys Leu Ile 20 25
30 Leu Thr Ser Arg Val Tyr Asp Val Val Glu Arg Ala Gly Thr Pro
Leu 35 40 45 Thr
His Ala Ile Asn Leu Ser His Lys Cys Asn Ser Asn Ile Tyr Leu 50
55 60 Lys Arg Glu Asp Leu Leu
Pro Val Phe Ser Phe Lys Leu Arg Gly Ala 65 70
75 80 Tyr Asn Met Ile Ser His Leu His Ser Asn Ser
Lys Met Pro Leu Ser 85 90
95 Gly Val Ile Ala Cys Ser Ala Gly Asn His Ala Gln Gly Val Ala Tyr
100 105 110 Ser Ala
Asn Arg Leu Lys Ile Pro Ser Thr Ile Val Met Pro Thr Ala 115
120 125 Thr Pro Ser Ile Lys Tyr Thr
Asn Val Ser Arg Leu Gly Ser Gln Val 130 135
140 Val Leu Tyr Gly Asp Asp Phe Asp Ser Ala Lys Gln
Glu Cys Ala Arg 145 150 155
160 Leu Ser Ser Leu Asn Asn Leu Thr Asp Val Pro Pro Phe Asp His Pro
165 170 175 Tyr Val Ile
Ala Gly Gln Gly Thr Ile Ala Leu Glu Ile Thr Arg Gln 180
185 190 Leu Arg Leu Asp Lys Leu Asn Ala
Leu Phe Val Pro Val Gly Gly Gly 195 200
205 Gly Leu Ile Ala Gly Val Ala Val Tyr Leu Lys Lys Ile
Ala Pro His 210 215 220
Val Lys Ile Ile Gly Val Glu Thr Asn Asp Ala Asp Ala Leu Tyr Gln 225
230 235 240 Ser Leu Lys Ala
Lys Lys Ser Val Val Leu Asp Gln Val Gly Met Phe 245
250 255 Ala Asp Gly Thr Ala Val Lys Val Leu
Gly Lys Glu Thr Trp Arg Leu 260 265
270 Cys Glu Asn Leu Val Asp Glu Val Val Lys Val Ser Thr Asp
Glu Leu 275 280 285
Cys Ala Ala Ile Lys Asp Ile Phe Glu Asp Thr Arg Ser Ile Thr Glu 290
295 300 Pro Ser Gly Ala Leu
Ser Val Ala Gly Leu Lys Lys Tyr Ile Glu Gln 305 310
315 320 Asn Pro Asp Ile Asp His Arg Asn Lys Phe
Tyr Val Pro Ile Leu Ser 325 330
335 Gly Ala Asn Met Asn Phe Asp Arg Leu Arg Phe Val Ser Glu Arg
Ala 340 345 350 Val
Leu Gly Glu Gly Lys Glu Val Ser Leu Val Val Thr Ile Pro Glu 355
360 365 Lys Pro Gly Glu Phe Ala
Lys Leu Gln Ser Ile Ile Asn Pro Arg Ala 370 375
380 Ile Thr Glu Phe Ser Tyr Arg Cys Asn Gly Ala
Asp Ala Asn Ile Phe 385 390 395
400 Val Ser Phe Asn Val Ile Asp Lys Lys Lys Glu Leu Thr Pro Ile Ile
405 410 415 Glu Asp
Met Asn Asn Asn Glu His Gly Tyr Glu Val Val Asp Ile Ser 420
425 430 Asp Asn Glu Leu Ala Lys Thr
His Gly Arg Tyr Leu Val Gly Gly Lys 435 440
445 Ser Ser Glu Glu Val Ala Asn Glu Arg Leu Tyr Ser
Phe Glu Phe Pro 450 455 460
Glu Lys Pro Gly Ala Leu Phe Asn Phe Leu Gln Ala Leu Lys Ala Asp 465
470 475 480 Trp Asn Ile
Thr Leu Phe His Tyr Arg Asn His Gly His Asp Ile Gly 485
490 495 Lys Val Leu Cys Gly Phe Thr Leu
Pro Glu Gly Thr Asp Asp Ala Asp 500 505
510 Phe Gln Ser Phe Leu Asn Glu Leu Gly Tyr Lys Phe Asn
Val Glu Asn 515 520 525
Asp Asn Val Val Tyr Lys Lys Phe Leu Arg Ser 530 535
151182DNASaccharomyces cerevisiae 15atgttgcaga gacattcctt
gaagttgggg aaattctcca tcagaacact cgctactggt 60gccccattag atgcatccaa
actaaaaatt actagaaacc caaatccatc caagccaaga 120ccaaatgaag aattagtgtt
cggccagaca ttcaccgatc atatgttgac cattccttgg 180tcagccaaag aagggtgggg
cactccacac atcaagcctt acggtaatct ttctcttgac 240ccatctgctt gtgtattcca
ttatgcattt gaattatttg aaggtttgaa agcctacaga 300actcctcaaa atactatcac
catgttccgt ccggataaga acatggcccg tatgaacaag 360tctgccgcta gaatttgttt
gccaactttc gaatctgaag aattgatcaa acttaccggg 420aaattgatcg aacaagataa
acacttggtt cctcaaggta atggttactc attatacatc 480agaccaacaa tgattggtac
atccaagggt ttaggtgttg gcactccctc cgaggctctt 540ctttatgtta ttacttctcc
agtcggtcct tattataaga ctggtttcaa agccgtacgt 600cttgaagcaa cagactatgc
tacaagagct tggccaggtg gtgttggcga caaaaaattg 660ggtgctaact atgccccatg
catcttacct caactacaag ctgccaaaag agggtaccaa 720caaaatctat ggttgttcgg
cccagaaaag aacatcactg aggttggtac tatgaacgtg 780ttcttcgttt tcctcaacaa
agtcactggc aagaaggaat tggttaccgc tccattagat 840ggtaccattt tagaaggtgt
taccagagac tctgttttaa cattggctcg tgacaaacta 900gatcctcaag aatgggacat
caacgagcgt tattacacta ttactgaagt cgccactaga 960gcaaaacaag gtgaactatt
agaagccttc ggttctggta ctgctgctgt cgtttcacct 1020atcaaggaaa ttggctggaa
caacgaagat attcatgttc cactattgcc tggtgaacaa 1080tgtggtgcat tgaccaagca
agttgctcaa tggattgctg atatccaata cggtagagtc 1140aattatggta actggtcaaa
aactgttgcc gacttgaact aa 118216393PRTSaccharomyces
cerevisiae 16Met Leu Gln Arg His Ser Leu Lys Leu Gly Lys Phe Ser Ile Arg
Thr 1 5 10 15 Leu
Ala Thr Gly Ala Pro Leu Asp Ala Ser Lys Leu Lys Ile Thr Arg
20 25 30 Asn Pro Asn Pro Ser
Lys Pro Arg Pro Asn Glu Glu Leu Val Phe Gly 35
40 45 Gln Thr Phe Thr Asp His Met Leu Thr
Ile Pro Trp Ser Ala Lys Glu 50 55
60 Gly Trp Gly Thr Pro His Ile Lys Pro Tyr Gly Asn Leu
Ser Leu Asp 65 70 75
80 Pro Ser Ala Cys Val Phe His Tyr Ala Phe Glu Leu Phe Glu Gly Leu
85 90 95 Lys Ala Tyr Arg
Thr Pro Gln Asn Thr Ile Thr Met Phe Arg Pro Asp 100
105 110 Lys Asn Met Ala Arg Met Asn Lys Ser
Ala Ala Arg Ile Cys Leu Pro 115 120
125 Thr Phe Glu Ser Glu Glu Leu Ile Lys Leu Thr Gly Lys Leu
Ile Glu 130 135 140
Gln Asp Lys His Leu Val Pro Gln Gly Asn Gly Tyr Ser Leu Tyr Ile 145
150 155 160 Arg Pro Thr Met Ile
Gly Thr Ser Lys Gly Leu Gly Val Gly Thr Pro 165
170 175 Ser Glu Ala Leu Leu Tyr Val Ile Thr Ser
Pro Val Gly Pro Tyr Tyr 180 185
190 Lys Thr Gly Phe Lys Ala Val Arg Leu Glu Ala Thr Asp Tyr Ala
Thr 195 200 205 Arg
Ala Trp Pro Gly Gly Val Gly Asp Lys Lys Leu Gly Ala Asn Tyr 210
215 220 Ala Pro Cys Ile Leu Pro
Gln Leu Gln Ala Ala Lys Arg Gly Tyr Gln 225 230
235 240 Gln Asn Leu Trp Leu Phe Gly Pro Glu Lys Asn
Ile Thr Glu Val Gly 245 250
255 Thr Met Asn Val Phe Phe Val Phe Leu Asn Lys Val Thr Gly Lys Lys
260 265 270 Glu Leu
Val Thr Ala Pro Leu Asp Gly Thr Ile Leu Glu Gly Val Thr 275
280 285 Arg Asp Ser Val Leu Thr Leu
Ala Arg Asp Lys Leu Asp Pro Gln Glu 290 295
300 Trp Asp Ile Asn Glu Arg Tyr Tyr Thr Ile Thr Glu
Val Ala Thr Arg 305 310 315
320 Ala Lys Gln Gly Glu Leu Leu Glu Ala Phe Gly Ser Gly Thr Ala Ala
325 330 335 Val Val Ser
Pro Ile Lys Glu Ile Gly Trp Asn Asn Glu Asp Ile His 340
345 350 Val Pro Leu Leu Pro Gly Glu Gln
Cys Gly Ala Leu Thr Lys Gln Val 355 360
365 Ala Gln Trp Ile Ala Asp Ile Gln Tyr Gly Arg Val Asn
Tyr Gly Asn 370 375 380
Trp Ser Lys Thr Val Ala Asp Leu Asn 385 390
171143DNASchizosaccharomyces pombe 17atggttcaaa ctgctgctct ccatggccca
aagcccatgg atagctccca tataaaagtt 60actaatgtta aggagcttaa acctttgccc
gaatggaaga gtttgaagtt tggtgagaat 120tttactgatc atatgcttat tatgaaatgg
aacagagaaa agggttggag tactcctgag 180atcgttccat ttggtaaact ttgctttcac
cctgcttcct ccgttttcca ttatggtttt 240gagtgctttg aaggcatgaa agctttccgt
gacgaaaagg gtgtcccacg tcttttccgt 300cccatcaaga atgctgagcg tatgctttca
actggtactc gtatatctct tccttccttc 360gaccctgctg agcttgctga aattatcaga
aagttcgtcg ctcacgaaaa ccgttgggtc 420cctgatcagc gtggttactc tttgtacatt
cgtcctactt tcattggtac tgatgaagcc 480ttaggtgtcc accattgtga caacgctatg
ctttatgtta ttgcctctcc cgttggcccc 540tactacagct ctggtttcaa ggccgttaag
ctttgttgct ccgaagaatc cgttcgtgct 600tggcctggcg gtactggtca ctacaagctt
ggtggtaact atgctcctag tgttttgcct 660caaaaagagg ctgccaagaa ggggtatgct
cagattctct ggctttatgg agacgaggac 720tacattactg aggttggtac tatgaactgc
tttactgttt ggattaacaa gaatggcgaa 780aaagaaatca ttactgcccc tcttgacggt
atgatcttac ctggtgtcac tcgtgattct 840attttggaaa tttgccgtga acgtctcgca
cctaaaggct ggaagattac tgagggcaag 900tattccatga aagaggttgc tcaagcttct
aaggaaggtc gccttttgga agtctttgga 960gctggtactg ctgcccttgt ttcccccgtc
aaggctatta actacaaggg tactgagtat 1020gaaattccca tgcctgaggg tcaggaagct
ggtcccatca cttctgaaat cagcaaatgg 1080attttggata tccaatacgg caaggaacct
aacaaccctt ggagcgttcc tgctttgcct 1140taa
114318380PRTSchizosaccharomyces pombe
18Met Val Gln Thr Ala Ala Leu His Gly Pro Lys Pro Met Asp Ser Ser 1
5 10 15 His Ile Lys Val
Thr Asn Val Lys Glu Leu Lys Pro Leu Pro Glu Trp 20
25 30 Lys Ser Leu Lys Phe Gly Glu Asn Phe
Thr Asp His Met Leu Ile Met 35 40
45 Lys Trp Asn Arg Glu Lys Gly Trp Ser Thr Pro Glu Ile Val
Pro Phe 50 55 60
Gly Lys Leu Cys Phe His Pro Ala Ser Ser Val Phe His Tyr Gly Phe 65
70 75 80 Glu Cys Phe Glu Gly
Met Lys Ala Phe Arg Asp Glu Lys Gly Val Pro 85
90 95 Arg Leu Phe Arg Pro Ile Lys Asn Ala Glu
Arg Met Leu Ser Thr Gly 100 105
110 Thr Arg Ile Ser Leu Pro Ser Phe Asp Pro Ala Glu Leu Ala Glu
Ile 115 120 125 Ile
Arg Lys Phe Val Ala His Glu Asn Arg Trp Val Pro Asp Gln Arg 130
135 140 Gly Tyr Ser Leu Tyr Ile
Arg Pro Thr Phe Ile Gly Thr Asp Glu Ala 145 150
155 160 Leu Gly Val His His Cys Asp Asn Ala Met Leu
Tyr Val Ile Ala Ser 165 170
175 Pro Val Gly Pro Tyr Tyr Ser Ser Gly Phe Lys Ala Val Lys Leu Cys
180 185 190 Cys Ser
Glu Glu Ser Val Arg Ala Trp Pro Gly Gly Thr Gly His Tyr 195
200 205 Lys Leu Gly Gly Asn Tyr Ala
Pro Ser Val Leu Pro Gln Lys Glu Ala 210 215
220 Ala Lys Lys Gly Tyr Ala Gln Ile Leu Trp Leu Tyr
Gly Asp Glu Asp 225 230 235
240 Tyr Ile Thr Glu Val Gly Thr Met Asn Cys Phe Thr Val Trp Ile Asn
245 250 255 Lys Asn Gly
Glu Lys Glu Ile Ile Thr Ala Pro Leu Asp Gly Met Ile 260
265 270 Leu Pro Gly Val Thr Arg Asp Ser
Ile Leu Glu Ile Cys Arg Glu Arg 275 280
285 Leu Ala Pro Lys Gly Trp Lys Ile Thr Glu Gly Lys Tyr
Ser Met Lys 290 295 300
Glu Val Ala Gln Ala Ser Lys Glu Gly Arg Leu Leu Glu Val Phe Gly 305
310 315 320 Ala Gly Thr Ala
Ala Leu Val Ser Pro Val Lys Ala Ile Asn Tyr Lys 325
330 335 Gly Thr Glu Tyr Glu Ile Pro Met Pro
Glu Gly Gln Glu Ala Gly Pro 340 345
350 Ile Thr Ser Glu Ile Ser Lys Trp Ile Leu Asp Ile Gln Tyr
Gly Lys 355 360 365
Glu Pro Asn Asn Pro Trp Ser Val Pro Ala Leu Pro 370
375 380 191110DNACandida albicans 19atgtcagctc cattagacgc
cagtaaattg gaaatcacta aaactaccaa accaagtgaa 60ccattaccaa aagaagaatt
ggttttcggt aaatcattca ctgaccatat cttagaagtt 120gaatggactg ctgaaaaagg
atggggtgtt ccaactatta aaccatacca caacttttcc 180cttgatccag ccacctgtgt
tttacattat tcttttgagt tatttgaagg tttaaaggca 240taccgtgata gcaatggtaa
aatcagaact tttagaccag acaaaaatat ggaaagaatg 300aatagatcag ctaaaagagc
tgcattacct acatttgatg gtgaagaatt tatcaaatta 360gttgatcaat ttttgttgat
tgaagaaaga tttgttccaa ctggttacgg atattcactt 420tacttgagac caactttaat
tggtacttca attgggttag gtgtcagtgc accaactaaa 480gcattattat atcttattgc
ttcacctgtt ggtccatatt tcagtggtgg tttcaaacca 540gtgtctttgg aagccacaga
ttacgccgta agagcttggc caaaaggtgt tggttcttat 600aaattgggtg caaactatgt
gtcttgtatt gaaccacaaa tggaagctgc caagagaggt 660cattcccaaa atttgtggtt
atttggtgaa gaaggttata ttactgaagt gggtgctatg 720aatgtttttt ttgcattcaa
gaatgccgat ggcactaaag aattggtgac tccgccattg 780gatggtatga tcttgccagg
tgtcactcgt gattctactt tagaattggc taaaagcaaa 840ttaccaagtg attggactgt
caatgaaaga aaattgacta ttcatgaagt taaagaaaga 900gctgctaaag gtgaattagt
tgaagctttc ggtactggta ccgctgctat tgtttcacca 960attgacaaca ttgaattcca
aggcgaacaa attaaggttc cagtttctgc tggtagttcc 1020ggagaaatag ctttgaagat
caatgattgg ataaaggcta ttcaatatgg tgatgaaagt 1080tttaaaaact ggtctagagt
agcccaatag 111020369PRTCandida
albicans 20Met Ser Ala Pro Leu Asp Ala Ser Lys Leu Glu Ile Thr Lys Thr
Thr 1 5 10 15 Lys
Pro Ser Glu Pro Leu Pro Lys Glu Glu Leu Val Phe Gly Lys Ser
20 25 30 Phe Thr Asp His Ile
Leu Glu Val Glu Trp Thr Ala Glu Lys Gly Trp 35
40 45 Gly Val Pro Thr Ile Lys Pro Tyr His
Asn Phe Ser Leu Asp Pro Ala 50 55
60 Thr Cys Val Leu His Tyr Ser Phe Glu Leu Phe Glu Gly
Leu Lys Ala 65 70 75
80 Tyr Arg Asp Ser Asn Gly Lys Ile Arg Thr Phe Arg Pro Asp Lys Asn
85 90 95 Met Glu Arg Met
Asn Arg Ser Ala Lys Arg Ala Ala Leu Pro Thr Phe 100
105 110 Asp Gly Glu Glu Phe Ile Lys Leu Val
Asp Gln Phe Leu Leu Ile Glu 115 120
125 Glu Arg Phe Val Pro Thr Gly Tyr Gly Tyr Ser Leu Tyr Leu
Arg Pro 130 135 140
Thr Leu Ile Gly Thr Ser Ile Gly Leu Gly Val Ser Ala Pro Thr Lys 145
150 155 160 Ala Leu Leu Tyr Leu
Ile Ala Ser Pro Val Gly Pro Tyr Phe Ser Gly 165
170 175 Gly Phe Lys Pro Val Ser Leu Glu Ala Thr
Asp Tyr Ala Val Arg Ala 180 185
190 Trp Pro Lys Gly Val Gly Ser Tyr Lys Leu Gly Ala Asn Tyr Val
Ser 195 200 205 Cys
Ile Glu Pro Gln Met Glu Ala Ala Lys Arg Gly His Ser Gln Asn 210
215 220 Leu Trp Leu Phe Gly Glu
Glu Gly Tyr Ile Thr Glu Val Gly Ala Met 225 230
235 240 Asn Val Phe Phe Ala Phe Lys Asn Ala Asp Gly
Thr Lys Glu Leu Val 245 250
255 Thr Pro Pro Leu Asp Gly Met Ile Leu Pro Gly Val Thr Arg Asp Ser
260 265 270 Thr Leu
Glu Leu Ala Lys Ser Lys Leu Pro Ser Asp Trp Thr Val Asn 275
280 285 Glu Arg Lys Leu Thr Ile His
Glu Val Lys Glu Arg Ala Ala Lys Gly 290 295
300 Glu Leu Val Glu Ala Phe Gly Thr Gly Thr Ala Ala
Ile Val Ser Pro 305 310 315
320 Ile Asp Asn Ile Glu Phe Gln Gly Glu Gln Ile Lys Val Pro Val Ser
325 330 335 Ala Gly Ser
Ser Gly Glu Ile Ala Leu Lys Ile Asn Asp Trp Ile Lys 340
345 350 Ala Ile Gln Tyr Gly Asp Glu Ser
Phe Lys Asn Trp Ser Arg Val Ala 355 360
365 Gln 211224DNAKluyveromyces lactis 21atgaactaca
ttggactacg gaattgtgca agagctgttt ccagcagagt ttccattcca 60tcaagaggta
ttaagagtca tattttaaca agttatagag ccatgtcctt agacgcatcc 120aaggttaaaa
tcaccaaggt cgaaacccca tcgaagccac gtccaaacga tgagttggtt 180ttcggtcaaa
ctttcactga ccatatgtta accatcgaat ggacagctga aaacggttgg 240ggtgtcccag
agattaaacc atacgggaac ttgtcgttag atccatcctc gtgtgtgttc 300cactatgctt
tcgaattgtt cgaaggtttg aaggcgtaca gaaccccaga caacaagatc 360agcatgttcc
gtgctgataa gaatatggaa cgtatgaaca agtcagcagc cagaatctgt 420ttgccatctt
ttaattcgga tgagttgatc aagttgatcg gtaagttgat cgaacaagac 480aagcatttgg
tgcctcaagg tcaaggttac tccttgtaca tcagacctac aatgattggt 540actactaacg
gattgggtgt tggtactcca gacagagctt tgttgtatgt gatcacatct 600ccagtgggac
catattacaa gactgggttc aaagccgtga gattggaagc tacggattat 660gctactagag
cttggccagg tggtgttggt gacaagaagc ttggtgccaa ctacgcacca 720tgtatcttgc
ctcaattgca agctgctgaa cgtggttacc aacaaaactt gtggttgttc 780ggtccagaaa
agaacatcac tgaagtcggt actatgaacg tcttcttcgt gttcaaggac 840tccaagaccg
gcaagaagga attggttact gctccattgg acggtaccat tttggaaggt 900gtcactagag
actctattct acaattggcc agagaaaact tgaactctga cgagtggatc 960gtctctgaac
gttactacac tatcaccgaa gtggaagaaa gagctgccaa gggcgaattg 1020gtcgaagcgt
tcggttccgg taccgctgct gtcgtgtctc caatcaagga aatcggctgg 1080aacggtcacg
atatccaagt gccattgttg cctggtgaac aatgtggtcc attgaccaag 1140caagtggctg
aatggattgc cgatatccaa tatggcagaa aagaacacaa gggatggtcc 1200cgtatcgttg
ctgacttgaa ctaa
122422407PRTKluyveromyces lactis 22Met Asn Tyr Ile Gly Leu Arg Asn Cys
Ala Arg Ala Val Ser Ser Arg 1 5 10
15 Val Ser Ile Pro Ser Arg Gly Ile Lys Ser His Ile Leu Thr
Ser Tyr 20 25 30
Arg Ala Met Ser Leu Asp Ala Ser Lys Val Lys Ile Thr Lys Val Glu
35 40 45 Thr Pro Ser Lys
Pro Arg Pro Asn Asp Glu Leu Val Phe Gly Gln Thr 50
55 60 Phe Thr Asp His Met Leu Thr Ile
Glu Trp Thr Ala Glu Asn Gly Trp 65 70
75 80 Gly Val Pro Glu Ile Lys Pro Tyr Gly Asn Leu Ser
Leu Asp Pro Ser 85 90
95 Ser Cys Val Phe His Tyr Ala Phe Glu Leu Phe Glu Gly Leu Lys Ala
100 105 110 Tyr Arg Thr
Pro Asp Asn Lys Ile Ser Met Phe Arg Ala Asp Lys Asn 115
120 125 Met Glu Arg Met Asn Lys Ser Ala
Ala Arg Ile Cys Leu Pro Ser Phe 130 135
140 Asn Ser Asp Glu Leu Ile Lys Leu Ile Gly Lys Leu Ile
Glu Gln Asp 145 150 155
160 Lys His Leu Val Pro Gln Gly Gln Gly Tyr Ser Leu Tyr Ile Arg Pro
165 170 175 Thr Met Ile Gly
Thr Thr Asn Gly Leu Gly Val Gly Thr Pro Asp Arg 180
185 190 Ala Leu Leu Tyr Val Ile Thr Ser Pro
Val Gly Pro Tyr Tyr Lys Thr 195 200
205 Gly Phe Lys Ala Val Arg Leu Glu Ala Thr Asp Tyr Ala Thr
Arg Ala 210 215 220
Trp Pro Gly Gly Val Gly Asp Lys Lys Leu Gly Ala Asn Tyr Ala Pro 225
230 235 240 Cys Ile Leu Pro Gln
Leu Gln Ala Ala Glu Arg Gly Tyr Gln Gln Asn 245
250 255 Leu Trp Leu Phe Gly Pro Glu Lys Asn Ile
Thr Glu Val Gly Thr Met 260 265
270 Asn Val Phe Phe Val Phe Lys Asp Ser Lys Thr Gly Lys Lys Glu
Leu 275 280 285 Val
Thr Ala Pro Leu Asp Gly Thr Ile Leu Glu Gly Val Thr Arg Asp 290
295 300 Ser Ile Leu Gln Leu Ala
Arg Glu Asn Leu Asn Ser Asp Glu Trp Ile 305 310
315 320 Val Ser Glu Arg Tyr Tyr Thr Ile Thr Glu Val
Glu Glu Arg Ala Ala 325 330
335 Lys Gly Glu Leu Val Glu Ala Phe Gly Ser Gly Thr Ala Ala Val Val
340 345 350 Ser Pro
Ile Lys Glu Ile Gly Trp Asn Gly His Asp Ile Gln Val Pro 355
360 365 Leu Leu Pro Gly Glu Gln Cys
Gly Pro Leu Thr Lys Gln Val Ala Glu 370 375
380 Trp Ile Ala Asp Ile Gln Tyr Gly Arg Lys Glu His
Lys Gly Trp Ser 385 390 395
400 Arg Ile Val Ala Asp Leu Asn 405
231167DNAYarrowia lipolytica 23atgcttcgaa acaacttgag atcgctttcg
cgggccttca gcacctcctc catgcgtctg 60ggcgccggaa tggacgcctc caagctccag
atcaccaaga ccaagtcccc caaggaaaag 120caggccccca aggatctcat tttcggccat
accttcaccg accacatgct gactgtcgag 180tggactgcca aggacggctg ggctgctccc
cagatcaccc cctacggtcc tcttgagctg 240gatccctccg ccgtcgtcct gcactatgcc
tttgagtgtt tcgagggcct caaggcttac 300aaggacgagt ctggaaacgt gcgtctgttc
cgagtcgaca agaacatgca ccgaatgaac 360acatcggccg agcgaatctg cctgcccgag
tttgatggcg ccgaggctgc caagctgatt 420ggccaattgg ccaagcttga ttccgcttgg
atccccgagg gacgaggcta ctccatgtac 480ctccgacctt ctctgattgg aaccaccgcc
gctctcggcg tcggaacccc cgataaggcg 540ctcttttacg tcattgcatc ccccgtcggc
ccctactacc ctaccggatt caaggccgtc 600aagctggagg ctactgacta cgctgtccga
gcctggcctg gaggagtcgg aaacaagaag 660ctgggagcca actacgctcc ctgtatcaag
cctcagcagc aggccgcttc tcgaggctac 720cagcagaacc tgtggctgtt tggcgacgag
ggcaacatca ccgaggtcgg taccatgaac 780gccttctttg tgtttgagcg aaacggcaag
aaggagcttg tcactgctcc tttggacggt 840actattctcg agggtgtcac tcgagactcc
attctggagc tggctcgaga acgattgcct 900tctgctgact ggatcgtttc cgagcgatac
tgcactatta aagaggtcgc ggaggctgcc 960gagaagggcg agcttgttga ggcctttgga
gctggtactg ccgctgttgt ctcgcctatc 1020aaggagattg gatggggaga gaagactatt
aacattcctc tccagcctgg caaggaggcc 1080ggtaagctga ctgagactgt taatgagtgg
attggagata tccagtacgg taaggatgaa 1140tacaagggat ggtctaaggt ggtctaa
116724388PRTYarrowia lipolytica 24Met
Leu Arg Asn Asn Leu Arg Ser Leu Ser Arg Ala Phe Ser Thr Ser 1
5 10 15 Ser Met Arg Leu Gly Ala
Gly Met Asp Ala Ser Lys Leu Gln Ile Thr 20
25 30 Lys Thr Lys Ser Pro Lys Glu Lys Gln Ala
Pro Lys Asp Leu Ile Phe 35 40
45 Gly His Thr Phe Thr Asp His Met Leu Thr Val Glu Trp Thr
Ala Lys 50 55 60
Asp Gly Trp Ala Ala Pro Gln Ile Thr Pro Tyr Gly Pro Leu Glu Leu 65
70 75 80 Asp Pro Ser Ala Val
Val Leu His Tyr Ala Phe Glu Cys Phe Glu Gly 85
90 95 Leu Lys Ala Tyr Lys Asp Glu Ser Gly Asn
Val Arg Leu Phe Arg Val 100 105
110 Asp Lys Asn Met His Arg Met Asn Thr Ser Ala Glu Arg Ile Cys
Leu 115 120 125 Pro
Glu Phe Asp Gly Ala Glu Ala Ala Lys Leu Ile Gly Gln Leu Ala 130
135 140 Lys Leu Asp Ser Ala Trp
Ile Pro Glu Gly Arg Gly Tyr Ser Met Tyr 145 150
155 160 Leu Arg Pro Ser Leu Ile Gly Thr Thr Ala Ala
Leu Gly Val Gly Thr 165 170
175 Pro Asp Lys Ala Leu Phe Tyr Val Ile Ala Ser Pro Val Gly Pro Tyr
180 185 190 Tyr Pro
Thr Gly Phe Lys Ala Val Lys Leu Glu Ala Thr Asp Tyr Ala 195
200 205 Val Arg Ala Trp Pro Gly Gly
Val Gly Asn Lys Lys Leu Gly Ala Asn 210 215
220 Tyr Ala Pro Cys Ile Lys Pro Gln Gln Gln Ala Ala
Ser Arg Gly Tyr 225 230 235
240 Gln Gln Asn Leu Trp Leu Phe Gly Asp Glu Gly Asn Ile Thr Glu Val
245 250 255 Gly Thr Met
Asn Ala Phe Phe Val Phe Glu Arg Asn Gly Lys Lys Glu 260
265 270 Leu Val Thr Ala Pro Leu Asp Gly
Thr Ile Leu Glu Gly Val Thr Arg 275 280
285 Asp Ser Ile Leu Glu Leu Ala Arg Glu Arg Leu Pro Ser
Ala Asp Trp 290 295 300
Ile Val Ser Glu Arg Tyr Cys Thr Ile Lys Glu Val Ala Glu Ala Ala 305
310 315 320 Glu Lys Gly Glu
Leu Val Glu Ala Phe Gly Ala Gly Thr Ala Ala Val 325
330 335 Val Ser Pro Ile Lys Glu Ile Gly Trp
Gly Glu Lys Thr Ile Asn Ile 340 345
350 Pro Leu Gln Pro Gly Lys Glu Ala Gly Lys Leu Thr Glu Thr
Val Asn 355 360 365
Glu Trp Ile Gly Asp Ile Gln Tyr Gly Lys Asp Glu Tyr Lys Gly Trp 370
375 380 Ser Lys Val Val 385
251113DNAPichia stipitis 25atgtctgctc cattagatgc ctccaagctt
gtgatccaca agaccaccac ccccaaggaa 60aagttgccca acgataagtt ggtcttcggc
aagaccttca ccgaccacat gttggaaatc 120gaatggactg ctcaagccgg ctggggcact
cctaccattt ctccctacca caagttgtct 180ttggatcctt ctactgtcgt attgcactac
gcttttgagt tatttgaagg tatgaaagct 240tacagagaca ctgataacaa catcagaacc
ttcagaggtg acaagaacat ggacagaatg 300aacaagtccg ctgacagaat cgccttacca
acctttgatg gtgaagagtt gatgaagctc 360attgatcagt tcttgctcgt agacgaaagc
tttgttccac aaggtgctgg ctactccctt 420tatttaagac caactatgat cggaaccacc
gagtcattgg gtgtaggtac gccagataag 480gcactcttgt atgttattgc atctcccgtt
ggcccttact atggtactgg cttcaagcct 540gtttccttag aagccactga ctatgctgtt
agagcctggc caggtggtgt aggtaacaga 600aagttgggtg ccaactatgc tccttgtgtc
agacctcagt tagaggctgc taagagaggt 660taccaacaaa acttgtggtt attcggagag
gaaggctaca ttaccgaagt cggtaccatg 720aacgctttct ttgtattcaa gaacgctgac
ggcaccaagg agttggccac tgctcctttg 780gatggtacca tcttggaagg tgtcaccaga
gactcgatct tggaactcac cagagaaaga 840ttgccaaaga acgaatgggt agtgtccgaa
cgtaagttca ccattggtga agttgaagaa 900agagctgcca agggtgagtt gatcgaagca
tttggtgctg gtactgctgc tgttgtttct 960cctatcaagt ctattggctg gaagggcaag
gaaatcgaag ttcctttggc tgctggcgat 1020tccggcgaat tgaccgctca agttgctgag
tggatcagaa agatccaata cggtgaagaa 1080cagtacaaaa actggtccag agttgctcaa
tag 111326370PRTPichia stipitis 26Met Ser
Ala Pro Leu Asp Ala Ser Lys Leu Val Ile His Lys Thr Thr 1 5
10 15 Thr Pro Lys Glu Lys Leu Pro
Asn Asp Lys Leu Val Phe Gly Lys Thr 20 25
30 Phe Thr Asp His Met Leu Glu Ile Glu Trp Thr Ala
Gln Ala Gly Trp 35 40 45
Gly Thr Pro Thr Ile Ser Pro Tyr His Lys Leu Ser Leu Asp Pro Ser
50 55 60 Thr Val Val
Leu His Tyr Ala Phe Glu Leu Phe Glu Gly Met Lys Ala 65
70 75 80 Tyr Arg Asp Thr Asp Asn Asn
Ile Arg Thr Phe Arg Gly Asp Lys Asn 85
90 95 Met Asp Arg Met Asn Lys Ser Ala Asp Arg Ile
Ala Leu Pro Thr Phe 100 105
110 Asp Gly Glu Glu Leu Met Lys Leu Ile Asp Gln Phe Leu Leu Val
Asp 115 120 125 Glu
Ser Phe Val Pro Gln Gly Ala Gly Tyr Ser Leu Tyr Leu Arg Pro 130
135 140 Thr Met Ile Gly Thr Thr
Glu Ser Leu Gly Val Gly Thr Pro Asp Lys 145 150
155 160 Ala Leu Leu Tyr Val Ile Ala Ser Pro Val Gly
Pro Tyr Tyr Gly Thr 165 170
175 Gly Phe Lys Pro Val Ser Leu Glu Ala Thr Asp Tyr Ala Val Arg Ala
180 185 190 Trp Pro
Gly Gly Val Gly Asn Arg Lys Leu Gly Ala Asn Tyr Ala Pro 195
200 205 Cys Val Arg Pro Gln Leu Glu
Ala Ala Lys Arg Gly Tyr Gln Gln Asn 210 215
220 Leu Trp Leu Phe Gly Glu Glu Gly Tyr Ile Thr Glu
Val Gly Thr Met 225 230 235
240 Asn Ala Phe Phe Val Phe Lys Asn Ala Asp Gly Thr Lys Glu Leu Ala
245 250 255 Thr Ala Pro
Leu Asp Gly Thr Ile Leu Glu Gly Val Thr Arg Asp Ser 260
265 270 Ile Leu Glu Leu Thr Arg Glu Arg
Leu Pro Lys Asn Glu Trp Val Val 275 280
285 Ser Glu Arg Lys Phe Thr Ile Gly Glu Val Glu Glu Arg
Ala Ala Lys 290 295 300
Gly Glu Leu Ile Glu Ala Phe Gly Ala Gly Thr Ala Ala Val Val Ser 305
310 315 320 Pro Ile Lys Ser
Ile Gly Trp Lys Gly Lys Glu Ile Glu Val Pro Leu 325
330 335 Ala Ala Gly Asp Ser Gly Glu Leu Thr
Ala Gln Val Ala Glu Trp Ile 340 345
350 Arg Lys Ile Gln Tyr Gly Glu Glu Gln Tyr Lys Asn Trp Ser
Arg Val 355 360 365
Ala Gln 370 271860DNASaccharomyces cerevisiae 27atggttaaag agagtattat
tgctcttgct gagcatgcgg cctccagagc ctcaagagtt 60attcctccag tgaagctagc
ctataaaaat atgcttaagg acccttcctc caagtacaag 120ccatttaacg ctccaaagct
atctaataga aagtggccgg ataaccggat cacgagggct 180cctcgttggt tatcaacaga
tttgagagat ggtaaccaat ctctgccgga tcccatgtca 240gtggaacaaa agaaagaata
ctttcacaag ctggtcaata ttgggttcaa agaaatcgag 300gtttccttcc cctctgcatc
tcaaacagat ttcgacttca ctagatatgc tgtagaaaac 360gccccagacg atgttagtat
tcaatgtctt gtccaatcta gagaacactt gattaagaga 420acggtggaag cattaacagg
tgctaaaaag gctactatac atacttactt ggcaacaagt 480gatatgttcc gtgaaattgt
ttttaatatg tctagagagg aagctatttc caaggcagta 540gaggccacca aactagttag
gaaactaact aaggatgacc cttcccaaca agccactcgt 600tggtcctatg agttttcccc
cgaatgtttc agtgatactc caggtgaatt tgctgtagaa 660atttgcgaag ctgttaagaa
ggcttgggaa cctaccgagg aaaatccaat cattttcaac 720ttacctgcta ccgtagaagt
tgcctctcca aatgtttatg ctgatcagat tgaatacttc 780gctacccata ttactgagcg
tgagaaggtt tgcatctcta cacattgtca caatgaccgt 840ggttgcggtg tcgccgccac
agagttaggt atgcttgcag gtgccgaccg tgtagaagga 900tgtctctttg gtaatggtga
acgtacaggt aatgtggact tggttactgt tgctatgaat 960atgtataccc aaggtgtttc
tcctaatttg gatttctctg acttgacctc tgtcctagat 1020gtggttgagc gttgtaataa
gatcccagta tcgcaaagag caccatacgg cggtgacttg 1080gtcgtttgtg ccttttccgg
ttctcaccaa gacgccatta agaagggttt caacttacaa 1140aacaagaagc gtgctcaagg
tgaaactcaa tggagaatcc catacttgcc attggatcca 1200aaggacattg gccgtgatta
cgaagctgtc atcagagtca actctcagtc tggtaaaggt 1260ggtgccgctt gggttatttt
gagatctttg ggtttggatc taccaagaaa catgcaaatc 1320gaattttcta gcgccgttca
agaccatgct gactccttgg gtagagaact aaaatcagat 1380gagatttcca agttattcaa
agaggcttac aactacaatg acgaacagta ccaagctatt 1440agtttagtca attataatgt
tgaaaaattc ggcactgaac gtagagtgtt cactggtcaa 1500gtcaaagtag gcgaccagat
cgtcgatatt gaaggtacag gtaatggtcc aatctcttct 1560ttggtcgacg ccctatcaaa
cttgttgaac gtgagatttg ccgtagcaaa ctacacagag 1620cattctctag gttctggttc
ttctacgcaa gctgcttctt acatccatct atcgtatagg 1680cgtaatgccg acaacgaaaa
ggcctacaaa tggggtgtag gtgtctccga agatgtcggt 1740gattcttcag tgagagccat
ctttgccacc attaacaata ttatccattc tggtgatgtg 1800tccattccat ctttggccga
ggtcgaaggt aagaatgctg cggcatctgg ctctgcataa 186028619PRTSaccharomyces
cerevisiae 28Met Val Lys Glu Ser Ile Ile Ala Leu Ala Glu His Ala Ala Ser
Arg 1 5 10 15 Ala
Ser Arg Val Ile Pro Pro Val Lys Leu Ala Tyr Lys Asn Met Leu
20 25 30 Lys Asp Pro Ser Ser
Lys Tyr Lys Pro Phe Asn Ala Pro Lys Leu Ser 35
40 45 Asn Arg Lys Trp Pro Asp Asn Arg Ile
Thr Arg Ala Pro Arg Trp Leu 50 55
60 Ser Thr Asp Leu Arg Asp Gly Asn Gln Ser Leu Pro Asp
Pro Met Ser 65 70 75
80 Val Glu Gln Lys Lys Glu Tyr Phe His Lys Leu Val Asn Ile Gly Phe
85 90 95 Lys Glu Ile Glu
Val Ser Phe Pro Ser Ala Ser Gln Thr Asp Phe Asp 100
105 110 Phe Thr Arg Tyr Ala Val Glu Asn Ala
Pro Asp Asp Val Ser Ile Gln 115 120
125 Cys Leu Val Gln Ser Arg Glu His Leu Ile Lys Arg Thr Val
Glu Ala 130 135 140
Leu Thr Gly Ala Lys Lys Ala Thr Ile His Thr Tyr Leu Ala Thr Ser 145
150 155 160 Asp Met Phe Arg Glu
Ile Val Phe Asn Met Ser Arg Glu Glu Ala Ile 165
170 175 Ser Lys Ala Val Glu Ala Thr Lys Leu Val
Arg Lys Leu Thr Lys Asp 180 185
190 Asp Pro Ser Gln Gln Ala Thr Arg Trp Ser Tyr Glu Phe Ser Pro
Glu 195 200 205 Cys
Phe Ser Asp Thr Pro Gly Glu Phe Ala Val Glu Ile Cys Glu Ala 210
215 220 Val Lys Lys Ala Trp Glu
Pro Thr Glu Glu Asn Pro Ile Ile Phe Asn 225 230
235 240 Leu Pro Ala Thr Val Glu Val Ala Ser Pro Asn
Val Tyr Ala Asp Gln 245 250
255 Ile Glu Tyr Phe Ala Thr His Ile Thr Glu Arg Glu Lys Val Cys Ile
260 265 270 Ser Thr
His Cys His Asn Asp Arg Gly Cys Gly Val Ala Ala Thr Glu 275
280 285 Leu Gly Met Leu Ala Gly Ala
Asp Arg Val Glu Gly Cys Leu Phe Gly 290 295
300 Asn Gly Glu Arg Thr Gly Asn Val Asp Leu Val Thr
Val Ala Met Asn 305 310 315
320 Met Tyr Thr Gln Gly Val Ser Pro Asn Leu Asp Phe Ser Asp Leu Thr
325 330 335 Ser Val Leu
Asp Val Val Glu Arg Cys Asn Lys Ile Pro Val Ser Gln 340
345 350 Arg Ala Pro Tyr Gly Gly Asp Leu
Val Val Cys Ala Phe Ser Gly Ser 355 360
365 His Gln Asp Ala Ile Lys Lys Gly Phe Asn Leu Gln Asn
Lys Lys Arg 370 375 380
Ala Gln Gly Glu Thr Gln Trp Arg Ile Pro Tyr Leu Pro Leu Asp Pro 385
390 395 400 Lys Asp Ile Gly
Arg Asp Tyr Glu Ala Val Ile Arg Val Asn Ser Gln 405
410 415 Ser Gly Lys Gly Gly Ala Ala Trp Val
Ile Leu Arg Ser Leu Gly Leu 420 425
430 Asp Leu Pro Arg Asn Met Gln Ile Glu Phe Ser Ser Ala Val
Gln Asp 435 440 445
His Ala Asp Ser Leu Gly Arg Glu Leu Lys Ser Asp Glu Ile Ser Lys 450
455 460 Leu Phe Lys Glu Ala
Tyr Asn Tyr Asn Asp Glu Gln Tyr Gln Ala Ile 465 470
475 480 Ser Leu Val Asn Tyr Asn Val Glu Lys Phe
Gly Thr Glu Arg Arg Val 485 490
495 Phe Thr Gly Gln Val Lys Val Gly Asp Gln Ile Val Asp Ile Glu
Gly 500 505 510 Thr
Gly Asn Gly Pro Ile Ser Ser Leu Val Asp Ala Leu Ser Asn Leu 515
520 525 Leu Asn Val Arg Phe Ala
Val Ala Asn Tyr Thr Glu His Ser Leu Gly 530 535
540 Ser Gly Ser Ser Thr Gln Ala Ala Ser Tyr Ile
His Leu Ser Tyr Arg 545 550 555
560 Arg Asn Ala Asp Asn Glu Lys Ala Tyr Lys Trp Gly Val Gly Val Ser
565 570 575 Glu Asp
Val Gly Asp Ser Ser Val Arg Ala Ile Phe Ala Thr Ile Asn 580
585 590 Asn Ile Ile His Ser Gly Asp
Val Ser Ile Pro Ser Leu Ala Glu Val 595 600
605 Glu Gly Lys Asn Ala Ala Ala Ser Gly Ser Ala
610 615 291755DNASchizosaccharomyces
pombe 29atgaaatcta cttttgaggc tgctggccgc gttgccaaag ggatgctcaa ggatccctcc
60aaaaagtata agccatttaa aggaattcaa ctacccaacc gtcaatggcc aaacaaggtt
120ttgacgaaag ctccacgctg gctttctacg gacttgcgtg atggtaatca ggctttaccc
180gatcctatga atgggcagga gaaattgaga tattttaaat tgctttgcag tattggcttc
240aaagaaattg aggttggttt cccaagtgct tctcaaactg attttgcatt tgttcgtcat
300ctgattgaaa cgccaggttt gattcctgac gatgttacta tttctgccct tactccttct
360cgtgagcctt tgatcctacg tacgattgag gctcttcgag gcgctaagaa tgccactgtt
420cacttgtata atgcctgttc tcctcttttc cgtgaagttg tcttccgcaa cagtaagcaa
480gaaacattgg atttagccat caaaggctca aaaatcgtaa cagctgctac gaaaaatgct
540cttgaatcga aggaaaccaa ctggggattt gaatattctc ctgaaacttt ttcagacacc
600gaaccagact ttgctttgga agtttgtgaa gctgtcaagg gtatgtggaa accttctgct
660gcccaaccta ttatcttcaa tcttcctgcc actgtcgaaa tgtctacgcc caacacatat
720gctgacttaa ttgagtactt ttccactaac attagtgaac gtgaaaaagt ctgtgtttct
780ctccatcccc ataacgaccg tggtactgct gtcgcagcag ctgaacttgg tcaacttgcc
840ggaggtgacc gtattgaggg ctgtttgttt ggcaatggtg aacgtactgg taatgtagac
900ttggttactt tggctttcaa cttgtatacc caaggtgttt ctcctaacct cgatttctcc
960aagttggatg aaatcattcg tattactgaa gactgtaaca agataaacgt tcatccccgt
1020catccttatg ctggcaatct tgtctttacc gccttttctg gttctcatca agatgccatt
1080tctaagggtt tgaaggctta cgatgagcgt aaagctgtcg atcctgtttg gaaagtccct
1140tacttgcctt tggatcccca tgatgtcaat tccgagtatg ctgctattat ccgcgttaac
1200tctcaatctg gcaagggtgg tgtcgcatat ctgttgaaga ccaactgtgg tctcgattta
1260cctcgtgctt tgcaagttga atttggtagt attgttaagg attatagcga cacaaaagga
1320aaggagctta gcattggtga gatcagcgac ctgttttata ccacatatta cctcgaattt
1380cccggccgtt tctctgtaaa cgactacact ctttctagca acggacctca aagcaaatgt
1440attaaatgcg ttgttgacat caagggtgaa aagaaagata ctccttcgcg ggttgtgatc
1500gagggtgttg gaaatggacc tttgtcggca ttggttgatg ctttacgccg tcagttcaat
1560atttcatttg acattggtca atactctgaa catgctattg gttctggtaa cggcgtcaaa
1620gctgcttctt atgttgagat cattttcaat aacacttctt tctggggtgt tggtattgat
1680gctgacgtta cctctgccgg attaaaggct gtcatgtcag gcgttagtcg tgcctcccgc
1740gcatttgcta agtaa
175530584PRTSchizosaccharomyces pombe 30Met Lys Ser Thr Phe Glu Ala Ala
Gly Arg Val Ala Lys Gly Met Leu 1 5 10
15 Lys Asp Pro Ser Lys Lys Tyr Lys Pro Phe Lys Gly Ile
Gln Leu Pro 20 25 30
Asn Arg Gln Trp Pro Asn Lys Val Leu Thr Lys Ala Pro Arg Trp Leu
35 40 45 Ser Thr Asp Leu
Arg Asp Gly Asn Gln Ala Leu Pro Asp Pro Met Asn 50
55 60 Gly Gln Glu Lys Leu Arg Tyr Phe
Lys Leu Leu Cys Ser Ile Gly Phe 65 70
75 80 Lys Glu Ile Glu Val Gly Phe Pro Ser Ala Ser Gln
Thr Asp Phe Ala 85 90
95 Phe Val Arg His Leu Ile Glu Thr Pro Gly Leu Ile Pro Asp Asp Val
100 105 110 Thr Ile Ser
Ala Leu Thr Pro Ser Arg Glu Pro Leu Ile Leu Arg Thr 115
120 125 Ile Glu Ala Leu Arg Gly Ala Lys
Asn Ala Thr Val His Leu Tyr Asn 130 135
140 Ala Cys Ser Pro Leu Phe Arg Glu Val Val Phe Arg Asn
Ser Lys Gln 145 150 155
160 Glu Thr Leu Asp Leu Ala Ile Lys Gly Ser Lys Ile Val Thr Ala Ala
165 170 175 Thr Lys Asn Ala
Leu Glu Ser Lys Glu Thr Asn Trp Gly Phe Glu Tyr 180
185 190 Ser Pro Glu Thr Phe Ser Asp Thr Glu
Pro Asp Phe Ala Leu Glu Val 195 200
205 Cys Glu Ala Val Lys Gly Met Trp Lys Pro Ser Ala Ala Gln
Pro Ile 210 215 220
Ile Phe Asn Leu Pro Ala Thr Val Glu Met Ser Thr Pro Asn Thr Tyr 225
230 235 240 Ala Asp Leu Ile Glu
Tyr Phe Ser Thr Asn Ile Ser Glu Arg Glu Lys 245
250 255 Val Cys Val Ser Leu His Pro His Asn Asp
Arg Gly Thr Ala Val Ala 260 265
270 Ala Ala Glu Leu Gly Gln Leu Ala Gly Gly Asp Arg Ile Glu Gly
Cys 275 280 285 Leu
Phe Gly Asn Gly Glu Arg Thr Gly Asn Val Asp Leu Val Thr Leu 290
295 300 Ala Phe Asn Leu Tyr Thr
Gln Gly Val Ser Pro Asn Leu Asp Phe Ser 305 310
315 320 Lys Leu Asp Glu Ile Ile Arg Ile Thr Glu Asp
Cys Asn Lys Ile Asn 325 330
335 Val His Pro Arg His Pro Tyr Ala Gly Asn Leu Val Phe Thr Ala Phe
340 345 350 Ser Gly
Ser His Gln Asp Ala Ile Ser Lys Gly Leu Lys Ala Tyr Asp 355
360 365 Glu Arg Lys Ala Val Asp Pro
Val Trp Lys Val Pro Tyr Leu Pro Leu 370 375
380 Asp Pro His Asp Val Asn Ser Glu Tyr Ala Ala Ile
Ile Arg Val Asn 385 390 395
400 Ser Gln Ser Gly Lys Gly Gly Val Ala Tyr Leu Leu Lys Thr Asn Cys
405 410 415 Gly Leu Asp
Leu Pro Arg Ala Leu Gln Val Glu Phe Gly Ser Ile Val 420
425 430 Lys Asp Tyr Ser Asp Thr Lys Gly
Lys Glu Leu Ser Ile Gly Glu Ile 435 440
445 Ser Asp Leu Phe Tyr Thr Thr Tyr Tyr Leu Glu Phe Pro
Gly Arg Phe 450 455 460
Ser Val Asn Asp Tyr Thr Leu Ser Ser Asn Gly Pro Gln Ser Lys Cys 465
470 475 480 Ile Lys Cys Val
Val Asp Ile Lys Gly Glu Lys Lys Asp Thr Pro Ser 485
490 495 Arg Val Val Ile Glu Gly Val Gly Asn
Gly Pro Leu Ser Ala Leu Val 500 505
510 Asp Ala Leu Arg Arg Gln Phe Asn Ile Ser Phe Asp Ile Gly
Gln Tyr 515 520 525
Ser Glu His Ala Ile Gly Ser Gly Asn Gly Val Lys Ala Ala Ser Tyr 530
535 540 Val Glu Ile Ile Phe
Asn Asn Thr Ser Phe Trp Gly Val Gly Ile Asp 545 550
555 560 Ala Asp Val Thr Ser Ala Gly Leu Lys Ala
Val Met Ser Gly Val Ser 565 570
575 Arg Ala Ser Arg Ala Phe Ala Lys 580
31 1257DNASchizosaccharomyces pombe 31atgtctgtgt ccgaagctaa
tggtactgag accatcaagc ctcctatgaa tggaaaccct 60tatggtccca acccatctga
ttttctttca cgtgtcaata acttttccat tattgagtct 120actcttcgtg aaggtgagca
attcgcaaac gcttttttcg acaccgagaa gaaaattcaa 180attgctaagg cattggacaa
ctttggtgtc gattacattg aattgacttc tcccgtggct 240tctgagcagt cccgccaaga
ttgcgaagct atttgcaaat tgggcttaaa gtgtaaaatt 300ttaactcata ttcgctgtca
tatggatgac gctcgtgtcg ctgttgagac tggagttgat 360ggtgttgatg ttgttatcgg
aacttctcaa tatcttcgca aatattccca tggaaaggac 420atgacttaca ttattgacag
cgctaccgaa gttatcaact ttgtcaagag caagggtatt 480gaagtccgct tttcatctga
ggattctttc cgttctgatc ttgtcgatct cctttctctc 540tacaaggctg tagacaagat
tggcgtcaac cgtgttggta ttgctgacac cgttggttgc 600gctactcctc gccaagtcta
cgatcttatt cgtaccttac gtggtgttgt ctcttgtgat 660attgaatgtc attttcacaa
tgacactggt atggctattg ctaatgccta ttgcgcattg 720gaagctggtg ctacccatat
cgatacttcc attcttggta ttggtgagcg taatggtatt 780actcctcttg gtgccttgtt
ggctcgtatg tatgtcaccg atagggaata cattacccac 840aaatacaagc ttaaccagtt
acgtgagctt gaaaaccttg tcgctgatgc cgttgaagtt 900caaattcctt tcaacaatta
cattaccgga atgtgtgctt ttacccataa ggctggtatc 960catgctaaag ctattctcgc
taacccttct acatatgaaa ttcttaagcc cgaggacttt 1020ggcatgagtc gttatgttca
tgttggctct cgtttgactg gttggaatgc catcaaatct 1080cgtgctgagc agcttaacct
tcatcttact gatgcccaag ccaaggaact taccgttcgc 1140atcaagaaat tggctgatgt
ccgtacttta gccatggatg atgttgatcg tgttctacgt 1200gaataccatg ctgacttgag
tgatgctgat agaatcacca aagaagcgtc tgcttaa
125732418PRTSchizosaccharomyces pombe 32Met Ser Val Ser Glu Ala Asn Gly
Thr Glu Thr Ile Lys Pro Pro Met 1 5 10
15 Asn Gly Asn Pro Tyr Gly Pro Asn Pro Ser Asp Phe Leu
Ser Arg Val 20 25 30
Asn Asn Phe Ser Ile Ile Glu Ser Thr Leu Arg Glu Gly Glu Gln Phe
35 40 45 Ala Asn Ala Phe
Phe Asp Thr Glu Lys Lys Ile Gln Ile Ala Lys Ala 50
55 60 Leu Asp Asn Phe Gly Val Asp Tyr
Ile Glu Leu Thr Ser Pro Val Ala 65 70
75 80 Ser Glu Gln Ser Arg Gln Asp Cys Glu Ala Ile Cys
Lys Leu Gly Leu 85 90
95 Lys Cys Lys Ile Leu Thr His Ile Arg Cys His Met Asp Asp Ala Arg
100 105 110 Val Ala Val
Glu Thr Gly Val Asp Gly Val Asp Val Val Ile Gly Thr 115
120 125 Ser Gln Tyr Leu Arg Lys Tyr Ser
His Gly Lys Asp Met Thr Tyr Ile 130 135
140 Ile Asp Ser Ala Thr Glu Val Ile Asn Phe Val Lys Ser
Lys Gly Ile 145 150 155
160 Glu Val Arg Phe Ser Ser Glu Asp Ser Phe Arg Ser Asp Leu Val Asp
165 170 175 Leu Leu Ser Leu
Tyr Lys Ala Val Asp Lys Ile Gly Val Asn Arg Val 180
185 190 Gly Ile Ala Asp Thr Val Gly Cys Ala
Thr Pro Arg Gln Val Tyr Asp 195 200
205 Leu Ile Arg Thr Leu Arg Gly Val Val Ser Cys Asp Ile Glu
Cys His 210 215 220
Phe His Asn Asp Thr Gly Met Ala Ile Ala Asn Ala Tyr Cys Ala Leu 225
230 235 240 Glu Ala Gly Ala Thr
His Ile Asp Thr Ser Ile Leu Gly Ile Gly Glu 245
250 255 Arg Asn Gly Ile Thr Pro Leu Gly Ala Leu
Leu Ala Arg Met Tyr Val 260 265
270 Thr Asp Arg Glu Tyr Ile Thr His Lys Tyr Lys Leu Asn Gln Leu
Arg 275 280 285 Glu
Leu Glu Asn Leu Val Ala Asp Ala Val Glu Val Gln Ile Pro Phe 290
295 300 Asn Asn Tyr Ile Thr Gly
Met Cys Ala Phe Thr His Lys Ala Gly Ile 305 310
315 320 His Ala Lys Ala Ile Leu Ala Asn Pro Ser Thr
Tyr Glu Ile Leu Lys 325 330
335 Pro Glu Asp Phe Gly Met Ser Arg Tyr Val His Val Gly Ser Arg Leu
340 345 350 Thr Gly
Trp Asn Ala Ile Lys Ser Arg Ala Glu Gln Leu Asn Leu His 355
360 365 Leu Thr Asp Ala Gln Ala Lys
Glu Leu Thr Val Arg Ile Lys Lys Leu 370 375
380 Ala Asp Val Arg Thr Leu Ala Met Asp Asp Val Asp
Arg Val Leu Arg 385 390 395
400 Glu Tyr His Ala Asp Leu Ser Asp Ala Asp Arg Ile Thr Lys Glu Ala
405 410 415 Ser Ala
331740DNACandida albicans 33atgcaaaagg ttttccaaag atgggtatct agaatacccc
cagttaagct ccaatataag 60aatatgctta gagacccttc caaaaaatac tctccaccaa
aacagatcaa cttgcccaat 120agaacttggc ccaccaaagt aatcactaaa gctccccgct
ggctttccac tgatttaaga 180gacggtaacc agtccttgcc agatccaatg tcggttccag
aaaaaaaaga atacttccat 240aaattaattg atattgggtt taaagaaatc gaagtttcgt
tcccctctgc ttcgcaaact 300gattttgatt tcacccgata cgccgttgaa aatgcgccag
atgatgtaac tattcaagtc 360ttgacccaat ctcgtgaacc attgatcaga agaacagtgg
aatcggtaaa aggggccaag 420cgtgctacca ttcatacata tttggcaacc tctgatgtat
tccgtgaagt tgttttcggt 480atgagcaaac aagacgctat agacaaggcc attgaaacta
caaaattagt gagatcatta 540actaaagatg accctaacat gcaagacact gaatggaatt
tggagttttc tccagagtgt 600ttctcagata cgccagttga atttgccgtt gagatttgtg
aagccgttaa aaaagcttgg 660gaaccaacag tggaaaaccc aatgatcttc aatttgcctg
ccacagttga agttgctggt 720cctaatgttt atgctgatca gattgaatac ttttgtcaaa
acataactga acgtgaaaag 780attattgtct ccacccatac tcataatgac cgtggctgtg
gtgtcgctgc taccgaattg 840ggtatgttgg ctggtgccga tagagtggaa ggttgtgtgt
ttggaaacgg tgaaagaacc 900ggtaatgttg acttggtcac ggtggcattg aacttgtaca
cccaaggtat tgcgccaaat 960ttggactttt ccgatatcga gagcattatt gaggttagtg
aacgttgtaa taaaatcccg 1020gtgcccgcaa gatcacctta cggtggctca cttgtggtgt
gtgccttcag tggatctcat 1080caagacgcca ttaaaaaggg ttttgctaaa caaaagggag
acaaatgggc tatcccatac 1140ttgccattag atccaaaaga tattggcaga acttacgaag
ccgtgattag agtcaactcc 1200caatcaggta aaggtggtgc tgcctgggtc atccttagat
ctttgggatt ggacttgcca 1260agacacttac aagttgcctt ttcaggattg gtgcaaaaca
ctgctgacct gttgggtaga 1320gaattgaagg ttgatgaaat tgtcaacttg ttcaacgaac
aatacttggt gagtgcccct 1380ttaagcattc aggattttga aatcaccaag aataaaaacg
atgaaagaga aattgttgct 1440caattaaatg atggcatcac cattaaaggt caaggtaatg
gtcctatctc tgcttttatt 1500gatgcaattt ctaacaagtt cggtgttttg tttgaagttg
taaactatca agaacattct 1560ttgggaggtg gttctagtag taaggcagca acttatatcg
aattatcata tgttaatgcc 1620aatggtgaaa aagttactag atggggttgt ggtatcaatc
acgatgtgtc acaagcctca 1680atcgaagcca ttcttagtgt tgtaaactct ttgattaaaa
agaatgaatt aactgtatag 174034579PRTCandida albicans 34Met Gln Lys Val
Phe Gln Arg Trp Val Ser Arg Ile Pro Pro Val Lys 1 5
10 15 Leu Gln Tyr Lys Asn Met Leu Arg Asp
Pro Ser Lys Lys Tyr Ser Pro 20 25
30 Pro Lys Gln Ile Asn Leu Pro Asn Arg Thr Trp Pro Thr Lys
Val Ile 35 40 45
Thr Lys Ala Pro Arg Trp Leu Ser Thr Asp Leu Arg Asp Gly Asn Gln 50
55 60 Ser Leu Pro Asp Pro
Met Ser Val Pro Glu Lys Lys Glu Tyr Phe His 65 70
75 80 Lys Leu Ile Asp Ile Gly Phe Lys Glu Ile
Glu Val Ser Phe Pro Ser 85 90
95 Ala Ser Gln Thr Asp Phe Asp Phe Thr Arg Tyr Ala Val Glu Asn
Ala 100 105 110 Pro
Asp Asp Val Thr Ile Gln Val Leu Thr Gln Ser Arg Glu Pro Leu 115
120 125 Ile Arg Arg Thr Val Glu
Ser Val Lys Gly Ala Lys Arg Ala Thr Ile 130 135
140 His Thr Tyr Leu Ala Thr Ser Asp Val Phe Arg
Glu Val Val Phe Gly 145 150 155
160 Met Ser Lys Gln Asp Ala Ile Asp Lys Ala Ile Glu Thr Thr Lys Leu
165 170 175 Val Arg
Ser Leu Thr Lys Asp Asp Pro Asn Met Gln Asp Thr Glu Trp 180
185 190 Asn Leu Glu Phe Ser Pro Glu
Cys Phe Ser Asp Thr Pro Val Glu Phe 195 200
205 Ala Val Glu Ile Cys Glu Ala Val Lys Lys Ala Trp
Glu Pro Thr Val 210 215 220
Glu Asn Pro Met Ile Phe Asn Leu Pro Ala Thr Val Glu Val Ala Gly 225
230 235 240 Pro Asn Val
Tyr Ala Asp Gln Ile Glu Tyr Phe Cys Gln Asn Ile Thr 245
250 255 Glu Arg Glu Lys Ile Ile Val Ser
Thr His Thr His Asn Asp Arg Gly 260 265
270 Cys Gly Val Ala Ala Thr Glu Leu Gly Met Leu Ala Gly
Ala Asp Arg 275 280 285
Val Glu Gly Cys Val Phe Gly Asn Gly Glu Arg Thr Gly Asn Val Asp 290
295 300 Leu Val Thr Val
Ala Leu Asn Leu Tyr Thr Gln Gly Ile Ala Pro Asn 305 310
315 320 Leu Asp Phe Ser Asp Ile Glu Ser Ile
Ile Glu Val Ser Glu Arg Cys 325 330
335 Asn Lys Ile Pro Val Pro Ala Arg Ser Pro Tyr Gly Gly Ser
Leu Val 340 345 350
Val Cys Ala Phe Ser Gly Ser His Gln Asp Ala Ile Lys Lys Gly Phe
355 360 365 Ala Lys Gln Lys
Gly Asp Lys Trp Ala Ile Pro Tyr Leu Pro Leu Asp 370
375 380 Pro Lys Asp Ile Gly Arg Thr Tyr
Glu Ala Val Ile Arg Val Asn Ser 385 390
395 400 Gln Ser Gly Lys Gly Gly Ala Ala Trp Val Ile Leu
Arg Ser Leu Gly 405 410
415 Leu Asp Leu Pro Arg His Leu Gln Val Ala Phe Ser Gly Leu Val Gln
420 425 430 Asn Thr Ala
Asp Ser Leu Gly Arg Glu Leu Lys Val Asp Glu Ile Val 435
440 445 Asn Leu Phe Asn Glu Gln Tyr Leu
Val Ser Ala Pro Leu Ser Ile Gln 450 455
460 Asp Phe Glu Ile Thr Lys Asn Lys Asn Asp Glu Arg Glu
Ile Val Ala 465 470 475
480 Gln Leu Asn Asp Gly Ile Thr Ile Lys Gly Gln Gly Asn Gly Pro Ile
485 490 495 Ser Ala Phe Ile
Asp Ala Ile Ser Asn Lys Phe Gly Val Leu Phe Glu 500
505 510 Val Val Asn Tyr Gln Glu His Ser Leu
Gly Gly Gly Ser Ser Ser Lys 515 520
525 Ala Ala Thr Tyr Ile Glu Leu Ser Tyr Val Asn Ala Asn Gly
Glu Lys 530 535 540
Val Thr Arg Trp Gly Cys Gly Ile Asn His Asp Val Ser Gln Ala Ser 545
550 555 560 Ile Glu Ala Ile Leu
Ser Val Val Asn Ser Leu Ile Lys Lys Asn Glu 565
570 575 Leu Thr Val 351722DNACandida albicans
35atgcctatgt taaaagatcc ctcagtgaaa tataagaagt ttccaaatgt caatttgcca
60aaccgtcaat ggccatcaag aagcttggat aaaccaccaa gatggttatc tactgatttg
120agagatggta accaatcatt acctgatcca atgtcgatct ctgaaaagaa agaatatttc
180aagaaattgg ttgatatagg attcaaagaa atcgaagttg ccttcccctc agcctctcaa
240attgatttcg atttcactag atttgccgtt gaaactgccc ctgaagatgt ttcgattcaa
300gttctttctc catgtcgtcc cgaattgatt ggtagaactg ttgaatcttt gaaaggtgct
360aaaagagcaa ctgtccacat atatcttgcc acttctgatt gttttagaaa tgttgtgttt
420ggactttcca aagaagaatc aaaggcctta gctgtgaaat gtaccaaatt ggtgagacaa
480ttaactaaag atgatccttc aactgccggt acagattggg attttgaatt ttctccagaa
540actttttctg acacagattt ggattatgct gttgaagtat gtgaagcagt caaagaagcc
600tgggggccaa cagaagataa accaattata tttaatttgc cagcaactgt tgaaatggcc
660actcctaaca tatatgctga tcaaattgaa tattttgcca ctcatattac tgaccgtgaa
720acagtttgta tttcattgca tcctcacaat gatagagggt gtagtgttgc tgctgccgaa
780ttaggtcaat tagctggtgc tgacagagtt gaaggttgtc ttttcggtaa tggtgaaaga
840accggtaatg ttgatttagt cactttagca ttgaacttgt atacccaggg ggtatcacca
900aaattggact tttctgattt gaattcggtc attgatatag ttgaaaaatg caacaaaatt
960cctgttcatg ctagagctcc atacggaggg tctcttgttg tttgtgcctt tagtggatct
1020catcaagatg ccatcaaaaa ggggttcctg gctcacgaaa agaaaaaaga aaaagcggga
1080ggcaaagaag ttcattggca attaccttat ttaccattgg atccagaaga tattggaaga
1140acatacgagg ctattattag agtgaattct caatctggta aaggtggttc tgcttgggtg
1200atcttgagaa atttggaatt agatttacct cgtggtttac aaattgcctt ctctaaagtg
1260gttcaagcac gtgctgaagt taaaggtcaa gaattaacta acgaagaatt atgtgagtta
1320ttcaagcaag aatatttcat tgattatgat gatgaagccc cagaacaata ctttaaatta
1380gtagattact cgatatcgac accaagcaaa ggaatcaagg aaatccaagc tgatattgaa
1440gtcgatggta aagtcatttc tatcaaaggt gaaggtaatg gtcaattatc tgcctttaat
1500aatgccattg ctaaatattt gaatattgat attgacgtga aacattatca cgaacattcc
1560cttggtgaag attcaaaagc ccgtgccgcc acttatattg aagtcttggt cgataaaaaa
1620gttgcaagat ggggtgtggg tattcatact gatgtttctc aagcttcatt cttatctttg
1680atatctattt tgaatggttt gcataaaaat aaaaacattt aa
172236573PRTCandida albicans 36Met Pro Met Leu Lys Asp Pro Ser Val Lys
Tyr Lys Lys Phe Pro Asn 1 5 10
15 Val Asn Leu Pro Asn Arg Gln Trp Pro Ser Arg Ser Leu Asp Lys
Pro 20 25 30 Pro
Arg Trp Leu Ser Thr Asp Leu Arg Asp Gly Asn Gln Ser Leu Pro 35
40 45 Asp Pro Met Ser Ile Ser
Glu Lys Lys Glu Tyr Phe Lys Lys Leu Val 50 55
60 Asp Ile Gly Phe Lys Glu Ile Glu Val Ala Phe
Pro Ser Ala Ser Gln 65 70 75
80 Ile Asp Phe Asp Phe Thr Arg Phe Ala Val Glu Thr Ala Pro Glu Asp
85 90 95 Val Ser
Ile Gln Val Leu Ser Pro Cys Arg Pro Glu Leu Ile Gly Arg 100
105 110 Thr Val Glu Ser Leu Lys Gly
Ala Lys Arg Ala Thr Val His Ile Tyr 115 120
125 Leu Ala Thr Ser Asp Cys Phe Arg Asn Val Val Phe
Gly Leu Ser Lys 130 135 140
Glu Glu Ser Lys Ala Leu Ala Val Lys Cys Thr Lys Leu Val Arg Gln 145
150 155 160 Leu Thr Lys
Asp Asp Pro Ser Thr Ala Gly Thr Asp Trp Asp Phe Glu 165
170 175 Phe Ser Pro Glu Thr Phe Ser Asp
Thr Asp Leu Asp Tyr Ala Val Glu 180 185
190 Val Cys Glu Ala Val Lys Glu Ala Trp Gly Pro Thr Glu
Asp Lys Pro 195 200 205
Ile Ile Phe Asn Leu Pro Ala Thr Val Glu Met Ala Thr Pro Asn Ile 210
215 220 Tyr Ala Asp Gln
Ile Glu Tyr Phe Ala Thr His Ile Thr Asp Arg Glu 225 230
235 240 Thr Val Cys Ile Ser Leu His Pro His
Asn Asp Arg Gly Cys Ser Val 245 250
255 Ala Ala Ala Glu Leu Gly Gln Leu Ala Gly Ala Asp Arg Val
Glu Gly 260 265 270
Cys Leu Phe Gly Asn Gly Glu Arg Thr Gly Asn Val Asp Leu Val Thr
275 280 285 Leu Ala Leu Asn
Leu Tyr Thr Gln Gly Val Ser Pro Lys Leu Asp Phe 290
295 300 Ser Asp Leu Asn Ser Val Ile Asp
Ile Val Glu Lys Cys Asn Lys Ile 305 310
315 320 Pro Val His Ala Arg Ala Pro Tyr Gly Gly Ser Leu
Val Val Cys Ala 325 330
335 Phe Ser Gly Ser His Gln Asp Ala Ile Lys Lys Gly Phe Ser Ala His
340 345 350 Glu Lys Lys
Lys Glu Lys Ala Gly Gly Lys Glu Val His Trp Gln Leu 355
360 365 Pro Tyr Leu Pro Leu Asp Pro Glu
Asp Ile Gly Arg Thr Tyr Glu Ala 370 375
380 Ile Ile Arg Val Asn Ser Gln Ser Gly Lys Gly Gly Ser
Ala Trp Val 385 390 395
400 Ile Leu Arg Asn Leu Glu Leu Asp Leu Pro Arg Gly Leu Gln Ile Ala
405 410 415 Phe Ser Lys Val
Val Gln Ala Arg Ala Glu Val Lys Gly Gln Glu Leu 420
425 430 Thr Asn Glu Glu Leu Cys Glu Leu Phe
Lys Gln Glu Tyr Phe Ile Asp 435 440
445 Tyr Asp Asp Glu Ala Pro Glu Gln Tyr Phe Lys Leu Val Asp
Tyr Ser 450 455 460
Ile Ser Thr Pro Ser Lys Gly Ile Lys Glu Ile Gln Ala Asp Ile Glu 465
470 475 480 Val Asp Gly Lys Val
Ile Ser Ile Lys Gly Glu Gly Asn Gly Gln Leu 485
490 495 Ser Ala Phe Asn Asn Ala Ile Ala Lys Tyr
Leu Asn Ile Asp Ile Asp 500 505
510 Val Lys His Tyr His Glu His Ser Leu Gly Glu Asp Ser Lys Ala
Arg 515 520 525 Ala
Ala Thr Tyr Ile Glu Val Leu Val Asp Lys Lys Val Ala Arg Trp 530
535 540 Gly Val Gly Ile His Thr
Asp Val Ser Gln Ala Ser Phe Leu Ser Leu 545 550
555 560 Ile Ser Ile Leu Asn Gly Leu His Lys Asn Lys
Asn Ile 565 570
371254DNACandida albicans 37atgtctgttg cttctaatcc atatggtcca aatccatctg
atttcttatc taatgtgaat 60aaatttgaag tcattgaatc aactttaaga gaaggtgaac
aatttgccaa tgcctttttc 120accactgaaa aaaaaattga aattgctaaa gctttagatg
attttggggt tgattatatt 180gaattgactt caccagtggc atctgaacaa tcaagaagag
attgtgaagc catttgtaaa 240ttgggtttaa aagccaaaat attgacacat attagatgtc
atatggatga tgcccgtgtt 300gccgttgaaa ctggggttga tggggttgat gtggttattg
gaacttcaca atttttaaga 360caatattctc atggtaaaga tatgaattat attgctcaaa
gtgctattga agtcattgaa 420tttgttaaat ctaaaggtat tgaaattcgt tttagttctg
aagattcttt tagatcagat 480attgttgatt tattaaacat ttatcgtact gttgataaaa
tcggagtgaa tagagttggt 540attgccgata ctgttggttg tgctaaccca agacaagttt
atgaattggt taaaactttg 600aaatcggtgg tttcttgtga tattgaatgt catttccata
acgatactgg ttgtgccatt 660gctaatgctt atactgcctt ggaagccggt gctaaattga
ttgatgtttc tgtgttgggt 720attggtgaaa ggaatggtat tactccattg ggggcattaa
tggcaagaat gattactgct 780gatcgtgatt atgtgttatc taaatataaa ttacacaaat
tgagagattt agaaaatttg 840gttgctgatg ccgtacaaat taatattcca ttcaataatc
caattactgg attctgtgct 900tttactcata aagctggtat tcatgctaaa gccatcttgg
ccaatccatc aacatatgaa 960atcttgaatc caaatgattt cggtttaacc agatatattc
actttgctaa tagattgact 1020ggttggaatg ccattaaatc aagagttgat caattgaatt
tacatttgac tgatgatcaa 1080gttaaagaag ttacaaataa aattaaaaaa ttgggtgatg
ttagacaatt gaacattgat 1140gatgtcgatt caattattaa agatttccat gctgaacaaa
gcactaccaa tactcctctt 1200ttaaaaccag tagaggatga tgaaggtcca gaaattaaaa
aacaaaaagt atag 125438417PRTCandida albicans 38Met Ser Val Ala
Ser Asn Pro Tyr Gly Pro Asn Pro Ser Asp Phe Leu 1 5
10 15 Ser Asn Val Asn Lys Phe Glu Val Ile
Glu Ser Thr Leu Arg Glu Gly 20 25
30 Glu Gln Phe Ala Asn Ala Phe Phe Thr Thr Glu Lys Lys Ile
Glu Ile 35 40 45
Ala Lys Ala Leu Asp Asp Phe Gly Val Asp Tyr Ile Glu Leu Thr Ser 50
55 60 Pro Val Ala Ser Glu
Gln Ser Arg Arg Asp Cys Glu Ala Ile Cys Lys 65 70
75 80 Leu Gly Leu Lys Ala Lys Ile Leu Thr His
Ile Arg Cys His Met Asp 85 90
95 Asp Ala Arg Val Ala Val Glu Thr Gly Val Asp Gly Val Asp Val
Val 100 105 110 Ile
Gly Thr Ser Gln Phe Leu Arg Gln Tyr Ser His Gly Lys Asp Met 115
120 125 Asn Tyr Ile Ala Gln Ser
Ala Ile Glu Val Ile Glu Phe Val Lys Ser 130 135
140 Lys Gly Ile Glu Ile Arg Phe Ser Ser Glu Asp
Ser Phe Arg Ser Asp 145 150 155
160 Ile Val Asp Leu Leu Asn Ile Tyr Arg Thr Val Asp Lys Ile Gly Val
165 170 175 Asn Arg
Val Gly Ile Ala Asp Thr Val Gly Cys Ala Asn Pro Arg Gln 180
185 190 Val Tyr Glu Leu Val Lys Thr
Leu Lys Ser Val Val Ser Cys Asp Ile 195 200
205 Glu Cys His Phe His Asn Asp Thr Gly Cys Ala Ile
Ala Asn Ala Tyr 210 215 220
Thr Ala Leu Glu Ala Gly Ala Lys Leu Ile Asp Val Ser Val Leu Gly 225
230 235 240 Ile Gly Glu
Arg Asn Gly Ile Thr Pro Leu Gly Ala Leu Met Ala Arg 245
250 255 Met Ile Thr Ala Asp Arg Asp Tyr
Val Leu Ser Lys Tyr Lys Leu His 260 265
270 Lys Leu Arg Asp Leu Glu Asn Leu Val Ala Asp Ala Val
Gln Ile Asn 275 280 285
Ile Pro Phe Asn Asn Pro Ile Thr Gly Phe Cys Ala Phe Thr His Lys 290
295 300 Ala Gly Ile His
Ala Lys Ala Ile Leu Ala Asn Pro Ser Thr Tyr Glu 305 310
315 320 Ile Leu Asn Pro Asn Asp Phe Gly Leu
Thr Arg Tyr Ile His Phe Ala 325 330
335 Asn Arg Leu Thr Gly Trp Asn Ala Ile Lys Ser Arg Val Asp
Gln Leu 340 345 350
Asn Leu His Leu Thr Asp Asp Gln Val Lys Glu Val Thr Asn Lys Ile
355 360 365 Lys Lys Leu Gly
Asp Val Arg Gln Leu Asn Ile Asp Asp Val Asp Ser 370
375 380 Ile Ile Lys Asp Phe His Ala Glu
Gln Ser Thr Thr Asn Thr Pro Leu 385 390
395 400 Leu Lys Pro Val Glu Asp Asp Glu Gly Pro Glu Ile
Lys Lys Gln Lys 405 410
415 Val 391287DNACandida albicans 39atggttgacg gatataaaga agtatcagaa
tcatttgatc gttccaaaat ccaacataac 60ccttatggtc ctaatccagg tgattttctt
tcgaatgttg gcaattttca attgattgaa 120tcaactttga gagaaggtga acagtttgcc
aatgcatttt tcagcaccga aaccaaaatt 180gaaattgcta aagccttaga tgattttggg
gttgattata ttgaattgac ttcaccagtg 240gcatctgaac aatcaagaaa agattgtgaa
gccatttgta aattaggttt aaaagccaaa 300atattgactc acattagatg tcatatggat
gatgccagag ttgctgttga aactggggtc 360gatggagttg atgtggttat tggaacttcc
caatttttaa gacaatactc tcatggtaag 420gatatgaatt atattgcaca aagtgctatt
gaagtcattg aatttgtcaa atctaaaggt 480attgaaatcc gtttcagttc tgaagattct
tttagatcag atttggttga tttattaaac 540atttaccgta ctgttgataa aattggggtt
aacagagttg gtattgctga tactgttggt 600tgtgctaatc caagacaagt ttatgaattg
gtgagaacat tgaaatcagt agtcaagtgt 660gacattgaat gtcatttcca taatgatact
ggctgtgcca ttgccaatgc atacacagct 720ttggaaggtg gggccagatt gattgatgtt
tccgtattgg gtattggtga aagaaatggt 780attactccat tgggtgggtt aatggcgaga
atgattgctg ctgatcgtga atatgttttg 840tcaaaatata aagttcataa attgagagat
attgaaaatt tggttgctga ggcggttcaa 900gttaacattc cattcaataa tccgatcact
gggttctgtg ctttcactca taaagctggt 960atccatgcta aagctatctt ggccaatcca
tctacttatg aaattttgag tccaagtgat 1020ttcggtttaa ccagatatat tcactttgct
aatagattga ctggttggaa tgccatcaaa 1080tcaagagttg atcagttgaa cttgcattta
actgatgaac agtgtaaaga agtcactaac 1140aagattaaga aattgggtga tgtcagacaa
ttgaatatcg atgatgtgga ttcaatcatc 1200aaagatttcc atgctgatat gtcaacacca
cttttgaaat caaatggagc ggaagaagaa 1260ccagatgtaa aaaaacaaaa agtttaa
128740428PRTCandida albicans 40Met Val
Asp Gly Tyr Lys Glu Val Ser Glu Ser Phe Asp Arg Ser Lys 1 5
10 15 Ile Gln His Asn Pro Tyr Gly
Pro Asn Pro Gly Asp Phe Leu Ser Asn 20 25
30 Val Gly Asn Phe Gln Leu Ile Glu Ser Thr Leu Arg
Glu Gly Glu Gln 35 40 45
Phe Ala Asn Ala Phe Phe Ser Thr Glu Thr Lys Ile Glu Ile Ala Lys
50 55 60 Ala Leu Asp
Asp Phe Gly Val Asp Tyr Ile Glu Leu Thr Ser Pro Val 65
70 75 80 Ala Ser Glu Gln Ser Arg Lys
Asp Cys Glu Ala Ile Cys Lys Leu Gly 85
90 95 Leu Lys Ala Lys Ile Leu Thr His Ile Arg Cys
His Met Asp Asp Ala 100 105
110 Arg Val Ala Val Glu Thr Gly Val Asp Gly Val Asp Val Val Ile
Gly 115 120 125 Thr
Ser Gln Phe Leu Arg Gln Tyr Ser His Gly Lys Asp Met Asn Tyr 130
135 140 Ile Ala Gln Ser Ala Ile
Glu Val Ile Glu Phe Val Lys Ser Lys Gly 145 150
155 160 Ile Glu Ile Arg Phe Ser Ser Glu Asp Ser Phe
Arg Ser Asp Leu Val 165 170
175 Asp Leu Leu Asn Ile Tyr Arg Thr Val Asp Lys Ile Gly Val Asn Arg
180 185 190 Val Gly
Ile Ala Asp Thr Val Gly Cys Ala Asn Pro Arg Gln Val Tyr 195
200 205 Glu Leu Val Arg Thr Leu Lys
Ser Val Val Lys Cys Asp Ile Glu Cys 210 215
220 His Phe His Asn Asp Thr Gly Cys Ala Ile Ala Asn
Ala Tyr Thr Ala 225 230 235
240 Leu Glu Gly Gly Ala Arg Leu Ile Asp Val Ser Val Leu Gly Ile Gly
245 250 255 Glu Arg Asn
Gly Ile Thr Pro Leu Gly Gly Leu Met Ala Arg Met Ile 260
265 270 Ala Ala Asp Arg Glu Tyr Val Leu
Ser Lys Tyr Lys Val His Lys Leu 275 280
285 Arg Asp Ile Glu Asn Leu Val Ala Glu Ala Val Gln Val
Asn Ile Pro 290 295 300
Phe Asn Asn Pro Ile Thr Gly Phe Cys Ala Phe Thr His Lys Ala Gly 305
310 315 320 Ile His Ala Lys
Ala Ile Leu Ala Asn Pro Ser Thr Tyr Glu Ile Leu 325
330 335 Ser Pro Ser Asp Phe Gly Leu Thr Arg
Tyr Ile His Phe Ala Asn Arg 340 345
350 Leu Thr Gly Trp Asn Ala Ile Lys Ser Arg Val Asp Gln Leu
Asn Leu 355 360 365
His Leu Thr Asp Glu Gln Cys Lys Glu Val Thr Asn Lys Ile Lys Lys 370
375 380 Leu Gly Asp Val Arg
Gln Leu Asn Ile Asp Asp Val Asp Ser Ile Ile 385 390
395 400 Lys Asp Phe His Ala Asp Met Ser Thr Pro
Leu Leu Lys Ser Asn Gly 405 410
415 Ala Glu Glu Glu Pro Asp Val Lys Lys Gln Lys Val
420 425 41 1827DNACandida glabrata
41atgagagcta ccgttatcag actctcgagg gctgcgaagt caattccgcc cgtgaaattg
60gcgtataaga acatgttgaa agacccttcc atcaaataca aaccattctc cattgctcca
120aagcttactg acaggaaatg gccagacaat accattacca aggcaccaag gtggttgtct
180acagacttga gagacggtaa ccagtctctc ccggacccga tgtccattga gcagaagaaa
240gagtacttcc acaagctggt ggagattggc ttcaaagaga tagaagtcag ttttccatct
300gcctcgcaga ccgacttcga tttcacaagg tacgctgtgg agaatgctcc agatgatgtt
360accatacagt gtcttgtgca atccagagaa cacttgatca gaagaactgt ggagtcgttg
420actggtgcca agcgtgccac tatacatact tacttggcca ccagcgacat gttcagagag
480atagtcttca acatgtctaa ggaagacgct atcgccaaag ccgtcgaagc cactaaactg
540gtcagaagct tgaccaagga cgacccttct cagcaggcta cccgttggtc ctatgagttc
600tctccagaat gtttcagtga taccccagtc gaatttgccg ttgaaatctg tgaagcagta
660aaagctgcct gggaaccaac cgaggacaac cctatcatat ttaacctacc tgccacagtc
720gaggtcgcct ctccaaacat ctacgctgac caaatcgaat atttctgcac acacatcacc
780gaaagagaga aggtgtgtgt ctctacgcat acccacaacg accgtggctg cggtgttgcc
840gctaccgaac ttggtataat ggcaggcgct gatcgtgttg aaggttgtgt cttcggaaat
900ggtgaacgta ctggtaacgt tgacttggta accgtggcat tgaacatgta cacgcaaggt
960gtctctccta acttggactt ctccgacata aggtctgtaa tcgaggttgt tgaacgttgt
1020aacaaattgc ctgtcccagc cagagcacca tacggtggtg acttggtcgt atgtgcattc
1080tctggttctc accaggacgc catcaagaag ggtttctcgg ttcaacaaaa gaagcgtgac
1140caaggcgaca ttcaatggag aatcccatat ttgccattgg atccaaagga tatcggccgt
1200gactacgaag ctgtcatcag agtcaactct caatctggta agggtggtgc tgcttgggtt
1260gtcctaagag ccttgggcct agacatgcca agaaccatgc aaattgagtt ctccaccagt
1320gtacaagaac acgctgactc tctaggtaga gaactaaagg ccgaagagat tgtcaacttg
1380tttaaggaat cttacaacta caacaacgaa atcttccaac atatctcttt ggttgattac
1440aacgttgaga aattcggtgc tgagcgcaga attctaaatg gtcaagttga aatcaatggt
1500gaagttgtcg acatcaaggg taccggtaac ggtccaatct cttctttggt cgatgctttg
1560tccaacttat tgaacatcaa acttggtgtc agcaactata gtgaacactc tttgggttca
1620ggttcatcca ctcaagccgc ttctttcatc aacttaactt acagacgtga tgaagataat
1680gaaaaggctt accaatgggg tgtaggtgtg tctgaggatg ttggtgatgc ttctgtcaag
1740gcaatctttg ccactttgaa ttctgtaatt caaaaaggtg acattagtat cccaaagtct
1800aagaaggctg cctctggttc tgcttaa
182742608PRTCandida glabrata 42Met Arg Ala Thr Val Ile Arg Leu Ser Arg
Ala Ala Lys Ser Ile Pro 1 5 10
15 Pro Val Lys Leu Ala Tyr Lys Asn Met Leu Lys Asp Pro Ser Ile
Lys 20 25 30 Tyr
Lys Pro Phe Ser Ile Ala Pro Lys Leu Thr Asp Arg Lys Trp Pro 35
40 45 Asp Asn Thr Ile Thr Lys
Ala Pro Arg Trp Leu Ser Thr Asp Leu Arg 50 55
60 Asp Gly Asn Gln Ser Leu Pro Asp Pro Met Ser
Ile Glu Gln Lys Lys 65 70 75
80 Glu Tyr Phe His Lys Leu Val Glu Ile Gly Phe Lys Glu Ile Glu Val
85 90 95 Ser Phe
Pro Ser Ala Ser Gln Thr Asp Phe Asp Phe Thr Arg Tyr Ala 100
105 110 Val Glu Asn Ala Pro Asp Asp
Val Thr Ile Gln Cys Leu Val Gln Ser 115 120
125 Arg Glu His Leu Ile Arg Arg Thr Val Glu Ser Leu
Thr Gly Ala Lys 130 135 140
Arg Ala Thr Ile His Thr Tyr Leu Ala Thr Ser Asp Met Phe Arg Glu 145
150 155 160 Ile Val Phe
Asn Met Ser Lys Glu Asp Ala Ile Ala Lys Ala Val Glu 165
170 175 Ala Thr Lys Leu Val Arg Ser Leu
Thr Lys Asp Asp Pro Ser Gln Gln 180 185
190 Ala Thr Arg Trp Ser Tyr Glu Phe Ser Pro Glu Cys Phe
Ser Asp Thr 195 200 205
Pro Val Glu Phe Ala Val Glu Ile Cys Glu Ala Val Lys Ala Ala Trp 210
215 220 Glu Pro Thr Glu
Asp Asn Pro Ile Ile Phe Asn Leu Pro Ala Thr Val 225 230
235 240 Glu Val Ala Ser Pro Asn Ile Tyr Ala
Asp Gln Ile Glu Tyr Phe Cys 245 250
255 Thr His Ile Thr Glu Arg Glu Lys Val Cys Val Ser Thr His
Thr His 260 265 270
Asn Asp Arg Gly Cys Gly Val Ala Ala Thr Glu Leu Gly Ile Met Ala
275 280 285 Gly Ala Asp Arg
Val Glu Gly Cys Val Phe Gly Asn Gly Glu Arg Thr 290
295 300 Gly Asn Val Asp Leu Val Thr Val
Ala Leu Asn Met Tyr Thr Gln Gly 305 310
315 320 Val Ser Pro Asn Leu Asp Phe Ser Asp Ile Arg Ser
Val Ile Glu Val 325 330
335 Val Glu Arg Cys Asn Lys Leu Pro Val Pro Ala Arg Ala Pro Tyr Gly
340 345 350 Gly Asp Leu
Val Val Cys Ala Phe Ser Gly Ser His Gln Asp Ala Ile 355
360 365 Lys Lys Gly Phe Ser Val Gln Gln
Lys Lys Arg Asp Gln Gly Asp Ile 370 375
380 Gln Trp Arg Ile Pro Tyr Leu Pro Leu Asp Pro Lys Asp
Ile Gly Arg 385 390 395
400 Asp Tyr Glu Ala Val Ile Arg Val Asn Ser Gln Ser Gly Lys Gly Gly
405 410 415 Ala Ala Trp Val
Val Leu Arg Ala Leu Gly Leu Asp Met Pro Arg Thr 420
425 430 Met Gln Ile Glu Phe Ser Thr Ser Val
Gln Glu His Ala Asp Ser Leu 435 440
445 Gly Arg Glu Leu Lys Ala Glu Glu Ile Val Asn Leu Phe Lys
Glu Ser 450 455 460
Tyr Asn Tyr Asn Asn Glu Ile Phe Gln His Ile Ser Leu Val Asp Tyr 465
470 475 480 Asn Val Glu Lys Phe
Gly Ala Glu Arg Arg Ile Leu Asn Gly Gln Val 485
490 495 Glu Ile Asn Gly Glu Val Val Asp Ile Lys
Gly Thr Gly Asn Gly Pro 500 505
510 Ile Ser Ser Leu Val Asp Ala Leu Ser Asn Leu Leu Asn Ile Lys
Leu 515 520 525 Gly
Val Ser Asn Tyr Ser Glu His Ser Leu Gly Ser Gly Ser Ser Thr 530
535 540 Gln Ala Ala Ser Phe Ile
Asn Leu Thr Tyr Arg Arg Asp Glu Asp Asn 545 550
555 560 Glu Lys Ala Tyr Gln Trp Gly Val Gly Val Ser
Glu Asp Val Gly Asp 565 570
575 Ala Ser Val Lys Ala Ile Phe Ala Thr Leu Asn Ser Val Ile Gln Lys
580 585 590 Gly Asp
Ile Ser Ile Pro Lys Ser Lys Lys Ala Ala Ser Gly Ser Ala 595
600 605 43 1818DNACandida
glabrata 43atgagacaaa caattccaaa ttttgcagag catgtctctc gtgcagccaa
gacaattgct 60ccagtcaaat tgggtttcaa gaatatgctt gctaatccaa gtgtcaaata
tagaccattt 120caaggcccaa aattgacaaa tagacaatgg cctaacaaga caattaagag
agctccaaga 180tggctttcta ccgatttgag agatggtaac caatctctcc cggaccctat
gtcagtagag 240caaaagaaag aatactttca caaacttgtt gaaatcgggt ttaaagagat
agaagtcagt 300tttccgtcag catcgcaaac cgatttcgat ttcacaagat acgctgtaga
aaacgcacca 360gacgatgttt ctatccagtg tcttgtccaa tctagggagc atctgatcaa
gaggacagtt 420gaagcattga ccggtgctaa gcgtgctacc atacatacat acttggccac
aagtgatatg 480ttccgtgaga ttgttttcaa tatgtctcaa gaagaagcca ttgccaaagc
tgtagaagca 540accaagctag tacggaaatt gaccaaggat gatccatctc aaaaagcaac
taggtggtct 600tacgaatttt ctccagaatg ttttagtgat acaccagtag aatttgccgt
tgaaatctgt 660gaggctgtga aagctgcatg ggaaccaacg gttgataatc ctattatctt
taacttacct 720gcaaccgttg aagtagcaac tccaaatgta tacgctgatc agatcgaata
cttctctact 780catattagcg aacgtgaaaa ggtttgtatc tccacccatg ctcacaatga
ccgtggctgt 840ggcgttgctg ctacagagtt gggtatcttg gctggtgctg atcgagttga
aggctgtata 900ttcgggaatg gtgaacgtac aggtaatgtc gacctggtaa ccgtcgcctt
aaacatgtat 960acccagggtg tttctcccgg tcttgacttt tcagacatga gaagtgttat
cgagatcgtt 1020gaacgttgta acaagattcc agtaccagct agagctccat atggtggtga
ccttgttgtt 1080tgcgcctttt caggctctca ccaagatgct attaaaaaag gatttgcttt
acaacaaaag 1140aagcgtgctc aaggtgaaac tttatggagg attccatatt tgccattaga
tccaaaggac 1200atcggccgtg actatgaagc ggttatcagg gtcaactcac aatctggtaa
gggtggtgct 1260gcttgggtta ttttaaggtc tttgggtcta gacaccccaa gaaacatgca
aatgcaattc 1320tctaccattg tgcaaaatga agctgacaca agaggcaagg aattatctgc
agaggagatt 1380actgcattat tcaagtctac ctataattac aacaacgaaa cccatcaata
cgtatctttg 1440ctcgactatg atgtgaagaa gattgacaac gaccgtagaa tcctaacagg
gcaagttgaa 1500attaacgaca agatcattcc aattaagggt attggtaacg gtcctatttc
ttctttagta 1560gatgccctat caaacttatt caacgtcaaa tttggtgttg aaaactatac
agaacatgct 1620ttaggttccg gttccaaaac ccaagccgcc tctttcattc acatctctta
cagagatgct 1680gctaccaatg aaaaggagta cagttggggt gtcggtgtct ctgaagatgt
tggtgaagca 1740tctgttaggg ccattttctc aaccattaac agcattatcc attcaggtga
agtcactctt 1800cctactgaaa acaattag
181844605PRTCandida glabrata 44Met Arg Gln Thr Ile Pro Asn Phe
Ala Glu His Val Ser Arg Ala Ala 1 5 10
15 Lys Thr Ile Ala Pro Val Lys Leu Gly Phe Lys Asn Met
Leu Ala Asn 20 25 30
Pro Ser Val Lys Tyr Arg Pro Phe Gln Gly Pro Lys Leu Thr Asn Arg
35 40 45 Gln Trp Pro Asn
Lys Thr Ile Lys Arg Ala Pro Arg Trp Leu Ser Thr 50
55 60 Asp Leu Arg Asp Gly Asn Gln Ser
Leu Pro Asp Pro Met Ser Val Glu 65 70
75 80 Gln Lys Lys Glu Tyr Phe His Lys Leu Val Glu Ile
Gly Phe Lys Glu 85 90
95 Ile Glu Val Ser Phe Pro Ser Ala Ser Gln Thr Asp Phe Asp Phe Thr
100 105 110 Arg Tyr Ala
Val Glu Asn Ala Pro Asp Asp Val Ser Ile Gln Cys Leu 115
120 125 Val Gln Ser Arg Glu His Leu Ile
Lys Arg Thr Val Glu Ala Leu Thr 130 135
140 Gly Ala Lys Arg Ala Thr Ile His Thr Tyr Leu Ala Thr
Ser Asp Met 145 150 155
160 Phe Arg Glu Ile Val Phe Asn Met Ser Gln Glu Glu Ala Ile Ala Lys
165 170 175 Ala Val Glu Ala
Thr Lys Leu Val Arg Lys Leu Thr Lys Asp Asp Pro 180
185 190 Ser Gln Lys Ala Thr Arg Trp Ser Tyr
Glu Phe Ser Pro Glu Cys Phe 195 200
205 Ser Asp Thr Pro Val Glu Phe Ala Val Glu Ile Cys Glu Ala
Val Lys 210 215 220
Ala Ala Trp Glu Pro Thr Val Asp Asn Pro Ile Ile Phe Asn Leu Pro 225
230 235 240 Ala Thr Val Glu Val
Ala Thr Pro Asn Val Tyr Ala Asp Gln Ile Glu 245
250 255 Tyr Phe Ser Thr His Ile Ser Glu Arg Glu
Lys Val Cys Ile Ser Thr 260 265
270 His Ala His Asn Asp Arg Gly Cys Gly Val Ala Ala Thr Glu Leu
Gly 275 280 285 Ile
Leu Ala Gly Ala Asp Arg Val Glu Gly Cys Ile Phe Gly Asn Gly 290
295 300 Glu Arg Thr Gly Asn Val
Asp Leu Val Thr Val Ala Leu Asn Met Tyr 305 310
315 320 Thr Gln Gly Val Ser Pro Gly Leu Asp Phe Ser
Asp Met Arg Ser Val 325 330
335 Ile Glu Ile Val Glu Arg Cys Asn Lys Ile Pro Val Pro Ala Arg Ala
340 345 350 Pro Tyr
Gly Gly Asp Leu Val Val Cys Ala Phe Ser Gly Ser His Gln 355
360 365 Asp Ala Ile Lys Lys Gly Phe
Ala Leu Gln Gln Lys Lys Arg Ala Gln 370 375
380 Gly Glu Thr Leu Trp Arg Ile Pro Tyr Leu Pro Leu
Asp Pro Lys Asp 385 390 395
400 Ile Gly Arg Asp Tyr Glu Ala Val Ile Arg Val Asn Ser Gln Ser Gly
405 410 415 Lys Gly Gly
Ala Ala Trp Val Ile Leu Arg Ser Leu Gly Leu Asp Thr 420
425 430 Pro Arg Asn Met Gln Met Gln Phe
Ser Thr Ile Val Gln Asn Glu Ala 435 440
445 Asp Thr Arg Gly Lys Glu Leu Ser Ala Glu Glu Ile Thr
Ala Leu Phe 450 455 460
Lys Ser Thr Tyr Asn Tyr Asn Asn Glu Thr His Gln Tyr Val Ser Leu 465
470 475 480 Leu Asp Tyr Asp
Val Lys Lys Ile Asp Asn Asp Arg Arg Ile Leu Thr 485
490 495 Gly Gln Val Glu Ile Asn Asp Lys Ile
Ile Pro Ile Lys Gly Ile Gly 500 505
510 Asn Gly Pro Ile Ser Ser Leu Val Asp Ala Leu Ser Asn Leu
Phe Asn 515 520 525
Val Lys Phe Gly Val Glu Asn Tyr Thr Glu His Ala Leu Gly Ser Gly 530
535 540 Ser Lys Thr Gln Ala
Ala Ser Phe Ile His Ile Ser Tyr Arg Asp Ala 545 550
555 560 Ala Thr Asn Glu Lys Glu Tyr Ser Trp Gly
Val Gly Val Ser Glu Asp 565 570
575 Val Gly Glu Ala Ser Val Arg Ala Ile Phe Ser Thr Ile Asn Ser
Ile 580 585 590 Ile
His Ser Gly Glu Val Thr Leu Pro Thr Glu Asn Asn 595
600 605 45 1752DNAKluyveromyces lactis 45atgccattct
acaaagatcc ttcagtgaag tataaaccat tcgttagcaa cgtcaaatta 60caggacagga
aatggcctag taaaaccctt aataaggctc caagatggtt agctaccgat 120ttaagagatg
ggaatcagtc tttacctgac ccgatgaatt tggaagagaa gaaactgatg 180ctcgataagt
tatgcgaatt gggtttcaaa gagattgaag ttgctttccc tagtgcttct 240aatatcgatt
tccaattcac tcaatatgca gtgaaaaacg taccagaaga cgtttccatt 300caagttcttt
ctccatgtcg tgaacccttg atcgaacgta ccgttgaatc tttggtcggt 360gccaagagag
ccattgtaca tatctatctc gcgacatcac catgtttcag agaaatcgtt 420ttcaacaata
tgtctcatga agaaagtatt gaaaaggctg tgaaatgtgc caaacttgtt 480aggtccttga
caaaagacca tccggataga caagataccc attggtcatt tgagttttct 540ccagaaacgt
tcagcgatag tgaaccggat ttcgttctag agatttgtga agctgttaag 600gctgcttggg
gacccactga agataatcca atcattttca atttgccagc taccgtcgag 660atggctacac
caaacgtgta cgctgaccaa atcgaatatt tcgctcaaag tatctccgaa 720cgtgagaaag
tatgtatctc tctccatcca cataacgatc gtgggtgtgc tgtggcagct 780gcagaattag
ctcaaatggc tggtgcagat cgtgtcgagg gatgtctctt cggtaacggt 840gaacgtaccg
gtaacgttga tttggttacc ttggcattaa acctctacac acagggtgta 900tctccaaacc
tcgatttctc cgatatggct tctattattg aagtcgttga gaaatgtaat 960aagattcccg
tgcatgctag agcaccctac ggaggacaac ttgtcgtttg tgcattcagt 1020ggttctcatc
aagatgccat caaaaagggt ttcgaaaaat acgacaacaa ggttaaggct 1080ttacaagaaa
aagagggtcc agatgcagtg gtaccttgga aaatgccata tctccccttg 1140gatcctcagg
atattggaag aacgtatgag gctatcatca gagtcaactc gcaatcaggt 1200aaaggtggtt
cttcttgggt tatcctaaag aacttggagc tagatttacc aagagatcta 1260caaattgcat
actctaagat cgttcaaaat gaaactgaga tagtcggtag agagttgaag 1320agcgatgaac
taatctcttt attcgagaaa tcgtatttcg ttggatctca ttcaactact 1380ggtaaattca
agtttatcga ctataaatat gacaaatctc cggagaattt cactctttcg 1440gtgcagctat
cagatggaac tactcaatgg gatttggaag gtactggtaa cggtccaatc 1500tcttctttca
tcgatgctgt gaataaaaac ttcaaaacta atcttgatgt gaaaaactat 1560catgagcatt
ccttgggtaa gagttccgat tcgagagctg ctacttatat ctctgtctct 1620catgaaggat
ttgttcaatg gggtgttggt attcatgagg atactactct ggcttcattc 1680ttggcgttgt
tatcttgtat aaacggtctt gatagggcaa agaacttcac tgtcaattca 1740gctgccaatt
ga
175246583PRTKluyveromyces lactis 46Met Pro Phe Tyr Lys Asp Pro Ser Val
Lys Tyr Lys Pro Phe Val Ser 1 5 10
15 Asn Val Lys Leu Gln Asp Arg Lys Trp Pro Ser Lys Thr Leu
Asn Lys 20 25 30
Ala Pro Arg Trp Leu Ala Thr Asp Leu Arg Asp Gly Asn Gln Ser Leu
35 40 45 Pro Asp Pro Met
Asn Leu Glu Glu Lys Lys Leu Met Leu Asp Lys Leu 50
55 60 Cys Glu Leu Gly Phe Lys Glu Ile
Glu Val Ala Phe Pro Ser Ala Ser 65 70
75 80 Asn Ile Asp Phe Gln Phe Thr Gln Tyr Ala Val Lys
Asn Val Pro Glu 85 90
95 Asp Val Ser Ile Gln Val Leu Ser Pro Cys Arg Glu Pro Leu Ile Glu
100 105 110 Arg Thr Val
Glu Ser Leu Val Gly Ala Lys Arg Ala Ile Val His Ile 115
120 125 Tyr Leu Ala Thr Ser Pro Cys Phe
Arg Glu Ile Val Phe Asn Asn Met 130 135
140 Ser His Glu Glu Ser Ile Glu Lys Ala Val Lys Cys Ala
Lys Leu Val 145 150 155
160 Arg Ser Leu Thr Lys Asp His Pro Asp Arg Gln Asp Thr His Trp Ser
165 170 175 Phe Glu Phe Ser
Pro Glu Thr Phe Ser Asp Ser Glu Pro Asp Phe Val 180
185 190 Leu Glu Ile Cys Glu Ala Val Lys Ala
Ala Trp Gly Pro Thr Glu Asp 195 200
205 Asn Pro Ile Ile Phe Asn Leu Pro Ala Thr Val Glu Met Ala
Thr Pro 210 215 220
Asn Val Tyr Ala Asp Gln Ile Glu Tyr Phe Ala Gln Ser Ile Ser Glu 225
230 235 240 Arg Glu Lys Val Cys
Ile Ser Leu His Pro His Asn Asp Arg Gly Cys 245
250 255 Ala Val Ala Ala Ala Glu Leu Ala Gln Met
Ala Gly Ala Asp Arg Val 260 265
270 Glu Gly Cys Leu Phe Gly Asn Gly Glu Arg Thr Gly Asn Val Asp
Leu 275 280 285 Val
Thr Leu Ala Leu Asn Leu Tyr Thr Gln Gly Val Ser Pro Asn Leu 290
295 300 Asp Phe Ser Asp Met Ala
Ser Ile Ile Glu Val Val Glu Lys Cys Asn 305 310
315 320 Lys Ile Pro Val His Ala Arg Ala Pro Tyr Gly
Gly Gln Leu Val Val 325 330
335 Cys Ala Phe Ser Gly Ser His Gln Asp Ala Ile Lys Lys Gly Phe Glu
340 345 350 Lys Tyr
Asp Asn Lys Val Lys Ala Leu Gln Glu Lys Glu Gly Pro Asp 355
360 365 Ala Val Val Pro Trp Lys Met
Pro Tyr Leu Pro Leu Asp Pro Gln Asp 370 375
380 Ile Gly Arg Thr Tyr Glu Ala Ile Ile Arg Val Asn
Ser Gln Ser Gly 385 390 395
400 Lys Gly Gly Ser Ser Trp Val Ile Leu Lys Asn Leu Glu Leu Asp Leu
405 410 415 Pro Arg Asp
Leu Gln Ile Ala Tyr Ser Lys Ile Val Gln Asn Glu Thr 420
425 430 Glu Ile Val Gly Arg Glu Leu Lys
Ser Asp Glu Leu Ile Ser Leu Phe 435 440
445 Glu Lys Ser Tyr Phe Val Gly Ser His Ser Thr Thr Gly
Lys Phe Lys 450 455 460
Phe Ile Asp Tyr Lys Tyr Asp Lys Ser Pro Glu Asn Phe Thr Leu Ser 465
470 475 480 Val Gln Leu Ser
Asp Gly Thr Thr Gln Trp Asp Leu Glu Gly Thr Gly 485
490 495 Asn Gly Pro Ile Ser Ser Phe Ile Asp
Ala Val Asn Lys Asn Phe Lys 500 505
510 Thr Asn Leu Asp Val Lys Asn Tyr His Glu His Ser Leu Gly
Lys Ser 515 520 525
Ser Asp Ser Arg Ala Ala Thr Tyr Ile Ser Val Ser His Glu Gly Phe 530
535 540 Val Gln Trp Gly Val
Gly Ile His Glu Asp Thr Thr Leu Ala Ser Phe 545 550
555 560 Leu Ala Leu Leu Ser Cys Ile Asn Gly Leu
Asp Arg Ala Lys Asn Phe 565 570
575 Thr Val Asn Ser Ala Ala Asn 580 47
1830DNAKluyveromyces lactis 47atgatattca ggaacaccgt tgtgcgttta
gcacaggctg ggaaaaaagc tattcctcca 60gtgaaactag cgtacaagaa tatgctaaaa
gatccatcga cgaaatatag accataccca 120cagatcaact tggaaaatag acaatggcct
tcgaagacca tcaccaaggc tcctaggtgg 180ctttctaccg atctaagaga cgggaatcaa
tctttaccag atcctatgtc tgtcgagcag 240aagaaggaat atttccataa gttgattgag
attggtttca aagaaattga ggtctcattc 300ccatctgcgt cgcaaacaga tttcgacttc
acaagatacg ctgttgaaaa cgccccagaa 360gatgtttcca ttcaatgtct tgttcaatcg
agagaacatt tgattagaag aacagttgaa 420gctttgcatg gtgctaagaa agccaccatc
catacgtatt tggccacctc cgacatgttc 480cgtgacattg tgttcaacat gtcccaagaa
gaagctattg ctaaagctgt ggaagccacc 540aagttagtta ggaaattgac caaggatgat
ccttcgcaaa gtgctacaca atggacttac 600cagttctctc cagaatgttt cagtgataca
cctgtagaat ttgctgttga gatctgtgaa 660gccgtaaagg ctgcttggga accaactgag
gaaaacccaa tcattttcaa cctacctgct 720accgtcgaag tcgctactcc aaacatttac
gctgatcaaa ttgaatactt ttcaactcac 780atatctgaac gtgaaaaggt ctgtatctcc
acacatgcgc acaacgaccg tggctgtggt 840gttgctgctt ctgaactagg tattttggct
ggtgctgacc gtgtcgaagg ttgtttattc 900ggtaatggtg aacgtactgg taacgttgac
ttggttactg tcgcattgaa catgtacact 960caaggtgttt ctccagaatt agacttatct
gatattaact cagtcattga agtagtggaa 1020agatgtaaca agattgcagt ttcaccaaga
gccccatatg gtggtgactt ggtcgtttgt 1080gctttcagtg gttctcatca agatgctatc
aaaaagggtt tcaatcttca agaaaagaga 1140cgtagtcaag gtgatactct atggaaaatt
ccatacttgc cattggatcc aaaggatatc 1200ggtagagact acgaagctgt catccgtgtc
aactctcaat ctggtaaggg gggtgccgct 1260tgggttgtct tgagatcttt gggcctagat
ttgccaagaa acttgcaaat tgaattttcc 1320actcaagtgc aagaaaaggc tgatgctcta
ggtaaggaac taaaggcaaa cgaaattgtc 1380agcaccttca agtcgttata caacctcgat
ggaagcgcct ccaacatttc tttgttagaa 1440tacaatgttt ctaaagtaca gggtgatcag
aagagttttg ttggtcaagt ccagatcgac 1500aacgaagtcg tcggcattga aggtctcgga
aacggtccaa tttcctctct aatcgatgcg 1560ttgtcaaatt tgctcggtgt taaacttggt
gttgccaact acaccgaaca ttccttagga 1620tctggttctt caacaaaggc tgcttcttac
gtgcatattg cttacagaag agaaattgac 1680aacgaaaagg cctaccaatg gggtattggt
atgtctgaag atgttggaga ggcttctgcc 1740aaagccatcc tttctgctgt taataacttg
atcaaaaagg gcgaactaac aataccagct 1800catcgtgact cagcctcagc atctgcatag
183048609PRTKluyveromyces lactis 48Met
Ile Phe Arg Asn Thr Val Val Arg Leu Ala Gln Ala Gly Lys Lys 1
5 10 15 Ala Ile Pro Pro Val Lys
Leu Ala Tyr Lys Asn Met Leu Lys Asp Pro 20
25 30 Ser Thr Lys Tyr Arg Pro Tyr Pro Gln Ile
Asn Leu Glu Asn Arg Gln 35 40
45 Trp Pro Ser Lys Thr Ile Thr Lys Ala Pro Arg Trp Leu Ser
Thr Asp 50 55 60
Leu Arg Asp Gly Asn Gln Ser Leu Pro Asp Pro Met Ser Val Glu Gln 65
70 75 80 Lys Lys Glu Tyr Phe
His Lys Leu Ile Glu Ile Gly Phe Lys Glu Ile 85
90 95 Glu Val Ser Phe Pro Ser Ala Ser Gln Thr
Asp Phe Asp Phe Thr Arg 100 105
110 Tyr Ala Val Glu Asn Ala Pro Glu Asp Val Ser Ile Gln Cys Leu
Val 115 120 125 Gln
Ser Arg Glu His Leu Ile Arg Arg Thr Val Glu Ala Leu His Gly 130
135 140 Ala Lys Lys Ala Thr Ile
His Thr Tyr Leu Ala Thr Ser Asp Met Phe 145 150
155 160 Arg Asp Ile Val Phe Asn Met Ser Gln Glu Glu
Ala Ile Ala Lys Ala 165 170
175 Val Glu Ala Thr Lys Leu Val Arg Lys Leu Thr Lys Asp Asp Pro Ser
180 185 190 Gln Ser
Ala Thr Gln Trp Thr Tyr Gln Phe Ser Pro Glu Cys Phe Ser 195
200 205 Asp Thr Pro Val Glu Phe Ala
Val Glu Ile Cys Glu Ala Val Lys Ala 210 215
220 Ala Trp Glu Pro Thr Glu Glu Asn Pro Ile Ile Phe
Asn Leu Pro Ala 225 230 235
240 Thr Val Glu Val Ala Thr Pro Asn Ile Tyr Ala Asp Gln Ile Glu Tyr
245 250 255 Phe Ser Thr
His Ile Ser Glu Arg Glu Lys Val Cys Ile Ser Thr His 260
265 270 Ala His Asn Asp Arg Gly Cys Gly
Val Ala Ala Ser Glu Leu Gly Ile 275 280
285 Leu Ala Gly Ala Asp Arg Val Glu Gly Cys Leu Phe Gly
Asn Gly Glu 290 295 300
Arg Thr Gly Asn Val Asp Leu Val Thr Val Ala Leu Asn Met Tyr Thr 305
310 315 320 Gln Gly Val Ser
Pro Glu Leu Asp Leu Ser Asp Ile Asn Ser Val Ile 325
330 335 Glu Val Val Glu Arg Cys Asn Lys Ile
Ala Val Ser Pro Arg Ala Pro 340 345
350 Tyr Gly Gly Asp Leu Val Val Cys Ala Phe Ser Gly Ser His
Gln Asp 355 360 365
Ala Ile Lys Lys Gly Phe Asn Leu Gln Glu Lys Arg Arg Ser Gln Gly 370
375 380 Asp Thr Leu Trp Lys
Ile Pro Tyr Leu Pro Leu Asp Pro Lys Asp Ile 385 390
395 400 Gly Arg Asp Tyr Glu Ala Val Ile Arg Val
Asn Ser Gln Ser Gly Lys 405 410
415 Gly Gly Ala Ala Trp Val Val Leu Arg Ser Leu Gly Leu Asp Leu
Pro 420 425 430 Arg
Asn Leu Gln Ile Glu Phe Ser Thr Gln Val Gln Glu Lys Ala Asp 435
440 445 Ala Leu Gly Lys Glu Leu
Lys Ala Asn Glu Ile Val Ser Thr Phe Lys 450 455
460 Ser Leu Tyr Asn Leu Asp Gly Ser Ala Ser Asn
Ile Ser Leu Leu Glu 465 470 475
480 Tyr Asn Val Ser Lys Val Gln Gly Asp Gln Lys Ser Phe Val Gly Gln
485 490 495 Val Gln
Ile Asp Asn Glu Val Val Gly Ile Glu Gly Leu Gly Asn Gly 500
505 510 Pro Ile Ser Ser Leu Ile Asp
Ala Leu Ser Asn Leu Leu Gly Val Lys 515 520
525 Leu Gly Val Ala Asn Tyr Thr Glu His Ser Leu Gly
Ser Gly Ser Ser 530 535 540
Thr Lys Ala Ala Ser Tyr Val His Ile Ala Tyr Arg Arg Glu Ile Asp 545
550 555 560 Asn Glu Lys
Ala Tyr Gln Trp Gly Ile Gly Met Ser Glu Asp Val Gly 565
570 575 Glu Ala Ser Ala Lys Ala Ile Leu
Ser Ala Val Asn Asn Leu Ile Lys 580 585
590 Lys Gly Glu Leu Thr Ile Pro Ala His Arg Asp Ser Ala
Ser Ala Ser 595 600 605
Ala 491341DNAYarrowia lipolytica 49atgtgcgcaa cagataacgc ccccgccgct
aacgctgctc ctgagaagcc ctccaacgtt 60ggagtcgagg tcggtcacac cggcgagcag
actaatcctt acggagccaa ccccgccgat 120ttcctttcta acgtgtccaa gttccagctc
atcgagtcca ctctgcgaga gggagagcag 180tttgcctctg ccttctttga caccgagacc
aagatcgaga ttgccaaggc tctggacgac 240tttggtgtcg actacatcga gctgacctcc
cccgcagcat cggagcagtc gcggtccgat 300tgcgaggcca tctgcaagct cggtcttaag
gccaagattc tcactcacat ccgatgccac 360atggacgacg caagactcgc tgtctccacc
ggtgtcgatg gtgtcgatgt cgtcattggt 420acctcccagt tcctgcgaca gtactcccac
ggcaaggaca tgaactacat tgcacagtcc 480gctgtcgagg tcattgagtt tgtcaagagc
cacggcattg agatccgatt ctcctccgag 540gattctttcc gatccgacct ggtcgatctc
ctcaacatct accgaactgt cgacaagatt 600ggtgtcaacc gagtcggtat tgctgacact
gttggatgcg ccaacccccg acaggtctac 660gagcttgtcc gaaccctcaa gtccgttgtc
tcgtgcgaca ttgagtgcca tttccacaac 720gacaccggct gtgccattgc caacgcctac
accgccctcg aggctggtgc caacctcatc 780gatgtctccg ttctcggtat cggtgagcga
aacggtatca cctctctcgg tggtctgatg 840gctcgaatga ttgctgctga ccgagactac
gttctctcca agtacaagct gcacaagctg 900cgagacctcg agaacctcgt cgccgacgcc
gtccaggtca acatcccctt caacaacccc 960atcaccggtt tctgcgcctt cacccacaag
gccggtatcc acgccaaggc cattctcgcc 1020aacccctcca cttacgagat tctcaacccc
gccgatttcg gtctgacccg atacatccac 1080tttgccaacc gtcttaccgg ctggaacgcc
atcaagtcgc gagttgacca gctcaacctg 1140cacctgaccg acgcccagtg caaggatgtc
actgccaaga tcaagaagct tggtgacgtt 1200cgatctctca acattgacga tgttgactcc
atcatccgag agttccacgc cgatgtcacc 1260agcactccca ccgttgctgc caccgaggga
cctgccgttg aggacgagcc cgccgccaag 1320aaggccaaga ctgaagagta a
134150446PRTYarrowia lipolytica 50Met
Cys Ala Thr Asp Asn Ala Pro Ala Ala Asn Ala Ala Pro Glu Lys 1
5 10 15 Pro Ser Asn Val Gly Val
Glu Val Gly His Thr Gly Glu Gln Thr Asn 20
25 30 Pro Tyr Gly Ala Asn Pro Ala Asp Phe Leu
Ser Asn Val Ser Lys Phe 35 40
45 Gln Leu Ile Glu Ser Thr Leu Arg Glu Gly Glu Gln Phe Ala
Ser Ala 50 55 60
Phe Phe Asp Thr Glu Thr Lys Ile Glu Ile Ala Lys Ala Leu Asp Asp 65
70 75 80 Phe Gly Val Asp Tyr
Ile Glu Leu Thr Ser Pro Ala Ala Ser Glu Gln 85
90 95 Ser Arg Ser Asp Cys Glu Ala Ile Cys Lys
Leu Gly Leu Lys Ala Lys 100 105
110 Ile Leu Thr His Ile Arg Cys His Met Asp Asp Ala Arg Leu Ala
Val 115 120 125 Ser
Thr Gly Val Asp Gly Val Asp Val Val Ile Gly Thr Ser Gln Phe 130
135 140 Leu Arg Gln Tyr Ser His
Gly Lys Asp Met Asn Tyr Ile Ala Gln Ser 145 150
155 160 Ala Val Glu Val Ile Glu Phe Val Lys Ser His
Gly Ile Glu Ile Arg 165 170
175 Phe Ser Ser Glu Asp Ser Phe Arg Ser Asp Leu Val Asp Leu Leu Asn
180 185 190 Ile Tyr
Arg Thr Val Asp Lys Ile Gly Val Asn Arg Val Gly Ile Ala 195
200 205 Asp Thr Val Gly Cys Ala Asn
Pro Arg Gln Val Tyr Glu Leu Val Arg 210 215
220 Thr Leu Lys Ser Val Val Ser Cys Asp Ile Glu Cys
His Phe His Asn 225 230 235
240 Asp Thr Gly Cys Ala Ile Ala Asn Ala Tyr Thr Ala Leu Glu Ala Gly
245 250 255 Ala Asn Leu
Ile Asp Val Ser Val Leu Gly Ile Gly Glu Arg Asn Gly 260
265 270 Ile Thr Ser Leu Gly Gly Leu Met
Ala Arg Met Ile Ala Ala Asp Arg 275 280
285 Asp Tyr Val Leu Ser Lys Tyr Lys Leu His Lys Leu Arg
Asp Leu Glu 290 295 300
Asn Leu Val Ala Asp Ala Val Gln Val Asn Ile Pro Phe Asn Asn Pro 305
310 315 320 Ile Thr Gly Phe
Cys Ala Phe Thr His Lys Ala Gly Ile His Ala Lys 325
330 335 Ala Ile Leu Ala Asn Pro Ser Thr Tyr
Glu Ile Leu Asn Pro Ala Asp 340 345
350 Phe Gly Leu Thr Arg Tyr Ile His Phe Ala Asn Arg Leu Thr
Gly Trp 355 360 365
Asn Ala Ile Lys Ser Arg Val Asp Gln Leu Asn Leu His Leu Thr Asp 370
375 380 Ala Gln Cys Lys Asp
Val Thr Ala Lys Ile Lys Lys Leu Gly Asp Val 385 390
395 400 Arg Ser Leu Asn Ile Asp Asp Val Asp Ser
Ile Ile Arg Glu Phe His 405 410
415 Ala Asp Val Thr Ser Thr Pro Thr Val Ala Ala Thr Glu Gly Pro
Ala 420 425 430 Val
Glu Asp Glu Pro Ala Ala Lys Lys Ala Lys Thr Glu Glu 435
440 445 51 1728DNAYarrowia lipolytica
51atgccctacc tggccgatcc ctccaccaaa tacaagccgt tccccccgat caatctgccc
60aaccggcagt ggccgtcgaa aacgctgcag aagcccccgc ggtggctgtc gacggacctg
120cgggacggca accagtcgct gccggatccc atgaccatgg cggagaagaa ggagtacttc
180cagaagattg tcgacattgg ctacaaggag atcgaggtgg cgttcccgtc cgcctcgcag
240gtggactttg acttcacccg ctttgcctgc gacaccgccc ccgaagacgt gtggatccag
300gtgctggctc cgtgccgaga ggatctcatc acccgaaccg tcgagtccgt caagggcgcc
360aacaaggcca tcatccacat ctacctcgcc acctccaagt gcttccggga cattgtcttc
420aaccattcgc gagaagaggc cctggccaag gccgtggcat gcgccaagca cgtgcgagcc
480ctgaccaagg actcggacga ccccgagtgc aaaaagacca cctggggttt tgagttctcc
540cccgagacct tctccgacac cgacgtggac tacgccattg aggtctgtga ggccgtgaag
600gccgcctggg gcccctccga ggagaacccc atcattttca acctccccgc caccgtcgaa
660atggccaccc ccaacatcta cgccgaccag attgagtact ttgccaccaa catttccgag
720cgggagaaga tttgcatttc tctgcacccc cacaacgacc gaggttgtgc cgtggctgct
780gccgagctgg gccagatggc cggagccgac cgagtcgagg gctgtctgtt tggcaacggc
840gagcgaaccg gaaacgtcga cctcgtcact ctgggtctga atttgtacac ccagggcgtg
900catcccaaga ttgacttctc cgacatcacc tcgatcatcg acattgtgga gcgatgcaac
960aagatccccg tgcacccccg agctccctac ggcggccagc tggtggtgtg tgccttctcc
1020ggctctcacc aggacgccat caagaagggc tttgctcgaa tcgaagacgt caaggatgag
1080gtggccgagg gcaagcgaca gtggcagatc ccctacctgc ctcttgaccc caaggacatt
1140ggccgaacct acgaggcagt cattcgagtc aattcgcagt ccggcaaggg aggagccgcc
1200tggatcattc tgcgatctct ggagctcgat ctgccccgag gcctgcaggt tgccttctcc
1260aaggtggtcc agaaggaggc cgaggtggtt ggacaggagc tgtctgccca gcagttggtg
1320gatctctttg agcgagagta cggcgtgttt gaggagcagc agggcaagta ccagctggac
1380gactttgagg tgaccaacaa gtccaaggag gagcgagagc tgaccggagc tctgaccgtc
1440gagggcaagc gagtcgagct caagggtacc ggtaacggtc ccatttcgtc cttcctggat
1500gccatcaaga acgcctttgg ctacaacctc gaggttctca actaccacga gcactccatt
1560ggtaagggtt ccaagaccaa ggctgctact tacattgagc tggcctatga ggaggacggc
1620aagacttcca agcgatgggg tgttggtatt gacgaggatg tttcccaggc ttctattcat
1680gctattctgt ctgccatgaa cgccattagc gagtcctaca agaaataa
172852575PRTYarrowia lipolytica 52Met Pro Tyr Leu Ala Asp Pro Ser Thr Lys
Tyr Lys Pro Phe Pro Pro 1 5 10
15 Ile Asn Leu Pro Asn Arg Gln Trp Pro Ser Lys Thr Leu Gln Lys
Pro 20 25 30 Pro
Arg Trp Leu Ser Thr Asp Leu Arg Asp Gly Asn Gln Ser Leu Pro 35
40 45 Asp Pro Met Thr Met Ala
Glu Lys Lys Glu Tyr Phe Gln Lys Ile Val 50 55
60 Asp Ile Gly Tyr Lys Glu Ile Glu Val Ala Phe
Pro Ser Ala Ser Gln 65 70 75
80 Val Asp Phe Asp Phe Thr Arg Phe Ala Cys Asp Thr Ala Pro Glu Asp
85 90 95 Val Trp
Ile Gln Val Leu Ala Pro Cys Arg Glu Asp Leu Ile Thr Arg 100
105 110 Thr Val Glu Ser Val Lys Gly
Ala Asn Lys Ala Ile Ile His Ile Tyr 115 120
125 Leu Ala Thr Ser Lys Cys Phe Arg Asp Ile Val Phe
Asn His Ser Arg 130 135 140
Glu Glu Ala Leu Ala Lys Ala Val Ala Cys Ala Lys His Val Arg Ala 145
150 155 160 Leu Thr Lys
Asp Ser Asp Asp Pro Glu Cys Lys Lys Thr Thr Trp Gly 165
170 175 Phe Glu Phe Ser Pro Glu Thr Phe
Ser Asp Thr Asp Val Asp Tyr Ala 180 185
190 Ile Glu Val Cys Glu Ala Val Lys Ala Ala Trp Gly Pro
Ser Glu Glu 195 200 205
Asn Pro Ile Ile Phe Asn Leu Pro Ala Thr Val Glu Met Ala Thr Pro 210
215 220 Asn Ile Tyr Ala
Asp Gln Ile Glu Tyr Phe Ala Thr Asn Ile Ser Glu 225 230
235 240 Arg Glu Lys Ile Cys Ile Ser Leu His
Pro His Asn Asp Arg Gly Cys 245 250
255 Ala Val Ala Ala Ala Glu Leu Gly Gln Met Ala Gly Ala Asp
Arg Val 260 265 270
Glu Gly Cys Leu Phe Gly Asn Gly Glu Arg Thr Gly Asn Val Asp Leu
275 280 285 Val Thr Leu Gly
Leu Asn Leu Tyr Thr Gln Gly Val His Pro Lys Ile 290
295 300 Asp Phe Ser Asp Ile Thr Ser Ile
Ile Asp Ile Val Glu Arg Cys Asn 305 310
315 320 Lys Ile Pro Val His Pro Arg Ala Pro Tyr Gly Gly
Gln Leu Val Val 325 330
335 Cys Ala Phe Ser Gly Ser His Gln Asp Ala Ile Lys Lys Gly Phe Ala
340 345 350 Arg Ile Glu
Asp Val Lys Asp Glu Val Ala Glu Gly Lys Arg Gln Trp 355
360 365 Gln Ile Pro Tyr Leu Pro Leu Asp
Pro Lys Asp Ile Gly Arg Thr Tyr 370 375
380 Glu Ala Val Ile Arg Val Asn Ser Gln Ser Gly Lys Gly
Gly Ala Ala 385 390 395
400 Trp Ile Ile Leu Arg Ser Leu Glu Leu Asp Leu Pro Arg Gly Leu Gln
405 410 415 Val Ala Phe Ser
Lys Val Val Gln Lys Glu Ala Glu Val Val Gly Gln 420
425 430 Glu Leu Ser Ala Gln Gln Leu Val Asp
Leu Phe Glu Arg Glu Tyr Gly 435 440
445 Val Phe Glu Glu Gln Gln Gly Lys Tyr Gln Leu Asp Asp Phe
Glu Val 450 455 460
Thr Asn Lys Ser Lys Glu Glu Arg Glu Leu Thr Gly Ala Leu Thr Val 465
470 475 480 Glu Gly Lys Arg Val
Glu Leu Lys Gly Thr Gly Asn Gly Pro Ile Ser 485
490 495 Ser Phe Leu Asp Ala Ile Lys Asn Ala Phe
Gly Tyr Asn Leu Glu Val 500 505
510 Leu Asn Tyr His Glu His Ser Ile Gly Lys Gly Ser Lys Thr Lys
Ala 515 520 525 Ala
Thr Tyr Ile Glu Leu Ala Tyr Glu Glu Asp Gly Lys Thr Ser Lys 530
535 540 Arg Trp Gly Val Gly Ile
Asp Glu Asp Val Ser Gln Ala Ser Ile His 545 550
555 560 Ala Ile Leu Ser Ala Met Asn Ala Ile Ser Glu
Ser Tyr Lys Lys 565 570
575 531716DNAPichia stipitis 53atgttaaagg atccttccac caaatatgct
gcctttaaag gagtcaagtt ggacaagaga 60acctggccct caaagtctat caccaaggct
cctaggtggt tatctactga tttaagagat 120ggtaaccaag cgttgcctga tcccatgtct
gtcgaagaga agaaggagta ttttcacaag 180ctcttggaga tcggattcaa agaaatcgag
gtatctttcc cttctgcatc tcaaacagat 240tttgacttca ccagatatgc tgtggagaac
gcaccagatg atgtttcgat ccaagttttg 300actcagtcta gagaaccttt gatcagaaga
actgttgaat ccgtaaaggg tgctaagaag 360gctaccatac atacatactt ggctacttct
gacgttttcc gtgatgttgt tttcaacatg 420tcacaagaag atgcaattgc caaagccatt
gaaactacca agttggtcaa gtctttgaca 480aaggacgatc cagaaatgca ggaaaccgag
tggaccttgg aattctctcc tgaatgcttc 540tcagatactc ctaccgaatt tgctgtgcaa
atttgtgaag cagtcaagaa cgtctgggag 600ccaactgtag agaatcctat cattttcaac
ttgccagcta ccgttgaagt tgcttctcca 660aacgtctacg ctgaccagat cgaatacttt
gctacccaca tttccgaacg tgaaaaggtg 720tgtatttctc ttcatgctca caatgaccgt
ggctgcggtg ttgctgcctc ggaattaggt 780ttattggctg gcggagacag agtcgaaggt
tgtttgtttg gaaacggtga aagaaccggt 840aacgtagact tgatcactgt tgctctcaac
atgtacacca atggagttgc accggagttg 900gacttttcag aaatcgaaaa gctcatcgag
gtcagtgaaa gatgtaacaa aatcccagtt 960cacccaagag ctccatactc tggatccttg
gtcgtttgtg ccttctctgg ttctcaccaa 1020gatgctatca agaagggatt ctccaaggct
gaagccagag ctgctagggg tgacaccaaa 1080tgggccattc catacttgcc attagaccct
aaggatatcg gtagaaacta cgaggccgtt 1140atcagagtca actctcaatc tggtaaggga
ggtgctgcct gggtcatctt gagatctctc 1200ggcttggact tgccaagaca cttgcaagtt
gtcttttctg gtattgttca ggaaagagct 1260gactctttgg gtagagaatt gaagtctgaa
gagattgccg ctttgttcaa cgagcagtac 1320tgctctactt ccaacttgtc tgtcaaggac
ttcgagataa ctaagagaaa gaatgctcca 1380gagaacaagg accgtgagat ctttgctgtc
ttgcaggctg gatccaagac cgttgacgtc 1440agtggacaag gtaacggacc tatttcggcc
tttgtggatg ccatatccaa gaaatacggt 1500gtttcctttg aagtcgtcaa ctacagtgaa
cacagtttag gcagtggtac ccagagtaag 1560gctgctactt acattgagtt agcctacaac
aactctaaca acgagcatgt tacaaagtgg 1620ggatgcggca ttaacacaga tgtgtcgcag
gcttcgatgg aggccattct ttctgttgtg 1680aactcattga ttgatagcaa ggaaattaat
ttgtag 171654571PRTPichia stipitis 54Met Leu
Lys Asp Pro Ser Thr Lys Tyr Ala Ala Phe Lys Gly Val Lys 1 5
10 15 Leu Asp Lys Arg Thr Trp Pro
Ser Lys Ser Ile Thr Lys Ala Pro Arg 20 25
30 Trp Leu Ser Thr Asp Leu Arg Asp Gly Asn Gln Ala
Leu Pro Asp Pro 35 40 45
Met Ser Val Glu Glu Lys Lys Glu Tyr Phe His Lys Leu Leu Glu Ile
50 55 60 Gly Phe Lys
Glu Ile Glu Val Ser Phe Pro Ser Ala Ser Gln Thr Asp 65
70 75 80 Phe Asp Phe Thr Arg Tyr Ala
Val Glu Asn Ala Pro Asp Asp Val Ser 85
90 95 Ile Gln Val Leu Thr Gln Ser Arg Glu Pro Leu
Ile Arg Arg Thr Val 100 105
110 Glu Ser Val Lys Gly Ala Lys Lys Ala Thr Ile His Thr Tyr Leu
Ala 115 120 125 Thr
Ser Asp Val Phe Arg Asp Val Val Phe Asn Met Ser Gln Glu Asp 130
135 140 Ala Ile Ala Lys Ala Ile
Glu Thr Thr Lys Leu Val Lys Ser Leu Thr 145 150
155 160 Lys Asp Asp Pro Glu Met Gln Glu Thr Glu Trp
Thr Leu Glu Phe Ser 165 170
175 Pro Glu Cys Phe Ser Asp Thr Pro Thr Glu Phe Ala Val Gln Ile Cys
180 185 190 Glu Ala
Val Lys Asn Val Trp Glu Pro Thr Val Glu Asn Pro Ile Ile 195
200 205 Phe Asn Leu Pro Ala Thr Val
Glu Val Ala Ser Pro Asn Val Tyr Ala 210 215
220 Asp Gln Ile Glu Tyr Phe Ala Thr His Ile Ser Glu
Arg Glu Lys Val 225 230 235
240 Cys Ile Ser Leu His Ala His Asn Asp Arg Gly Cys Gly Val Ala Ala
245 250 255 Ser Glu Leu
Gly Leu Leu Ala Gly Gly Asp Arg Val Glu Gly Cys Leu 260
265 270 Phe Gly Asn Gly Glu Arg Thr Gly
Asn Val Asp Leu Ile Thr Val Ala 275 280
285 Leu Asn Met Tyr Thr Asn Gly Val Ala Pro Glu Leu Asp
Phe Ser Glu 290 295 300
Ile Glu Lys Leu Ile Glu Val Ser Glu Arg Cys Asn Lys Ile Pro Val 305
310 315 320 His Pro Arg Ala
Pro Tyr Ser Gly Ser Leu Val Val Cys Ala Phe Ser 325
330 335 Gly Ser His Gln Asp Ala Ile Lys Lys
Gly Phe Ser Lys Ala Glu Ala 340 345
350 Arg Ala Ala Arg Gly Asp Thr Lys Trp Ala Ile Pro Tyr Leu
Pro Leu 355 360 365
Asp Pro Lys Asp Ile Gly Arg Asn Tyr Glu Ala Val Ile Arg Val Asn 370
375 380 Ser Gln Ser Gly Lys
Gly Gly Ala Ala Trp Val Ile Leu Arg Ser Leu 385 390
395 400 Gly Leu Asp Leu Pro Arg His Leu Gln Val
Val Phe Ser Gly Ile Val 405 410
415 Gln Glu Arg Ala Asp Ser Leu Gly Arg Glu Leu Lys Ser Glu Glu
Ile 420 425 430 Ala
Ala Leu Phe Asn Glu Gln Tyr Cys Ser Thr Ser Asn Leu Ser Val 435
440 445 Lys Asp Phe Glu Ile Thr
Lys Arg Lys Asn Ala Pro Glu Asn Lys Asp 450 455
460 Arg Glu Ile Phe Ala Val Leu Gln Ala Gly Ser
Lys Thr Val Asp Val 465 470 475
480 Ser Gly Gln Gly Asn Gly Pro Ile Ser Ala Phe Val Asp Ala Ile Ser
485 490 495 Lys Lys
Tyr Gly Val Ser Phe Glu Val Val Asn Tyr Ser Glu His Ser 500
505 510 Leu Gly Ser Gly Thr Gln Ser
Lys Ala Ala Thr Tyr Ile Glu Leu Ala 515 520
525 Tyr Asn Asn Ser Asn Asn Glu His Val Thr Lys Trp
Gly Cys Gly Ile 530 535 540
Asn Thr Asp Val Ser Gln Ala Ser Met Glu Ala Ile Leu Ser Val Val 545
550 555 560 Asn Ser Leu
Ile Asp Ser Lys Glu Ile Asn Leu 565 570
551728DNAPichia stipitis 55atgcctatgt tagctgatcc atcccaaaag tacaagcctt
tccctcctgt acatttaccc 60aatcgtcaat ggccatctcg tacgcttgaa aaaccaccta
gatggctttc caccgacttg 120agagatggaa accagtcctt accagatccc atgtcaattg
ctgaaaagaa ggagtacttc 180aagaagttgg tagatattgg tttcaaggaa atcgaagtgg
cctttccttc ggcatcgcag 240atcgactttg acttcaccag attcgctgtg gaaacagctc
cagcagacgt tgctgtccag 300gtcttgtctc cctgtagaga ggacttgatc aagcgtactg
tagagtcgtt gaccggagcc 360aaaaaggcta ttgtgcacat atacttggct acgtcggact
gcttccgtaa cgtagtcttt 420ggattaacca aggaagagtc gaaggctctt gccgtaaaat
gcgccaagtt ggtcagatct 480ttgactaaag atgaccccaa gcaacaagca actgagtggg
actttgagtt ctcgccagaa 540accttttcag acaccgatat ggactacgct gtagaggtct
gcgaagccgt caaggaggcc 600tggggcccta ctgaagacag acctatcatc ttcaacttgc
cagctaccgt ggaaatggct 660actccaaaca tatatgccga ccagatcgag tactttgcta
ctcacatctc ggaaagagaa 720aagatcgcaa tttcgttaca tccccacaac gacagaggat
gttctgttgc tgccgctgag 780ctcggccagt tagctggtgc tgacagagtt gagggatgct
tatttggaaa tggagaaaga 840acaggtaacg ttgacttggt caccttggct ctcaacttgt
atactcaggg tgtttcacct 900aagcttgact tctccgatat caactctgtt atcgacgtag
tagaaaagtg taacaagatt 960cctgttcatg caagagctcc ttatggaggt gcccttgtcg
tttgtgcctt cagtggatcg 1020caccaagacg ccatcaagaa aggtttcaat gtgcacgaga
agaaggtcga agctgctgca 1080ggaaaacatg tccactggca gttaccctac ttgccattgg
accctcagga tattggcaga 1140acttacgagg ccatcatcag agtcaactcg cagtctggta
agggtggttc cgcttgggtc 1200atcttgagaa acttggagct cgacttgccc agaggattgc
aagtggcttt ctccaaggtg 1260gttcaacagc gtgctgaagt caagggtcag gagttgacca
acgaagaatt gtgtgacttg 1320ttcaagcaag aatactacat tgactacgag ggtgacaact
tcaacgacca gacctacaag 1380ttgatcgact actccatcct gactcctgcc aagggccaga
aggaaattga agccgaaatc 1440cagatcgatg acaagatcgt caagatcaag ggccagggta
acggtcagct ttcggctttc 1500aatgctgccc tctccaaaca cctcaacatc gacttaaacg
tcaagcacta ccacgaacac 1560tcattgggtg tagactcgaa ttctcgtgca gccacctaca
tcgaagtctc actcaaaaac 1620gacaacgtca ccagatgggg tgtgggtatc catgaagatg
tctcgcaagc ttctttctta 1680tctctcatct ccatcttgaa cggcttgcac agaaacaagg
atatttag 172856575PRTPichia stipitis 56Met Pro Met Leu Ala
Asp Pro Ser Gln Lys Tyr Lys Pro Phe Pro Pro 1 5
10 15 Val His Leu Pro Asn Arg Gln Trp Pro Ser
Arg Thr Leu Glu Lys Pro 20 25
30 Pro Arg Trp Leu Ser Thr Asp Leu Arg Asp Gly Asn Gln Ser Leu
Pro 35 40 45 Asp
Pro Met Ser Ile Ala Glu Lys Lys Glu Tyr Phe Lys Lys Leu Val 50
55 60 Asp Ile Gly Phe Lys Glu
Ile Glu Val Ala Phe Pro Ser Ala Ser Gln 65 70
75 80 Ile Asp Phe Asp Phe Thr Arg Phe Ala Val Glu
Thr Ala Pro Ala Asp 85 90
95 Val Ala Val Gln Val Leu Ser Pro Cys Arg Glu Asp Leu Ile Lys Arg
100 105 110 Thr Val
Glu Ser Leu Thr Gly Ala Lys Lys Ala Ile Val His Ile Tyr 115
120 125 Leu Ala Thr Ser Asp Cys Phe
Arg Asn Val Val Phe Gly Leu Thr Lys 130 135
140 Glu Glu Ser Lys Ala Leu Ala Val Lys Cys Ala Lys
Leu Val Arg Ser 145 150 155
160 Leu Thr Lys Asp Asp Pro Lys Gln Gln Ala Thr Glu Trp Asp Phe Glu
165 170 175 Phe Ser Pro
Glu Thr Phe Ser Asp Thr Asp Met Asp Tyr Ala Val Glu 180
185 190 Val Cys Glu Ala Val Lys Glu Ala
Trp Gly Pro Thr Glu Asp Arg Pro 195 200
205 Ile Ile Phe Asn Leu Pro Ala Thr Val Glu Met Ala Thr
Pro Asn Ile 210 215 220
Tyr Ala Asp Gln Ile Glu Tyr Phe Ala Thr His Ile Ser Glu Arg Glu 225
230 235 240 Lys Ile Ala Ile
Ser Leu His Pro His Asn Asp Arg Gly Cys Ser Val 245
250 255 Ala Ala Ala Glu Leu Gly Gln Leu Ala
Gly Ala Asp Arg Val Glu Gly 260 265
270 Cys Leu Phe Gly Asn Gly Glu Arg Thr Gly Asn Val Asp Leu
Val Thr 275 280 285
Leu Ala Leu Asn Leu Tyr Thr Gln Gly Val Ser Pro Lys Leu Asp Phe 290
295 300 Ser Asp Ile Asn Ser
Val Ile Asp Val Val Glu Lys Cys Asn Lys Ile 305 310
315 320 Pro Val His Ala Arg Ala Pro Tyr Gly Gly
Ala Leu Val Val Cys Ala 325 330
335 Phe Ser Gly Ser His Gln Asp Ala Ile Lys Lys Gly Phe Asn Val
His 340 345 350 Glu
Lys Lys Val Glu Ala Ala Ala Gly Lys His Val His Trp Gln Leu 355
360 365 Pro Tyr Leu Pro Leu Asp
Pro Gln Asp Ile Gly Arg Thr Tyr Glu Ala 370 375
380 Ile Ile Arg Val Asn Ser Gln Ser Gly Lys Gly
Gly Ser Ala Trp Val 385 390 395
400 Ile Leu Arg Asn Leu Glu Leu Asp Leu Pro Arg Gly Leu Gln Val Ala
405 410 415 Phe Ser
Lys Val Val Gln Gln Arg Ala Glu Val Lys Gly Gln Glu Leu 420
425 430 Thr Asn Glu Glu Leu Cys Asp
Leu Phe Lys Gln Glu Tyr Tyr Ile Asp 435 440
445 Tyr Glu Gly Asp Asn Phe Asn Asp Gln Thr Tyr Lys
Leu Ile Asp Tyr 450 455 460
Ser Ile Ser Thr Pro Ala Lys Gly Gln Lys Glu Ile Glu Ala Glu Ile 465
470 475 480 Gln Ile Asp
Asp Lys Ile Val Lys Ile Lys Gly Gln Gly Asn Gly Gln 485
490 495 Leu Ser Ala Phe Asn Ala Ala Leu
Ser Lys His Leu Asn Ile Asp Leu 500 505
510 Asn Val Lys His Tyr His Glu His Ser Leu Gly Val Asp
Ser Asn Ser 515 520 525
Arg Ala Ala Thr Tyr Ile Glu Val Ser Leu Lys Asn Asp Asn Val Thr 530
535 540 Arg Trp Gly Val
Gly Ile His Glu Asp Val Ser Gln Ala Ser Phe Leu 545 550
555 560 Ser Leu Ile Ser Ile Leu Asn Gly Leu
His Arg Asn Lys Asp Ile 565 570
575 571101DNASaccharomyces cerevisiae 57atgttttcca gactgccaac
atcattggcc agaaatgttg cacgtcgtgc cccaacttct 60tttgtaagac cctctgcagc
agcagcagca ttgagattct catcaacaaa gacgatgacc 120gtcagagagg ccttgaatag
tgccatggcg gaagaattgg accgtgatga tgatgtcttc 180cttattggtg aagaagttgc
acaatataac ggggcttata aggtgtcaaa gggtttattg 240gacaggttcg gtgaacgtcg
tgtggttgac acacctatta ccgaatacgg gttcacaggt 300ttggccgttg gtgccgcttt
gaagggtttg aagccaattg tagagtttat gtcgttcaat 360ttctctatgc aagctatcga
tcatgttgtc aattccgctg caaagactca ctacatgtct 420ggtggtactc aaaaatgtca
aatggtcttc agaggtccta atggtgctgc agtgggtgtt 480ggtgctcaac attcacagga
cttttctcct tggtacggtt ccattccagg gttaaaggtc 540cttgtccctt attctgctga
agatgctagg ggtttgttaa aggccgccat cagagatcca 600aaccctgttg tatttttaga
gaacgaattg ttgtacggtg aatcttttga aatctcagaa 660gaagctttat cccctgagtt
caccttgcca tacaaggcta agatcgaaag agaaggtacc 720gatatttcca ttgttacgta
cacaagaaac gttcagtttt ctttggaagc cgctgaaatt 780ctacaaaaga aatatggtgt
ctctgcagaa gttatcaact tgcgttctat tagaccttta 840gatactgaag ctatcatcaa
aactgtcaag aagacaaacc acttgattac tgttgaatcc 900actttcccat catttggtgt
tggtgctgaa attgtcgccc aagttatgga gtctgaagcc 960tttgattact tggatgctcc
aatccaaaga gttactggtg ccgatgttcc aacaccttac 1020gctaaagaat tagaagattt
cgctttccct gatactccaa ccatcgttaa agctgtcaaa 1080gaagtcttgt caattgaata a
110158366PRTSaccharomyces
cerevisiae 58Met Phe Ser Arg Leu Pro Thr Ser Leu Ala Arg Asn Val Ala Arg
Arg 1 5 10 15 Ala
Pro Thr Ser Phe Val Arg Pro Ser Ala Ala Ala Ala Ala Leu Arg
20 25 30 Phe Ser Ser Thr Lys
Thr Met Thr Val Arg Glu Ala Leu Asn Ser Ala 35
40 45 Met Ala Glu Glu Leu Asp Arg Asp Asp
Asp Val Phe Leu Ile Gly Glu 50 55
60 Glu Val Ala Gln Tyr Asn Gly Ala Tyr Lys Val Ser Lys
Gly Leu Leu 65 70 75
80 Asp Arg Phe Gly Glu Arg Arg Val Val Asp Thr Pro Ile Thr Glu Tyr
85 90 95 Gly Phe Thr Gly
Leu Ala Val Gly Ala Ala Leu Lys Gly Leu Lys Pro 100
105 110 Ile Val Glu Phe Met Ser Phe Asn Phe
Ser Met Gln Ala Ile Asp His 115 120
125 Val Val Asn Ser Ala Ala Lys Thr His Tyr Met Ser Gly Gly
Thr Gln 130 135 140
Lys Cys Gln Met Val Phe Arg Gly Pro Asn Gly Ala Ala Val Gly Val 145
150 155 160 Gly Ala Gln His Ser
Gln Asp Phe Ser Pro Trp Tyr Gly Ser Ile Pro 165
170 175 Gly Leu Lys Val Leu Val Pro Tyr Ser Ala
Glu Asp Ala Arg Gly Leu 180 185
190 Leu Lys Ala Ala Ile Arg Asp Pro Asn Pro Val Val Phe Leu Glu
Asn 195 200 205 Glu
Leu Leu Tyr Gly Glu Ser Phe Glu Ile Ser Glu Glu Ala Leu Ser 210
215 220 Pro Glu Phe Thr Leu Pro
Tyr Lys Ala Lys Ile Glu Arg Glu Gly Thr 225 230
235 240 Asp Ile Ser Ile Val Thr Tyr Thr Arg Asn Val
Gln Phe Ser Leu Glu 245 250
255 Ala Ala Glu Ile Leu Gln Lys Lys Tyr Gly Val Ser Ala Glu Val Ile
260 265 270 Asn Leu
Arg Ser Ile Arg Pro Leu Asp Thr Glu Ala Ile Ile Lys Thr 275
280 285 Val Lys Lys Thr Asn His Leu
Ile Thr Val Glu Ser Thr Phe Pro Ser 290 295
300 Phe Gly Val Gly Ala Glu Ile Val Ala Gln Val Met
Glu Ser Glu Ala 305 310 315
320 Phe Asp Tyr Leu Asp Ala Pro Ile Gln Arg Val Thr Gly Ala Asp Val
325 330 335 Pro Thr Pro
Tyr Ala Lys Glu Leu Glu Asp Phe Ala Phe Pro Asp Thr 340
345 350 Pro Thr Ile Val Lys Ala Val Lys
Glu Val Leu Ser Ile Glu 355 360
365 59 1101DNASchizosaccharomyces pombe 59atgattcgtc ttcaaaagtt
tggtgaaatt gttgggacca gtcgttcttg gaaacttctt 60agttcaacca tcgcaaagcg
ctattcttct tcttccaatg gagtgaagga aatgaccgtt 120cgtgatgctt tgaacagtgc
aatggaagaa gaaatgaaac gtgacgatcg tgtcttcttg 180attggcgaag aggttgcgca
atacaatggt gcttataaga tatctagagg tttattagac 240aagtttggtc ctaaacgtgt
tatcgacact cccattactg aaatgggttt tactggtttg 300gcaacaggtg ctgcttttgc
tggtttacgt cctatttgtg agtttatgac tttcaatttt 360tccatgcagg ctatcgatca
tatcgttaac tcggccgcca gaaccctgta catgtctggt 420ggtattcagg cttgtcctat
tgtcttccgt ggacctaatg ggcctgccgc tgcagttgct 480gctcagcatt ctcaacactt
tgctccatgg tatggtagta tccctggtct taaagtagtt 540tctccttact cagcagaaga
tgctcgtggt ttgttgaagg ctgctattcg tgatcctaat 600cccgttgttg tacttgaaaa
cgaaattctt tatggtaaaa cctttccaat ttcgaaagaa 660gcgttgagcg aggactttgt
gcttcccttt ggccttgcta aggtggagcg ccccggtaaa 720gatatcacca tcgttggtga
gtctatttct gttgttactg ctttagaagc agctgacaag 780ctcaaggctg actatggtgt
tgaagctgaa gttataaact tgcgtagtat tcgtccttta 840gacatcaata ctatcgcggc
cagtgttaag aagacaaatc gtattgtgac tgttgaccag 900gcatatagtc aacatggtat
tggtagtgaa attgctgctc aaattatgga gtctgacgca 960tttgattatc ttgatgctcc
tgttgaacgt gtaagtatgg cagatgttcc catgccttat 1020agtcatcctg ttgaggctgc
ttctgtccca aatgccgatg ttgttgttgc tgctgctaaa 1080aaatgcttgt atattaaata a
110160366PRTSchizosaccharomyces pombe 60Met Ile Arg Leu Gln Lys Phe Gly
Glu Ile Val Gly Thr Ser Arg Ser 1 5 10
15 Trp Lys Leu Leu Ser Ser Thr Ile Ala Lys Arg Tyr Ser
Ser Ser Ser 20 25 30
Asn Gly Val Lys Glu Met Thr Val Arg Asp Ala Leu Asn Ser Ala Met
35 40 45 Glu Glu Glu Met
Lys Arg Asp Asp Arg Val Phe Leu Ile Gly Glu Glu 50
55 60 Val Ala Gln Tyr Asn Gly Ala Tyr
Lys Ile Ser Arg Gly Leu Leu Asp 65 70
75 80 Lys Phe Gly Pro Lys Arg Val Ile Asp Thr Pro Ile
Thr Glu Met Gly 85 90
95 Phe Thr Gly Leu Ala Thr Gly Ala Ala Phe Ala Gly Leu Arg Pro Ile
100 105 110 Cys Glu Phe
Met Thr Phe Asn Phe Ser Met Gln Ala Ile Asp His Ile 115
120 125 Val Asn Ser Ala Ala Arg Thr Leu
Tyr Met Ser Gly Gly Ile Gln Ala 130 135
140 Cys Pro Ile Val Phe Arg Gly Pro Asn Gly Pro Ala Ala
Ala Val Ala 145 150 155
160 Ala Gln His Ser Gln His Phe Ala Pro Trp Tyr Gly Ser Ile Pro Gly
165 170 175 Leu Lys Val Val
Ser Pro Tyr Ser Ala Glu Asp Ala Arg Gly Leu Leu 180
185 190 Lys Ala Ala Ile Arg Asp Pro Asn Pro
Val Val Val Leu Glu Asn Glu 195 200
205 Ile Leu Tyr Gly Lys Thr Phe Pro Ile Ser Lys Glu Ala Leu
Ser Glu 210 215 220
Asp Phe Val Leu Pro Phe Gly Leu Ala Lys Val Glu Arg Pro Gly Lys 225
230 235 240 Asp Ile Thr Ile Val
Gly Glu Ser Ile Ser Val Val Thr Ala Leu Glu 245
250 255 Ala Ala Asp Lys Leu Lys Ala Asp Tyr Gly
Val Glu Ala Glu Val Ile 260 265
270 Asn Leu Arg Ser Ile Arg Pro Leu Asp Ile Asn Thr Ile Ala Ala
Ser 275 280 285 Val
Lys Lys Thr Asn Arg Ile Val Thr Val Asp Gln Ala Tyr Ser Gln 290
295 300 His Gly Ile Gly Ser Glu
Ile Ala Ala Gln Ile Met Glu Ser Asp Ala 305 310
315 320 Phe Asp Tyr Leu Asp Ala Pro Val Glu Arg Val
Ser Met Ala Asp Val 325 330
335 Pro Met Pro Tyr Ser His Pro Val Glu Ala Ala Ser Val Pro Asn Ala
340 345 350 Asp Val
Val Val Ala Ala Ala Lys Lys Cys Leu Tyr Ile Lys 355
360 365 61 1140DNACandida albicans 61atgtcatcat
tatcatcagt caccaggagt gctaaattag ccactcaatc tttgaaatac 60aacactagac
catcattatc taaaattggt caatttcaaa catcaaaaat cacttatcgt 120gccaattcca
cacaatcaac tcctgtcaaa gaaattactg tcagagatgc tcttaaccaa 180gctttatctg
aagaattaga cagagatgaa gatgttttcc ttatgggtga agaagttgcc 240caatacaatg
gtgcctataa agtcagtaga ggattattgg acaaatttgg tgaaaagaga 300gttattgaca
ctccaattac tgaaatgggg ttcactggat tagctgttgg tgctgcttta 360catggtctta
aaccagtttt ggaatttatg acttggaatt ttgctatgca aggtattgat 420catattttaa
attctgctgc taaaactctt tatatgtctg gtggtaaaca accatgtaat 480ataactttcc
gtggtcctaa tggtgctgct gctggtgttg ctgctcaaca ttctcagtgt 540tatgctgctt
ggtatggttc aattcctggt ttaaaagttt tatctcctta ttctgctgaa 600gattataagg
gtttacttaa agctgccatt agagatccta acccagttgt tttcttggaa 660aatgaaattg
cttatggtga aacttttaaa gtttctgaag aattttcatc tccagatttc 720attttaccaa
ttggtaaagc caaaattgaa aaagaaggta ctgatttaac cattgttggt 780catagtcgtg
cccttaaatt tgccgttgaa gccgctgaaa ttttggaaaa agatttcgga 840attaaagctg
aagtgctcaa tttaagatca attaaaccat tggatgttcc agctattgtt 900gattcagtta
aaaagactaa tcatttggtt actgttgaaa atggattccc aggttttggt 960gttggttcag
aaatttgtgc tcaaattatg gaaagtgaag cctttgatta tttggatgct 1020ccagttgaaa
gagttactgg ttgtgaagtt ccaactccat atgctaaaga attggaagat 1080tttgctttcc
cagacactga agttatcttg agagcttgta aaaaagtatt aagtttgtaa
114062379PRTCandida albicans 62Met Ser Ser Leu Ser Ser Val Thr Arg Ser
Ala Lys Leu Ala Thr Gln 1 5 10
15 Ser Leu Lys Tyr Asn Thr Arg Pro Ser Leu Ser Lys Ile Gly Gln
Phe 20 25 30 Gln
Thr Ser Lys Ile Thr Tyr Arg Ala Asn Ser Thr Gln Ser Thr Pro 35
40 45 Val Lys Glu Ile Thr Val
Arg Asp Ala Leu Asn Gln Ala Leu Ser Glu 50 55
60 Glu Leu Asp Arg Asp Glu Asp Val Phe Leu Met
Gly Glu Glu Val Ala 65 70 75
80 Gln Tyr Asn Gly Ala Tyr Lys Val Ser Arg Gly Leu Leu Asp Lys Phe
85 90 95 Gly Glu
Lys Arg Val Ile Asp Thr Pro Ile Thr Glu Met Gly Phe Thr 100
105 110 Gly Leu Ala Val Gly Ala Ala
Leu His Gly Leu Lys Pro Val Leu Glu 115 120
125 Phe Met Thr Trp Asn Phe Ala Met Gln Gly Ile Asp
His Ile Leu Asn 130 135 140
Ser Ala Ala Lys Thr Leu Tyr Met Ser Gly Gly Lys Gln Pro Cys Asn 145
150 155 160 Ile Thr Phe
Arg Gly Pro Asn Gly Ala Ala Ala Gly Val Ala Ala Gln 165
170 175 His Ser Gln Cys Tyr Ala Ala Trp
Tyr Gly Ser Ile Pro Gly Leu Lys 180 185
190 Val Leu Ser Pro Tyr Ser Ala Glu Asp Tyr Lys Gly Leu
Leu Lys Ala 195 200 205
Ala Ile Arg Asp Pro Asn Pro Val Val Phe Leu Glu Asn Glu Ile Ala 210
215 220 Tyr Gly Glu Thr
Phe Lys Val Ser Glu Glu Phe Ser Ser Pro Asp Phe 225 230
235 240 Ile Leu Pro Ile Gly Lys Ala Lys Ile
Glu Lys Glu Gly Thr Asp Leu 245 250
255 Thr Ile Val Gly His Ser Arg Ala Leu Lys Phe Ala Val Glu
Ala Ala 260 265 270
Glu Ile Leu Glu Lys Asp Phe Gly Ile Lys Ala Glu Val Leu Asn Leu
275 280 285 Arg Ser Ile Lys
Pro Leu Asp Val Pro Ala Ile Val Asp Ser Val Lys 290
295 300 Lys Thr Asn His Leu Val Thr Val
Glu Asn Gly Phe Pro Gly Phe Gly 305 310
315 320 Val Gly Ser Glu Ile Cys Ala Gln Ile Met Glu Ser
Glu Ala Phe Asp 325 330
335 Tyr Leu Asp Ala Pro Val Glu Arg Val Thr Gly Cys Glu Val Pro Thr
340 345 350 Pro Tyr Ala
Lys Glu Leu Glu Asp Phe Ala Phe Pro Asp Thr Glu Val 355
360 365 Ile Leu Arg Ala Cys Lys Lys Val
Leu Ser Leu 370 375
631065DNAKluyveromyces lactis 63atgatgatgc tttctaacac ttttaagagg
gctgttcctt ctgtggttca atccatgaga 60tttgcttcta ccaagaccat gaccgtcaga
gaagctttga attctgccat ggccgaagaa 120atggaccgtg atgatgatgt tttcatcatt
ggtgaagaag ttgctcaata taatggtgct 180tacaaggtta ccaagggttt attggaccgt
ttcggtgaaa gaagagttgt tgacactcca 240attaccgaaa tgggtttcac tggtttggct
gttggtgccg ctttgaaggg tttaaagcca 300attgttgaat tcatgtcttt caacttctcc
atgcaagcta tggatcaagt cattaactcc 360gctgctaaga cttactatat gtccggtggt
actcagaaat gtcaaatcgt tttcagaggt 420ccaaacggtt ctgctgtcgg tgttgctgct
caacattccc aagattattc tgcttggtac 480ggttctgttc caggtatgaa ggttttggtt
ccatactctg ctgaagatgc tagaggtttg 540ttgaaggctg ccattcgtga tccaaaccca
gttgttttct tggaaaacga attgttatac 600ggtcaatctt tcgaagtctc tgaagaatct
ctgtctactg atttcacttt gccatacaaa 660gcaaaggttg aaagagaagg ttctgatatc
tctatcatca gttacaccag aaatgttcaa 720ttctctttgg aagctgctga aattttgtct
aagcaatacg gtgtttctgc tgaagttatc 780aatttgagag ccattagacc tttggatgtt
gaagctatca tcaacactgt caagaagacc 840aaccacttga ttactgttga atctactttc
ccagctttcg gtgttggtgc tgaaattatc 900gctcaaatta tggaatctga agccttcgat
tatttggatg ctccaattca aagagttact 960ggtgctgaag tcccaactcc ttatgctaag
gaattagaag attttgcttt cccagaccct 1020gacaccattg tcagagctgc taaaagtgtt
ttgtctattg aatga 106564354PRTKluyveromyces lactis 64Met
Met Met Leu Ser Asn Thr Phe Lys Arg Ala Val Pro Ser Val Val 1
5 10 15 Gln Ser Met Arg Phe Ala
Ser Thr Lys Thr Met Thr Val Arg Glu Ala 20
25 30 Leu Asn Ser Ala Met Ala Glu Glu Met Asp
Arg Asp Asp Asp Val Phe 35 40
45 Ile Ile Gly Glu Glu Val Ala Gln Tyr Asn Gly Ala Tyr Lys
Val Thr 50 55 60
Lys Gly Leu Leu Asp Arg Phe Gly Glu Arg Arg Val Val Asp Thr Pro 65
70 75 80 Ile Thr Glu Met Gly
Phe Thr Gly Leu Ala Val Gly Ala Ala Leu Lys 85
90 95 Gly Leu Lys Pro Ile Val Glu Phe Met Ser
Phe Asn Phe Ser Met Gln 100 105
110 Ala Met Asp Gln Val Ile Asn Ser Ala Ala Lys Thr Tyr Tyr Met
Ser 115 120 125 Gly
Gly Thr Gln Lys Cys Gln Ile Val Phe Arg Gly Pro Asn Gly Ser 130
135 140 Ala Val Gly Val Ala Ala
Gln His Ser Gln Asp Tyr Ser Ala Trp Tyr 145 150
155 160 Gly Ser Val Pro Gly Met Lys Val Leu Val Pro
Tyr Ser Ala Glu Asp 165 170
175 Ala Arg Gly Leu Leu Lys Ala Ala Ile Arg Asp Pro Asn Pro Val Val
180 185 190 Phe Leu
Glu Asn Glu Leu Leu Tyr Gly Gln Ser Phe Glu Val Ser Glu 195
200 205 Glu Ser Leu Ser Thr Asp Phe
Thr Leu Pro Tyr Lys Ala Lys Val Glu 210 215
220 Arg Glu Gly Ser Asp Ile Ser Ile Ile Ser Tyr Thr
Arg Asn Val Gln 225 230 235
240 Phe Ser Leu Glu Ala Ala Glu Ile Leu Ser Lys Gln Tyr Gly Val Ser
245 250 255 Ala Glu Val
Ile Asn Leu Arg Ala Ile Arg Pro Leu Asp Val Glu Ala 260
265 270 Ile Ile Asn Thr Val Lys Lys Thr
Asn His Leu Ile Thr Val Glu Ser 275 280
285 Thr Phe Pro Ala Phe Gly Val Gly Ala Glu Ile Ile Ala
Gln Ile Met 290 295 300
Glu Ser Glu Ala Phe Asp Tyr Leu Asp Ala Pro Ile Gln Arg Val Thr 305
310 315 320 Gly Ala Glu Val
Pro Thr Pro Tyr Ala Lys Glu Leu Glu Asp Phe Ala 325
330 335 Phe Pro Asp Pro Asp Thr Ile Val Arg
Ala Ala Lys Ser Val Leu Ser 340 345
350 Ile Glu 65993DNAYarrowia lipolytica 65atgactgtca
gagacgccct caacaccgca ctgcgagagg agatggaccg aaacgataat 60gttttcatca
tgggtgagga ggtcggccag tacaacggtg cctacaaggt caccaagggc 120cttctcgaca
agttcggcga gaagcgagtg gttgacaccc ctatcaccga gatgggtttc 180gccggtgttt
gtgtcggtgc cgccctggcc ggtctcaccc ccgtctgcga gttcatgacc 240tggaacttcg
ccatgcaggc cattgatcag atcatcaatt ccggtgccaa gacctactac 300atgtccggag
gtacccagca gtgcaatgtc accttccgag gtcctaacgg tgccgccgct 360ggtgttgctg
cccaacactc tcaggatttc accgggtggt acggccagat tcccggtctc 420aaggtcgtct
ctccctacag ctctgaggat gccaagggtc tgctcaaggc cgccatccga 480gaccccaacg
tgactgtttt cctcgagaac gagatcatgt acggagagtc tttccccatg 540tctgaggagg
ccatgtcccc cgacttcgtt ctgccccttg gaaaggccaa gattgagcga 600gagggtaagg
atatcactct tgtcggtcac tcccgaaacg tcgagaccgc cctcaaggcc 660gccgacctcc
tcaagaagca ccacaacgtc gatgccgagg tcattaacct gcgaactgtc 720aagcctctcg
acactgagac cattttcaac tccatcaaga agactaaccg acttgtctct 780gtcgaggctg
gcttccccgc ctttggcatg ggctccgagc tctgtggtgt cgtcaacgac 840tcctgggcct
gggattacct tgatgccccc atccagcgag ttaccggagc tgaggttccc 900actccttacg
ccattgagct tgagaacttc gccttcccca cacccgagat tgttgtcaag 960gctgccaagg
acgccctcta cattgaggag tag
99366330PRTYarrowia lipolytica 66Met Thr Val Arg Asp Ala Leu Asn Thr Ala
Leu Arg Glu Glu Met Asp 1 5 10
15 Arg Asn Asp Asn Val Phe Ile Met Gly Glu Glu Val Gly Gln Tyr
Asn 20 25 30 Gly
Ala Tyr Lys Val Thr Lys Gly Leu Leu Asp Lys Phe Gly Glu Lys 35
40 45 Arg Val Val Asp Thr Pro
Ile Thr Glu Met Gly Phe Ala Gly Val Cys 50 55
60 Val Gly Ala Ala Leu Ala Gly Leu Thr Pro Val
Cys Glu Phe Met Thr 65 70 75
80 Trp Asn Phe Ala Met Gln Ala Ile Asp Gln Ile Ile Asn Ser Gly Ala
85 90 95 Lys Thr
Tyr Tyr Met Ser Gly Gly Thr Gln Gln Cys Asn Val Thr Phe 100
105 110 Arg Gly Pro Asn Gly Ala Ala
Ala Gly Val Ala Ala Gln His Ser Gln 115 120
125 Asp Phe Thr Gly Trp Tyr Gly Gln Ile Pro Gly Leu
Lys Val Val Ser 130 135 140
Pro Tyr Ser Ser Glu Asp Ala Lys Gly Leu Leu Lys Ala Ala Ile Arg 145
150 155 160 Asp Pro Asn
Val Thr Val Phe Leu Glu Asn Glu Ile Met Tyr Gly Glu 165
170 175 Ser Phe Pro Met Ser Glu Glu Ala
Met Ser Pro Asp Phe Val Leu Pro 180 185
190 Leu Gly Lys Ala Lys Ile Glu Arg Glu Gly Lys Asp Ile
Thr Leu Val 195 200 205
Gly His Ser Arg Asn Val Glu Thr Ala Leu Lys Ala Ala Asp Leu Leu 210
215 220 Lys Lys His His
Asn Val Asp Ala Glu Val Ile Asn Leu Arg Thr Val 225 230
235 240 Lys Pro Leu Asp Thr Glu Thr Ile Phe
Asn Ser Ile Lys Lys Thr Asn 245 250
255 Arg Leu Val Ser Val Glu Ala Gly Phe Pro Ala Phe Gly Met
Gly Ser 260 265 270
Glu Leu Cys Gly Val Val Asn Asp Ser Trp Ala Trp Asp Tyr Leu Asp
275 280 285 Ala Pro Ile Gln
Arg Val Thr Gly Ala Glu Val Pro Thr Pro Tyr Ala 290
295 300 Ile Glu Leu Glu Asn Phe Ala Phe
Pro Thr Pro Glu Ile Val Val Lys 305 310
315 320 Ala Ala Lys Asp Ala Leu Tyr Ile Glu Glu
325 330 671170DNAPichia stipitis 67atggctccca
agttatccca gatcgcccag acggcccgct tggccgcttc ggccactaga 60gcccacaaca
tcgccaatgt gactggaaac actaccagat ccgtagccca agctggccag 120taccaggcat
tgagaatgat ggattcgcgt gccgcttcgt cgtcggctgt aggctcaaag 180accatcaccg
tcagagacgc ccttaatgcc gggcttgccg aggagttgga caaggacgac 240gatgtcttcc
tcatgggtga agaagtggcc caatacaacg gtgcctacaa ggtgtcacgt 300ggtttgttgg
atcgttttgg tgaaagacgt gtgattgata cccctatcac tgaaatgggt 360ttcactggtt
tggctgttgg agctgccctt catggtttga agcctgtgtt ggagttcatg 420accttcaact
tcgctatgca agctatcgat caaatcgtta actctgccgc taagacctat 480tacatgtccg
gaggtaaaca accgtgtaac atcaccttcc gtggtcccaa tggtgctgct 540gccggtgtcg
gtgctcaaca ttcgcaatgt tacgctgcat ggtatggatc tattcctggt 600ttgaaggttg
tttcgcccta ctctgccgag gactacaagg gtttgatcaa ggctgccatc 660agagacccta
acccagttgt gtttttggaa aacgaaatcg cctacggtga aaccttcgat 720atctccgagg
aagctctctc cacagacttt gttttgccta tcggcaaggc caatgtcgaa 780agagaaggaa
ctgacttgac atttgtatcg cattccagat ctgtcaagtt ctgtatggaa 840gccgctgaaa
ccttggagaa ggaatacggc gtcaaggccg aagtcatcaa cttgagatcc 900atcaagcctt
tggatgttcc taccattgtt gagtcagtca agaagactaa ccacttggtc 960actgttgaag
ccggattccc agcctttggt gttggttctg aaatctgtgc ccagatcatg 1020gaatccgagg
cttttgatta cttggatgct ccagtcgaaa gagtcactgg ttgcgaagtt 1080ccaactccat
atgctaagga attggaagac tttgctttcc cagacgaacc taccgtaatc 1140agagccgcca
aaaaggtgtt atctttgtaa
117068389PRTPichia stipitis 68Met Ala Pro Lys Leu Ser Gln Ile Ala Gln Thr
Ala Arg Leu Ala Ala 1 5 10
15 Ser Ala Thr Arg Ala His Asn Ile Ala Asn Val Thr Gly Asn Thr Thr
20 25 30 Arg Ser
Val Ala Gln Ala Gly Gln Tyr Gln Ala Leu Arg Met Met Asp 35
40 45 Ser Arg Ala Ala Ser Ser Ser
Ala Val Gly Ser Lys Thr Ile Thr Val 50 55
60 Arg Asp Ala Leu Asn Ala Gly Leu Ala Glu Glu Leu
Asp Lys Asp Asp 65 70 75
80 Asp Val Phe Leu Met Gly Glu Glu Val Ala Gln Tyr Asn Gly Ala Tyr
85 90 95 Lys Val Ser
Arg Gly Leu Leu Asp Arg Phe Gly Glu Arg Arg Val Ile 100
105 110 Asp Thr Pro Ile Thr Glu Met Gly
Phe Thr Gly Leu Ala Val Gly Ala 115 120
125 Ala Leu His Gly Leu Lys Pro Val Leu Glu Phe Met Thr
Phe Asn Phe 130 135 140
Ala Met Gln Ala Ile Asp Gln Ile Val Asn Ser Ala Ala Lys Thr Tyr 145
150 155 160 Tyr Met Ser Gly
Gly Lys Gln Pro Cys Asn Ile Thr Phe Arg Gly Pro 165
170 175 Asn Gly Ala Ala Ala Gly Val Gly Ala
Gln His Ser Gln Cys Tyr Ala 180 185
190 Ala Trp Tyr Gly Ser Ile Pro Gly Leu Lys Val Val Ser Pro
Tyr Ser 195 200 205
Ala Glu Asp Tyr Lys Gly Leu Ile Lys Ala Ala Ile Arg Asp Pro Asn 210
215 220 Pro Val Val Phe Leu
Glu Asn Glu Ile Ala Tyr Gly Glu Thr Phe Asp 225 230
235 240 Ile Ser Glu Glu Ala Leu Ser Thr Asp Phe
Val Leu Pro Ile Gly Lys 245 250
255 Ala Asn Val Glu Arg Glu Gly Thr Asp Leu Thr Phe Val Ser His
Ser 260 265 270 Arg
Ser Val Lys Phe Cys Met Glu Ala Ala Glu Thr Leu Glu Lys Glu 275
280 285 Tyr Gly Val Lys Ala Glu
Val Ile Asn Leu Arg Ser Ile Lys Pro Leu 290 295
300 Asp Val Pro Thr Ile Val Glu Ser Val Lys Lys
Thr Asn His Leu Val 305 310 315
320 Thr Val Glu Ala Gly Phe Pro Ala Phe Gly Val Gly Ser Glu Ile Cys
325 330 335 Ala Gln
Ile Met Glu Ser Glu Ala Phe Asp Tyr Leu Asp Ala Pro Val 340
345 350 Glu Arg Val Thr Gly Cys Glu
Val Pro Thr Pro Tyr Ala Lys Glu Leu 355 360
365 Glu Asp Phe Ala Phe Pro Asp Glu Pro Thr Val Ile
Arg Ala Ala Lys 370 375 380
Lys Val Leu Ser Leu 385 691263DNASaccharomyces
cerevisiae 69atgcttgctg cttcattcaa acgccaacca tcacaattgg tccgcgggtt
aggagctgtt 60cttcgcactc ccaccaggat aggtcatgtt cgtaccatgg caactttaaa
aacaactgat 120aagaaggccc ctgaggacat cgagggctcg gacacagtgc aaattgagtt
gcctgaatct 180tccttcgagt cgtatatgct agagcctcca gacttgtctt atgagacttc
gaaagccacc 240ttgttacaga tgtataaaga tatggtcatc atcagaagaa tggagatggc
ttgtgacgcc 300ttgtacaagg ccaagaaaat cagaggtttt tgccatctat ctgttggtca
ggaggccatt 360gctgtcggta tcgagaatgc catcacaaaa ttggattcca tcatcacatc
ttacagatgt 420cacggtttca cttttatgag aggtgcctca gtgaaagccg ttctggctga
attgatgggt 480agaagagccg gtgtctctta tggtaagggt ggttccatgc acctttacgc
tccaggcttc 540tatggtggta atggtatcgt gggtgcccag gttcctttag gtgcaggttt
agcttttgct 600caccaataca agaacgagga cgcctgctct ttcactttgt atggtgatgg
tgcctctaat 660caaggtcaag tttttgaatc tttcaacatg gccaaattat ggaatttgcc
cgtcgtgttt 720tgctgtgaga acaacaagta cggtatgggt accgccgctt caagatcctc
cgcgatgact 780gaatatttca agcgtggtca atatattcca ggtttaaaag ttaacggtat
ggatattcta 840gctgtctacc aagcatccaa gtttgctaag gactggtgtc tatccggcaa
aggtcctctc 900gttctagaat atgaaaccta taggtacggt ggccattcta tgtctgatcc
cggtactacc 960tacagaacta gagacgagat tcagcatatg agatccaaga acgatccaat
tgctggtctt 1020aagatgcatt tgattgatct aggtattgcc actgaagctg aagtcaaagc
ttacgacaag 1080tccgctagaa aatacgttga cgaacaagtt gaattagctg atgctgctcc
tcctccagaa 1140gccaaattat ccatcttgtt tgaagacgtc tacgtgaaag gtacagaaac
tccaacccta 1200agaggtagga tccctgaaga tacttgggac ttcaaaaagc aaggttttgc
ctctagggat 1260taa
126370420PRTSaccharomyces cerevisiae 70Met Leu Ala Ala Ser Phe
Lys Arg Gln Pro Ser Gln Leu Val Arg Gly 1 5
10 15 Leu Gly Ala Val Leu Arg Thr Pro Thr Arg Ile
Gly His Val Arg Thr 20 25
30 Met Ala Thr Leu Lys Thr Thr Asp Lys Lys Ala Pro Glu Asp Ile
Glu 35 40 45 Gly
Ser Asp Thr Val Gln Ile Glu Leu Pro Glu Ser Ser Phe Glu Ser 50
55 60 Tyr Met Leu Glu Pro Pro
Asp Leu Ser Tyr Glu Thr Ser Lys Ala Thr 65 70
75 80 Leu Leu Gln Met Tyr Lys Asp Met Val Ile Ile
Arg Arg Met Glu Met 85 90
95 Ala Cys Asp Ala Leu Tyr Lys Ala Lys Lys Ile Arg Gly Phe Cys His
100 105 110 Leu Ser
Val Gly Gln Glu Ala Ile Ala Val Gly Ile Glu Asn Ala Ile 115
120 125 Thr Lys Leu Asp Ser Ile Ile
Thr Ser Tyr Arg Cys His Gly Phe Thr 130 135
140 Phe Met Arg Gly Ala Ser Val Lys Ala Val Leu Ala
Glu Leu Met Gly 145 150 155
160 Arg Arg Ala Gly Val Ser Tyr Gly Lys Gly Gly Ser Met His Leu Tyr
165 170 175 Ala Pro Gly
Phe Tyr Gly Gly Asn Gly Ile Val Gly Ala Gln Val Pro 180
185 190 Leu Gly Ala Gly Leu Ala Phe Ala
His Gln Tyr Lys Asn Glu Asp Ala 195 200
205 Cys Ser Phe Thr Leu Tyr Gly Asp Gly Ala Ser Asn Gln
Gly Gln Val 210 215 220
Phe Glu Ser Phe Asn Met Ala Lys Leu Trp Asn Leu Pro Val Val Phe 225
230 235 240 Cys Cys Glu Asn
Asn Lys Tyr Gly Met Gly Thr Ala Ala Ser Arg Ser 245
250 255 Ser Ala Met Thr Glu Tyr Phe Lys Arg
Gly Gln Tyr Ile Pro Gly Leu 260 265
270 Lys Val Asn Gly Met Asp Ile Leu Ala Val Tyr Gln Ala Ser
Lys Phe 275 280 285
Ala Lys Asp Trp Cys Leu Ser Gly Lys Gly Pro Leu Val Leu Glu Tyr 290
295 300 Glu Thr Tyr Arg Tyr
Gly Gly His Ser Met Ser Asp Pro Gly Thr Thr 305 310
315 320 Tyr Arg Thr Arg Asp Glu Ile Gln His Met
Arg Ser Lys Asn Asp Pro 325 330
335 Ile Ala Gly Leu Lys Met His Leu Ile Asp Leu Gly Ile Ala Thr
Glu 340 345 350 Ala
Glu Val Lys Ala Tyr Asp Lys Ser Ala Arg Lys Tyr Val Asp Glu 355
360 365 Gln Val Glu Leu Ala Asp
Ala Ala Pro Pro Pro Glu Ala Lys Leu Ser 370 375
380 Ile Leu Phe Glu Asp Val Tyr Val Lys Gly Thr
Glu Thr Pro Thr Leu 385 390 395
400 Arg Gly Arg Ile Pro Glu Asp Thr Trp Asp Phe Lys Lys Gln Gly Phe
405 410 415 Ala Ser
Arg Asp 420 711230DNASchizosaccharomyces pombe 71atgtttcgaa
cttgtacgaa gattggaaca gttcccaagg ttcttgtgaa ccaaaagggc 60ttgatcgatg
gccttcgtcg ggtcaccaca gacgcaacca cttctcgtgc caatccggct 120catgtgcctg
aggaacatga caagccattt cctgttaaat tagatgatag tgtattcgaa 180ggatacaaga
tcgatgtccc ttctactgaa atcgaagtta caaagggaga gttattgggt 240ttgtacgaga
agatggtgac tattcgtcgt ctagaacttg catgcgatgc cttgtataag 300gctaagaaga
ttcgtggatt ctgtcatctt agcattggcc aagaagctgt agctgcagga 360attgaaggtg
ctattacact tgacgacagt attatcacat cttatagatg ccacggtttt 420gcttataccc
gtggtttgtc aattcgaagc attattggtg agctcatggg acgtcaatgt 480ggtgcttcca
agggcaaggg tggttctatg cacattttcg ccaaaaactt ctatggtggt 540aatggtattg
ttggtgctca aattcctttg ggtgctggta ttggtttcgc acagaagtat 600cttgaaaaac
ccactactac ttttgctcta tatggtgatg gtgcatctaa ccaaggtcaa 660gctttcgagg
ccttcaacat ggccaaatta tggggtcttc ccgttatttt tgcttgtgaa 720aacaacaaat
acggtatggg tactagtgct gaacgctctt ctgccatgac tgagttctac 780aaacgtggac
agtacattcc cggtcttttg gttaacggta tggatgtttt ggctgttttg 840caggcttcaa
agtttgctaa gaagtacact gttgaaaact ctcaacctct gcttatggaa 900tttgtgactt
atcgttatgg tggtcactcc atgtccgatc ccggtactac ttatcgtagc 960cgtgaagaag
tgcaaaaagt acgtgctgct agagatccta ttgagggttt gaagaagcac 1020atcatggagt
ggggcgtcgc taatgccaat gagcttaaaa acattgagaa gagaatccgt 1080ggtatggttg
atgaggaggt tcgtatcgct gaagaaagcc ctttccccga tcctattgag 1140gagagtttgt
tttcagatgt ttacgttgca ggaactgaac ccgcttacgc ccgtggtaga 1200aattccctgg
aatatcatca atataagtaa
123072409PRTSchizosaccharomyces pombe 72Met Phe Arg Thr Cys Thr Lys Ile
Gly Thr Val Pro Lys Val Leu Val 1 5 10
15 Asn Gln Lys Gly Leu Ile Asp Gly Leu Arg Arg Val Thr
Thr Asp Ala 20 25 30
Thr Thr Ser Arg Ala Asn Pro Ala His Val Pro Glu Glu His Asp Lys
35 40 45 Pro Phe Pro Val
Lys Leu Asp Asp Ser Val Phe Glu Gly Tyr Lys Ile 50
55 60 Asp Val Pro Ser Thr Glu Ile Glu
Val Thr Lys Gly Glu Leu Leu Gly 65 70
75 80 Leu Tyr Glu Lys Met Val Thr Ile Arg Arg Leu Glu
Leu Ala Cys Asp 85 90
95 Ala Leu Tyr Lys Ala Lys Lys Ile Arg Gly Phe Cys His Leu Ser Ile
100 105 110 Gly Gln Glu
Ala Val Ala Ala Gly Ile Glu Gly Ala Ile Thr Leu Asp 115
120 125 Asp Ser Ile Ile Thr Ser Tyr Arg
Cys His Gly Phe Ala Tyr Thr Arg 130 135
140 Gly Leu Ser Ile Arg Ser Ile Ile Gly Glu Leu Met Gly
Arg Gln Cys 145 150 155
160 Gly Ala Ser Lys Gly Lys Gly Gly Ser Met His Ile Phe Ala Lys Asn
165 170 175 Phe Tyr Gly Gly
Asn Gly Ile Val Gly Ala Gln Ile Pro Leu Gly Ala 180
185 190 Gly Ile Gly Phe Ala Gln Lys Tyr Leu
Glu Lys Pro Thr Thr Thr Phe 195 200
205 Ala Leu Tyr Gly Asp Gly Ala Ser Asn Gln Gly Gln Ala Phe
Glu Ala 210 215 220
Phe Asn Met Ala Lys Leu Trp Gly Leu Pro Val Ile Phe Ala Cys Glu 225
230 235 240 Asn Asn Lys Tyr Gly
Met Gly Thr Ser Ala Glu Arg Ser Ser Ala Met 245
250 255 Thr Glu Phe Tyr Lys Arg Gly Gln Tyr Ile
Pro Gly Leu Leu Val Asn 260 265
270 Gly Met Asp Val Leu Ala Val Leu Gln Ala Ser Lys Phe Ala Lys
Lys 275 280 285 Tyr
Thr Val Glu Asn Ser Gln Pro Leu Leu Met Glu Phe Val Thr Tyr 290
295 300 Arg Tyr Gly Gly His Ser
Met Ser Asp Pro Gly Thr Thr Tyr Arg Ser 305 310
315 320 Arg Glu Glu Val Gln Lys Val Arg Ala Ala Arg
Asp Pro Ile Glu Gly 325 330
335 Leu Lys Lys His Ile Met Glu Trp Gly Val Ala Asn Ala Asn Glu Leu
340 345 350 Lys Asn
Ile Glu Lys Arg Ile Arg Gly Met Val Asp Glu Glu Val Arg 355
360 365 Ile Ala Glu Glu Ser Pro Phe
Pro Asp Pro Ile Glu Glu Ser Leu Phe 370 375
380 Ser Asp Val Tyr Val Ala Gly Thr Glu Pro Ala Tyr
Ala Arg Gly Arg 385 390 395
400 Asn Ser Leu Glu Tyr His Gln Tyr Lys 405
731206DNACandida albicans 73atgtaccgtg caacagctac tagtcgccaa
ttggtcggta ctaccgccaa tatacttgtc 60gccaaaagat caatggccaa agccgcctca
gatttggtca ctatcgaatt accagccagc 120tcctacgaag gatacaattt ggaagttcca
gctttgagtt ttgaaaccga aaaagaaacc 180ttattgaaaa tgtacaaaga tatgattatc
atcagaagaa tggaaatggc agccgatgct 240ttatacaaga gtaaaaaaat tagaggtttc
tgtcacttgt ctgtcggtca agaagccatt 300gctgttggta ttgaaaatgc cattacacca
actgacactg tcattacctc ttatagatgt 360cacggttttg cattcatgag aggtgcttct
gtcaaatctg ttttggccga gttaatgggt 420agaagatctg gtattgccaa cggtaagggt
ggatcaatgc atatgttcac taacggattc 480tacggtggta acggtattgt tggtgcccaa
gttccattgg gtgctggatt ggctttctcc 540cacaagtaca agaacgacaa agctgtcact
tttgatttgt atggtgatgg tgcgtctaac 600caaggacaag ttttcgaagc ttacaacatg
gccaaattgt ggaacttacc agttattttc 660gcctgtgaaa acaacaagta tggtatgggt
acctctgctg ccagatcatc agctatgacc 720gaatactaca agagaggtca atatatccca
ggtttgaaaa tcaacggtat ggatgtgttg 780gccacctacc aagcctccaa attcgccaaa
gactgggctt ctcaaggcaa tggacctctt 840gttttagaat acgaaactta cagatatggt
ggtcactcca tgtctgatcc aggtaccact 900tacagaacca gagaagaagt ccaacatatg
agatctagaa acgatccaat tgctggattg 960aaagctgttt tgttagaaaa agagattgct
tctgaagacg aaatcaaatc ttacgacaaa 1020gccgctagaa aatacgttga tgaacaagtt
gctgctgctg aagctgatgc tccaccagaa 1080gctaaaatgg atattttatt cgaagacgtt
tatgttccag gtagtgagat tcctgttttg 1140agaggtagaa tctccgacga tagttgggat
ttcaaaaaca agactttttt gaacaaggtc 1200tattaa
120674401PRTCandida albicans 74Met Tyr
Arg Ala Thr Ala Thr Ser Arg Gln Leu Val Gly Thr Thr Ala 1 5
10 15 Asn Ile Leu Val Ala Lys Arg
Ser Met Ala Lys Ala Ala Ser Asp Leu 20 25
30 Val Thr Ile Glu Leu Pro Ala Ser Ser Tyr Glu Gly
Tyr Asn Leu Glu 35 40 45
Val Pro Ala Leu Ser Phe Glu Thr Glu Lys Glu Thr Leu Leu Lys Met
50 55 60 Tyr Lys Asp
Met Ile Ile Ile Arg Arg Met Glu Met Ala Ala Asp Ala 65
70 75 80 Leu Tyr Lys Ser Lys Lys Ile
Arg Gly Phe Cys His Leu Ser Val Gly 85
90 95 Gln Glu Ala Ile Ala Val Gly Ile Glu Asn Ala
Ile Thr Pro Thr Asp 100 105
110 Thr Val Ile Thr Ser Tyr Arg Cys His Gly Phe Ala Phe Met Arg
Gly 115 120 125 Ala
Ser Val Lys Ser Val Leu Ala Glu Leu Met Gly Arg Arg Ser Gly 130
135 140 Ile Ala Asn Gly Lys Gly
Gly Ser Met His Met Phe Thr Asn Gly Phe 145 150
155 160 Tyr Gly Gly Asn Gly Ile Val Gly Ala Gln Val
Pro Leu Gly Ala Gly 165 170
175 Leu Ala Phe Ser His Lys Tyr Lys Asn Asp Lys Ala Val Thr Phe Asp
180 185 190 Leu Tyr
Gly Asp Gly Ala Ser Asn Gln Gly Gln Val Phe Glu Ala Tyr 195
200 205 Asn Met Ala Lys Leu Trp Asn
Leu Pro Val Ile Phe Ala Cys Glu Asn 210 215
220 Asn Lys Tyr Gly Met Gly Thr Ser Ala Ala Arg Ser
Ser Ala Met Thr 225 230 235
240 Glu Tyr Tyr Lys Arg Gly Gln Tyr Ile Pro Gly Leu Lys Ile Asn Gly
245 250 255 Met Asp Val
Leu Ala Thr Tyr Gln Ala Ser Lys Phe Ala Lys Asp Trp 260
265 270 Ala Ser Gln Gly Asn Gly Pro Leu
Val Leu Glu Tyr Glu Thr Tyr Arg 275 280
285 Tyr Gly Gly His Ser Met Ser Asp Pro Gly Thr Thr Tyr
Arg Thr Arg 290 295 300
Glu Glu Val Gln His Met Arg Ser Arg Asn Asp Pro Ile Ala Gly Leu 305
310 315 320 Lys Ala Val Leu
Leu Glu Lys Glu Ile Ala Ser Glu Asp Glu Ile Lys 325
330 335 Ser Tyr Asp Lys Ala Ala Arg Lys Tyr
Val Asp Glu Gln Val Ala Ala 340 345
350 Ala Glu Ala Asp Ala Pro Pro Glu Ala Lys Met Asp Ile Leu
Phe Glu 355 360 365
Asp Val Tyr Val Pro Gly Ser Glu Ile Pro Val Leu Arg Gly Arg Ile 370
375 380 Ser Asp Asp Ser Trp
Asp Phe Lys Asn Lys Thr Phe Leu Asn Lys Val 385 390
395 400 Tyr 751227DNAKluyveromyces lactis
75atgctatctt tgaaagctca atcctctgtg gttgggaagt ccagctcttt gagattggtt
60agaaactttt ctaaaaacgt ccgtgctttg tcccaggttg ctgatgaaac taagccaggt
120gatgatgacc tagttcaaat tgatttgcca gaaacctctt ttgaaggtta tcttttggat
180gttcctgaat taagttatca aaccaccaag tccaatttgc tacaaatgta caaggatatg
240attatcgtta gaagaatgga aatggcctgt gacgctttgt acaaggctaa gaaaattaga
300ggtttctgtc actcctctgt cggtcaagaa gccattgccg ttggtattga aaacgctatc
360actaagcgtg ataccgtcat cacctcttac agatgtcatg gtttcaccta catgagaggt
420gctgctgttc aagctgtgtt ggctgaattg atgggtagaa gaactggtgt gtccttcggt
480aagggtggtt ccatgcactt gtacgcccct ggtttctacg gtggtaatgg tatcgttggt
540gcccaagtcc cattgggtgc tggtttggcc ttcgctcatc aatacaaaca cgaagatgct
600tgttcttttg ccttgtacgg tgatggtgcc tctaaccaag gtcaagtttt cgaatccttc
660aacatggcca agttatggaa cttaccagcc gtcttctgtt gtgaaaacaa caagtacggt
720atgggtaccg ctgccgcaag atcttcagcc atgactgaat acttcaagcg tggtcaatac
780attcctggtt tgaaggttaa cggtatggat atcttggctg ttaccaagct taaggactgg
840actgtctccg gtaacggtcc aatcgttctt gaatacgaaa cttacagata tggtggtcac
900tctatgtctg atccaggtac tacttacaga accagagatg aaatccaaca catgagatct
960aagaacgatc caattgcagg tttaaagatg cacttattgg aattgggtat cgccacggaa
1020gatgaaatta aggcttacga caaggctgct agaaagtacg tcgatgagca agtcgaatta
1080gctgatgctg ccccagctcc agaagctaag atgtccatct tgttcgagga tgtctacgtt
1140ccaggttctg aaactccaac cctaagaggt agattgcaag aagatacttg ggattttgct
1200aagaagagct ttgctttcag agattag
122776408PRTKluyveromyces lactis 76Met Leu Ser Leu Lys Ala Gln Ser Ser
Val Val Gly Lys Ser Ser Ser 1 5 10
15 Leu Arg Leu Val Arg Asn Phe Ser Lys Asn Val Arg Ala Leu
Ser Gln 20 25 30
Val Ala Asp Glu Thr Lys Pro Gly Asp Asp Asp Leu Val Gln Ile Asp
35 40 45 Leu Pro Glu Thr
Ser Phe Glu Gly Tyr Leu Leu Asp Val Pro Glu Leu 50
55 60 Ser Tyr Gln Thr Thr Lys Ser Asn
Leu Leu Gln Met Tyr Lys Asp Met 65 70
75 80 Ile Ile Val Arg Arg Met Glu Met Ala Cys Asp Ala
Leu Tyr Lys Ala 85 90
95 Lys Lys Ile Arg Gly Phe Cys His Ser Ser Val Gly Gln Glu Ala Ile
100 105 110 Ala Val Gly
Ile Glu Asn Ala Ile Thr Lys Arg Asp Thr Val Ile Thr 115
120 125 Ser Tyr Arg Cys His Gly Phe Thr
Tyr Met Arg Gly Ala Ala Val Gln 130 135
140 Ala Val Leu Ala Glu Leu Met Gly Arg Arg Thr Gly Val
Ser Phe Gly 145 150 155
160 Lys Gly Gly Ser Met His Leu Tyr Ala Pro Gly Phe Tyr Gly Gly Asn
165 170 175 Gly Ile Val Gly
Ala Gln Val Pro Leu Gly Ala Gly Leu Ala Phe Ala 180
185 190 His Gln Tyr Lys His Glu Asp Ala Cys
Ser Phe Ala Leu Tyr Gly Asp 195 200
205 Gly Ala Ser Asn Gln Gly Gln Val Phe Glu Ser Phe Asn Met
Ala Lys 210 215 220
Leu Trp Asn Leu Pro Ala Val Phe Cys Cys Glu Asn Asn Lys Tyr Gly 225
230 235 240 Met Gly Thr Ala Ala
Ala Arg Ser Ser Ala Met Thr Glu Tyr Phe Lys 245
250 255 Arg Gly Gln Tyr Ile Pro Gly Leu Lys Val
Asn Gly Met Asp Ile Leu 260 265
270 Ala Val Thr Lys Leu Lys Asp Trp Thr Val Ser Gly Asn Gly Pro
Ile 275 280 285 Val
Leu Glu Tyr Glu Thr Tyr Arg Tyr Gly Gly His Ser Met Ser Asp 290
295 300 Pro Gly Thr Thr Tyr Arg
Thr Arg Asp Glu Ile Gln His Met Arg Ser 305 310
315 320 Lys Asn Asp Pro Ile Ala Gly Leu Lys Met His
Leu Leu Glu Leu Gly 325 330
335 Ile Ala Thr Glu Asp Glu Ile Lys Ala Tyr Asp Lys Ala Ala Arg Lys
340 345 350 Tyr Val
Asp Glu Gln Val Glu Leu Ala Asp Ala Ala Pro Ala Pro Glu 355
360 365 Ala Lys Met Ser Ile Leu Phe
Glu Asp Val Tyr Val Pro Gly Ser Glu 370 375
380 Thr Pro Thr Leu Arg Gly Arg Leu Gln Glu Asp Thr
Trp Asp Phe Ala 385 390 395
400 Lys Lys Ser Phe Ala Phe Arg Asp 405
771185DNAYarrowia lipolytica 77atgctcactg ccgctcgacg atctacacgg
ctcaccagcc gactcggcca ccaggtccga 60gcatactcca tcgctgacga tgccgacaag
aaatgcacaa tcacgctcaa ggaggattct 120tacaccacct acatgcttga ttctccccct
cctctcgagt tcgagatgac caagggtgag 180cttctgcaaa tgtacaagga catggtgacc
gtccgacgac tcgagatggc tgctgatgcc 240ctctacaagg ccaagaagat ccgaggtttc
tgccatctgt ctactggtca ggaggctgtt 300gccgtcggta tcgagaaggc catcgaccac
gacgattctg tcatcaccgc ctaccgatgc 360cacggtttcg cctacatgcg aggtgcctct
gtccgagcaa tcatcgccga gctgctcgga 420aagcgaaccg gtgtctccta cggtaagggt
ggatccatgc acatgttcac cgagggtttc 480tacggaggaa acggtattgt cggagcccag
gtccccgtcg gagctggtct cgccttcgcc 540cacaagtacc tcgagcagac cggaaaggcc
acctttgccc tgtacggtga cggtgcttcc 600aaccagggtc agatcttcga ggcctacaac
atggccaagc tctgggacct cccctgcatc 660tttgcatgcg agaacaacaa gtacggaatg
ggtaccgctg ctgctcgatc ctctgccctg 720acgcagtact acaagcgagg tcagtacatt
cccggtctca aggttaacgg aatggacatt 780ctgtccgtct accagggagc caagttcgcc
aaggagtgga ccacacacgg caagggtccc 840ctcgtcatgg agttcgagac ctaccgatac
ggtggtcact ccatgtccga tcccggaacc 900acctaccgaa cccgagagga gatccagtac
atgcgatccc acaacgatcc tatttctggt 960ctcaaggccc acatcctgga gcttaatttc
gccactgagg acgagcttaa gtctgtggac 1020aaggctgctc gagctatggt tgacaaggag
gttgcccttg ctgagtccga ccctgctcct 1080gaggctactg ccaaggttct gtttgaggat
atctacgttc ccggcaccga gcctcctgtg 1140atccgaggcc gaatcccttc cgaggactac
tactttaaga actaa 118578394PRTYarrowia lipolytica 78Met
Leu Thr Ala Ala Arg Arg Ser Thr Arg Leu Thr Ser Arg Leu Gly 1
5 10 15 His Gln Val Arg Ala Tyr
Ser Ile Ala Asp Asp Ala Asp Lys Lys Cys 20
25 30 Thr Ile Thr Leu Lys Glu Asp Ser Tyr Thr
Thr Tyr Met Leu Asp Ser 35 40
45 Pro Pro Pro Leu Glu Phe Glu Met Thr Lys Gly Glu Leu Leu
Gln Met 50 55 60
Tyr Lys Asp Met Val Thr Val Arg Arg Leu Glu Met Ala Ala Asp Ala 65
70 75 80 Leu Tyr Lys Ala Lys
Lys Ile Arg Gly Phe Cys His Leu Ser Thr Gly 85
90 95 Gln Glu Ala Val Ala Val Gly Ile Glu Lys
Ala Ile Asp His Asp Asp 100 105
110 Ser Val Ile Thr Ala Tyr Arg Cys His Gly Phe Ala Tyr Met Arg
Gly 115 120 125 Ala
Ser Val Arg Ala Ile Ile Ala Glu Leu Leu Gly Lys Arg Thr Gly 130
135 140 Val Ser Tyr Gly Lys Gly
Gly Ser Met His Met Phe Thr Glu Gly Phe 145 150
155 160 Tyr Gly Gly Asn Gly Ile Val Gly Ala Gln Val
Pro Val Gly Ala Gly 165 170
175 Leu Ala Phe Ala His Lys Tyr Leu Glu Gln Thr Gly Lys Ala Thr Phe
180 185 190 Ala Leu
Tyr Gly Asp Gly Ala Ser Asn Gln Gly Gln Ile Phe Glu Ala 195
200 205 Tyr Asn Met Ala Lys Leu Trp
Asp Leu Pro Cys Ile Phe Ala Cys Glu 210 215
220 Asn Asn Lys Tyr Gly Met Gly Thr Ala Ala Ala Arg
Ser Ser Ala Leu 225 230 235
240 Thr Gln Tyr Tyr Lys Arg Gly Gln Tyr Ile Pro Gly Leu Lys Val Asn
245 250 255 Gly Met Asp
Ile Leu Ser Val Tyr Gln Gly Ala Lys Phe Ala Lys Glu 260
265 270 Trp Thr Thr His Gly Lys Gly Pro
Leu Val Met Glu Phe Glu Thr Tyr 275 280
285 Arg Tyr Gly Gly His Ser Met Ser Asp Pro Gly Thr Thr
Tyr Arg Thr 290 295 300
Arg Glu Glu Ile Gln Tyr Met Arg Ser His Asn Asp Pro Ile Ser Gly 305
310 315 320 Leu Lys Ala His
Ile Leu Glu Leu Asn Phe Ala Thr Glu Asp Glu Leu 325
330 335 Lys Ser Val Asp Lys Ala Ala Arg Ala
Met Val Asp Lys Glu Val Ala 340 345
350 Leu Ala Glu Ser Asp Pro Ala Pro Glu Ala Thr Ala Lys Val
Leu Phe 355 360 365
Glu Asp Ile Tyr Val Pro Gly Thr Glu Pro Pro Val Ile Arg Gly Arg 370
375 380 Ile Pro Ser Glu Asp
Tyr Tyr Phe Lys Asn 385 390
791191DNAPichia stipitis 79atgttacgta ctgctgctgt tcgtcctctt aagggcggtg
ttgtcatcgc cagaagagcc 60atggcctcgt ccagcgactt ggtcagcatc gaattgcctg
aatcgtcgtt tgaaggctac 120aacttggaga tccccgagtt gactttcgaa accgaaaagg
aaaccttgtt gaagatgtac 180aaggatatga tcatcatcag aagaatggaa atggcttcag
acgccttgta caaggccaag 240aagatcagag ggttctgcca cttgtctgtt ggtcaagaag
ccattgccgt tggaattgag 300aacgccatta ctcctgaaga tactgtcatc acctcttaca
gatgtcacgg ttttgctttc 360atgagaggtg cttctgtcaa ggaagttctc ggagaattga
tgggtaagag atctggtgtt 420tcttatggta aaggtggttc tatgcacatg tttgccccag
gcttttacgg aggaaacggt 480atcgttggag ctcaagttcc attgggtgct ggtttagctt
tctcccacaa gtacagggga 540cagaaggctg ctgccttcac tttgtacggt gacggtgcct
ccaaccaggg acaagttttc 600gaagcctaca acatggccaa gttgtggaac ttgccttgta
tctttgcctg tgaaaacaac 660aagtacggta tgggtactgc tgctgccaga tcctctgcta
ttactgagta ctacaagaga 720ggtcaataca ttcctggttt gaagatcaac ggtatggacg
ttttggctac ctaccaggct 780tccaagtttg ccaaggactg ggctgctcaa ggcaacggac
cattggtttt ggaatacgaa 840acctacagat acggtggtca ctccatgtct gacccaggta
ccacctacag aacaagagaa 900gaagtgcaac acatgagatc cagaaacgat cctattgccg
gcttaaaggc tactttgttg 960gacaagggca ttgctaccga agaagaaatc aagtcctatg
acaaggctgc cagaaagtac 1020gtcgacgaac aagtcgctgc tgctgaagct gacgctcctc
ctgaagccaa gatggacatc 1080ttattcgaag atgtatatgt cccaggatct gaaatcccag
tgttgagagg cagaatctcg 1140gacgactcgt gggacttcaa gaacaaaact ttcttgaaca
aggtctacta g 119180396PRTPichia stipitis 80Met Leu Arg Thr Ala
Ala Val Arg Pro Leu Lys Gly Gly Val Val Ile 1 5
10 15 Ala Arg Arg Ala Met Ala Ser Ser Ser Asp
Leu Val Ser Ile Glu Leu 20 25
30 Pro Glu Ser Ser Phe Glu Gly Tyr Asn Leu Glu Ile Pro Glu Leu
Thr 35 40 45 Phe
Glu Thr Glu Lys Glu Thr Leu Leu Lys Met Tyr Lys Asp Met Ile 50
55 60 Ile Ile Arg Arg Met Glu
Met Ala Ser Asp Ala Leu Tyr Lys Ala Lys 65 70
75 80 Lys Ile Arg Gly Phe Cys His Leu Ser Val Gly
Gln Glu Ala Ile Ala 85 90
95 Val Gly Ile Glu Asn Ala Ile Thr Pro Glu Asp Thr Val Ile Thr Ser
100 105 110 Tyr Arg
Cys His Gly Phe Ala Phe Met Arg Gly Ala Ser Val Lys Glu 115
120 125 Val Leu Gly Glu Leu Met Gly
Lys Arg Ser Gly Val Ser Tyr Gly Lys 130 135
140 Gly Gly Ser Met His Met Phe Ala Pro Gly Phe Tyr
Gly Gly Asn Gly 145 150 155
160 Ile Val Gly Ala Gln Val Pro Leu Gly Ala Gly Leu Ala Phe Ser His
165 170 175 Lys Tyr Arg
Gly Gln Lys Ala Ala Ala Phe Thr Leu Tyr Gly Asp Gly 180
185 190 Ala Ser Asn Gln Gly Gln Val Phe
Glu Ala Tyr Asn Met Ala Lys Leu 195 200
205 Trp Asn Leu Pro Cys Ile Phe Ala Cys Glu Asn Asn Lys
Tyr Gly Met 210 215 220
Gly Thr Ala Ala Ala Arg Ser Ser Ala Ile Thr Glu Tyr Tyr Lys Arg 225
230 235 240 Gly Gln Tyr Ile
Pro Gly Leu Lys Ile Asn Gly Met Asp Val Leu Ala 245
250 255 Thr Tyr Gln Ala Ser Lys Phe Ala Lys
Asp Trp Ala Ala Gln Gly Asn 260 265
270 Gly Pro Leu Val Leu Glu Tyr Glu Thr Tyr Arg Tyr Gly Gly
His Ser 275 280 285
Met Ser Asp Pro Gly Thr Thr Tyr Arg Thr Arg Glu Glu Val Gln His 290
295 300 Met Arg Ser Arg Asn
Asp Pro Ile Ala Gly Leu Lys Ala Thr Leu Leu 305 310
315 320 Asp Lys Gly Ile Ala Thr Glu Glu Glu Ile
Lys Ser Tyr Asp Lys Ala 325 330
335 Ala Arg Lys Tyr Val Asp Glu Gln Val Ala Ala Ala Glu Ala Asp
Ala 340 345 350 Pro
Pro Glu Ala Lys Met Asp Ile Leu Phe Glu Asp Val Tyr Val Pro 355
360 365 Gly Ser Glu Ile Pro Val
Leu Arg Gly Arg Ile Ser Asp Asp Ser Trp 370 375
380 Asp Phe Lys Asn Lys Thr Phe Leu Asn Lys Val
Tyr 385 390 395 8176DNAartificial
sequenceprimer 81gtgagtgcta caagccacat ttaaactaag tcaattacac aaagttagtg
ggtcgcctga 60cgcatatacc tttttc
768276DNAartificial sequenceprimer 82attttaccta acaagttgtt
gcgtaaattt ataaagtaaa ttgtcggttt ttttgtgtgg 60tgccctcctc cttgtc
768333DNAartificial
sequenceprimer 83gagtcatctc aaacatatgt ctgcagatac ttc
338431DNAartificial sequenceprimer 84gaaatagctt taagaacctt
aatggcttcg g 31854280DNAartificial
sequencevector 85ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg cgtaatcatg
gtcatagctg 60tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
cggaagcata 120aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
gttgcgctca 180ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
cggccaacgc 240gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
tgactcgctg 300cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
aatacggtta 360tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
gcaaaaggcc 420aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
ccctgacgag 480catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
ataaagatac 540caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
gccgcttacc 600ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
ctcacgctgt 660aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
cgaacccccc 720gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
cccggtaaga 780cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
gaggtatgta 840ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
aaggacagta 900tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
tagctcttga 960tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
gcagattacg 1020cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
tgacgctcag 1080tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
gatcttcacc 1140tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
tgagtaaact 1200tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
ctgtctattt 1260cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
ggagggctta 1320ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
tccagattta 1380tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
aactttatcc 1440gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
gccagttaat 1500agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
gtcgtttggt 1560atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
ccccatgttg 1620tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
gttggccgca 1680gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
gccatccgta 1740agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
gtgtatgcgg 1800cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
tagcagaact 1860ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
gatcttaccg 1920ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
agcatctttt 1980actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
aaaaaaggga 2040ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
ttattgaagc 2100atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
gaaaaataaa 2160caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
agaaaccatt 2220attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
tctcgcgcgt 2280ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
cacagcttgt 2340ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
tgttggcggg 2400tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
gcaccatatg 2460cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
ccattcgcca 2520ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
attacgccag 2580ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
gttttcccag 2640tcacgacgtt gtaaaacgac ggccagtgaa ttcgagctcg gtacccccgg
ctctgagaca 2700gtagtaggtt agtcatcgct ctaccgacgc gcaggaaaag aaagaagcat
tgcggattac 2760gtattctaat gttcagcccg cggaacgcca gcaaatcacc acccatgcgc
atgatactga 2820gtcttgtaca cgctgggctt ccagtgtact gagagtgcac cataccacag
cttttcaatt 2880caattcatca tttttttttt attctttttt ttgatttcgg tttctttgaa
atttttttga 2940ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
gattggtata 3000tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
cccaactgca 3060cagaacaaaa acctgcagga aacgaagata aatcatgtcg aaagctacat
ataaggaacg 3120tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
acgaaaagca 3180aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
agttagttga 3240agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
ctgatttttc 3300catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
ttttactctt 3360cgaagacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
ctgcgggtgt 3420atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
gcccaggtat 3480tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
gccttttgat 3540gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
agggtactgt 3600tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
gagacatggg 3660tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
tagatgacaa 3720gggagacgca ttgggtcaac agtatagaac cgtggatgat gtggtctcta
caggatctga 3780cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
tagagggtga 3840acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
aaaactaaaa 3900aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
caatttaatt 3960atatcagtta ttaccctatg cggtgtgaaa taccgcacag atgcgtaagg
agaaaatacc 4020gcatcaggaa attgtaaacg ttaatatttt gttaaaattc gcgttaaatt
tttgttaaat 4080cagctcattt tttaaccaat aggccgaaat cggcaaaatc ttcagcccgc
ggaacgccag 4140caaatcacca cccatgcgca tgatactgag tcttgtacac gctgggcttc
cagtgatgat 4200acaacgagtt agccaaggtg agcacggatg tctaaattag aattacgttt
taatatcttt 4260ttttccatat ctagggctag
42808685DNAartificial sequenceprimer 86ccggctctga gacagtagta
ggttagtcat cgctctaccg acgcgcagga aaagaaagaa 60gcattgcgga ttacgtattc
taatg 858786DNAartificial
sequenceprimer 87ctagccctag atatggaaaa aaagatatta aaacgtaatt ctaatttaga
catccgtgct 60caccttggct aactcgttgt atcatc
868822DNAartificial sequenceprimer 88gagaagatgc ggccagcaaa
ac 228930DNAartificial
sequenceprimer 89cgccaaacaa gtttcgggtc accccacacg
309029DNAartificial sequenceprimer 90ctcaaaattc tattgtgttt
gccggtacc 299185DNAartificial
sequenceprimer 91cattagaata cgtaatccgc aatgcttctt tcttttcctg cgcgtcggta
gagcgatcgg 60tgaatgtctg gccgaacact aattc
859287DNAartificial sequenceprimer 92gatgatacaa cgagttagcc
aaggtgagca cggatgtcta aattagaatt acgtttgtta 60ccgctccatt agatggtacc
attttag 879330DNAartificial
sequenceprimer 93atagatcgtg gaaaactttc actacaaagc
309461DNAartificial sequenceprimer 94gaattagtgt tcggccagac
attcaccgat cgctctaccg acgcgcagga aaagaaagaa 60g
619560DNAartificial
sequenceprimer 95ctaaaatggt accatctaat ggagcggtaa caaacgtaat tctaatttag
acatccgtgc 609620DNAartificial sequenceprimer 96cccttggggc cgctaattag
209730DNAartificial
sequenceprimer 97gcattgcgga ttacgtattc taatgttcag
309864DNAartificial sequenceprimer 98cgtcggaggg ctgtcgcccg
ctcggcggct tctaatcacc ttggctaact cgttgtatca 60tcac
64998459DNAartificial
sequencevector 99ggtggagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
gcgtaatcat 60ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
aacataggag 120ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gaggtaactc
acattaattg 180cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
cattaatgaa 240tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
tcctcgctca 300ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
tcaaaggcgg 360taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
gcaaaaggcc 420agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
aggctccgcc 480cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
ccgacaggac 540tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
gttccgaccc 600tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
ctttctcata 660gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
ggctgtgtgc 720acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
cttgagtcca 780acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
attagcagag 840cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
ggctacacta 900gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
aaaagagttg 960gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
gtttgcaagc 1020agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
tctacggggt 1080ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
ttatcaaaaa 1140ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
taaagtatat 1200atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
atctcagcga 1260tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
actacgatac 1320gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
cgctcaccgg 1380ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
agtggtcctg 1440caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
gtaagtagtt 1500cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
gtgtcacgct 1560cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
gttacatgat 1620cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
gtcagaagta 1680agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
cttactgtca 1740tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
ttctgagaat 1800agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
accgcgccac 1860atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
aaactctcaa 1920ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
aactgatctt 1980cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
caaaatgccg 2040caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
ctttttcaat 2100attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
gaatgtattt 2160agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
cctgaacgaa 2220gcatctgtgc ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat
ttttcaaaca 2280aagaatctga gctgcatttt tacagaacag aaatgcaacg cgaaagcgct
attttaccaa 2340cgaagaatct gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg
ctaatttttc 2400aaacaaagaa tctgagctgc atttttacag aacagaaatg caacgcgaga
gcgctatttt 2460accaacaaag aatctatact tcttttttgt tctacaaaaa tgcatcccga
gagcgctatt 2520tttctaacaa agcatcttag attacttttt ttctcctttg tgcgctctat
aatgcagtct 2580cttgataact ttttgcactg taggtccgtt aaggttagaa gaaggctact
ttggtgtcta 2640ttttctcttc cataaaaaaa gcctgactcc acttcccgcg tttactgatt
actagcgaag 2700ctgcgggtgc attttttcaa gataaaggca tccccgatta tattctatac
cgatgtggat 2760tgcgcatact ttgtgaacag aaagtgatag cgttgatgat tcttcattgg
tcagaaaatt 2820atgaacggtt tcttctattt tgtctctata tactacgtat aggaaatgtt
tacattttcg 2880tattgttttc gattcactct atgaatagtt cttactacaa tttttttgtc
taaagagtaa 2940tactagagat aaacataaaa aatgtagagg tcgagtttag atgcaagttc
aaggagcgaa 3000aggtggatgg gtaggttata tagggatata gcacagagat atatagcaaa
gagatacttt 3060tgagcaatgt ttgtggaagc ggtattcgca atattttagt agctcgttac
agtccggtgc 3120gtttttggtt ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa
gcgctctgaa 3180gttcctatac tttctagaga ataggaactt cggaatagga acttcaaagc
gtttccgaaa 3240acgagcgctt ccgaaaatgc aacgcgagct gcgcacatac agctcactgt
tcacgtcgca 3300cctatatctg cgtgttgcct gtatatatat atacatgaga agaacggcat
agtgcgtgtt 3360tatgcttaaa tgcgtactta tatgcgtcta tttatgtagg atgaaaggta
gtctagtacc 3420tcctgtgata ttatcccatt ccatgcgggg tatcgtatgc ttccttcagc
actacccttt 3480agctgttcta tatgctgcca ctcctcaatt ggattagtct catccttcaa
tgctatcatt 3540tcctttgata ttggatcata ctaagaaacc attattatca tgacattaac
ctataaaaat 3600aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
aaacctctga 3660cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
gagcagacaa 3720gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
ctatgcggca 3780tcagagcaga ttgtactgag agtgcaccat accacagctt ttcaattcaa
ttcatcattt 3840tttttttatt cttttttttg atttcggttt ctttgaaatt tttttgattc
ggtaatctcc 3900gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat
acgcatatgt 3960agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag
aacaaaaacc 4020tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc
tgctactcat 4080cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac
aaacttgtgt 4140gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc
attaggtccc 4200aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat
ggagggcaca 4260gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga
agacagaaaa 4320tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata
cagaatagca 4380gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt
tagcggtttg 4440aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt
agcagaattg 4500tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga
cattgcgaag 4560agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg
aagagatgaa 4620ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg
agacgcattg 4680ggtcaacagt atagaaccgt ggatgatgtg gtctctacag gatctgacat
tattattgtt 4740ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg
ttacagaaaa 4800gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac
tgtattataa 4860gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata
tcagttatta 4920ccctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
tcaggaaatt 4980gtaaacgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
ctcatttttt 5040aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
cgagataggg 5100ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
ctccaacgtc 5160aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
accctaatca 5220agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
gagcccccga 5280tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
gaaagcgaaa 5340ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
caccacaccc 5400gccgcgctta atgcgccgct acagggcgcg tcgcgccatt cgccattcag
gctgcgcaac 5460tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc
gaaaggggga 5520tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg
acgttgtaaa 5580acgacggcca gtgagcgcgc gtaatacgac tcactatagg gcgaattggg
taccgggccc 5640cccctcgagg tcgacggtat cgataagctt gatatcgaat tcctgcagcc
cgggggatcc 5700gcatgcttgc atttagtcgt gcaatgtatg actttaagat ttgtgagcag
gaagaaaagg 5760gagaatcttc taacgataaa cccttgaaaa actgggtaga ctacgctatg
ttgagttgct 5820acgcaggctg cacaattaca cgagaatgct cccgcctagg atttaaggct
aagggacgtg 5880caatgcagac gacagatcta aatgaccgtg tcggtgaagt gttcgccaaa
cttttcggtt 5940aacacatgca gtgatgcacg cgcgatggtg ctaagttaca tatatatata
tatatatata 6000tatagccata gtgatgtcta agtaaccttt atggtatatt tcttaatgtg
gaaagatact 6060agcgcgcgca cccacacaca agcttcgtct tttcttgaag aaaagaggaa
gctcgctaaa 6120tgggattcca ctttccgttc cctgccagct gatggaaaaa ggttagtgga
acgatgaaga 6180ataaaaagag agatccactg aggtgaaatt tcagctgaca gcgagtttca
tgatcgtgat 6240gaacaatggt aacgagttgt ggctgttgcc agggagggtg gttctcaact
tttaatgtat 6300ggccaaatcg ctacttgggt ttgttatata acaaagaaga aataatgaac
tgattctctt 6360cctccttctt gtcctttctt aattctgttg taattacctt cctttgtaat
tttttttgta 6420attattcttc ttaataatcc aaacaaacac acatattaca atagctgagg
atgtatactg 6480tgggggatta cctgctggat cgcctgcacg aactggggat tgaagaaatt
ttcggtgtgc 6540caggcgatta taacctgcag ttcctggacc agattatctc gcacaaagat
atgaagtggg 6600tcggtaacgc caacgaactg aacgcgagct atatggcaga tggttatgcc
cgtaccaaaa 6660aagctgctgc gtttctgacg acctttggcg ttggcgaact gagcgccgtc
aacggactgg 6720caggaagcta cgccgagaac ctgccagttg tcgaaattgt tgggtcgcct
acttctaagg 6780ttcagaatga aggcaaattt gtgcaccata ctctggctga tggggatttt
aaacatttta 6840tgaaaatgca tgaaccggtt actgcggccc gcacgctgct gacagcagag
aatgctacgg 6900ttgagatcga ccgcgtcctg tctgcgctgc tgaaagagcg caagccggta
tatatcaatc 6960tgcctgtcga tgttgccgca gcgaaagccg aaaagccgtc gctgccactg
aaaaaagaaa 7020acagcacctc caatacatcg gaccaggaaa ttctgaataa aatccaggaa
tcactgaaga 7080atgcgaagaa accgatcgtc atcaccggac atgagatcat ctcttttggc
ctggaaaaaa 7140cggtcacgca gttcatttct aagaccaaac tgcctatcac caccctgaac
ttcggcaaat 7200ctagcgtcga tgaagcgctg ccgagttttc tgggtatcta taatggtacc
ctgtccgaac 7260cgaacctgaa agaattcgtc gaaagcgcgg actttatcct gatgctgggc
gtgaaactga 7320cggatagctc cacaggcgca tttacccacc atctgaacga gaataaaatg
atttccctga 7380atatcgacga aggcaaaatc tttaacgagc gcatccagaa cttcgatttt
gaatctctga 7440ttagttcgct gctggatctg tccgaaattg agtataaagg taaatatatt
gataaaaaac 7500aggaggattt tgtgccgtct aatgcgctgc tgagtcagga tcgtctgtgg
caagccgtag 7560aaaacctgac acagtctaat gaaacgattg ttgcggaaca gggaacttca
tttttcggcg 7620cctcatccat ttttctgaaa tccaaaagcc atttcattgg ccaaccgctg
tgggggagta 7680ttggttatac ctttccggcg gcgctgggtt cacagattgc agataaggaa
tcacgccatc 7740tgctgtttat tggtgacggc agcctgcagc tgactgtcca ggaactgggg
ctggcgatcc 7800gtgaaaaaat caatccgatt tgctttatca tcaataacga cggctacacc
gtcgaacgcg 7860aaattcatgg accgaatcaa agttacaatg acatcccgat gtggaactat
agcaaactgc 7920cggaatcctt tggcgcgaca gaggatcgcg tggtgagtaa aattgtgcgt
acggaaaacg 7980aatttgtgtc ggttatgaaa gaagcgcagg ctgacccgaa tcgcatgtat
tggattgaac 8040tgatcctggc aaaagaaggc gcaccgaaag ttctgaaaaa gatggggaaa
ctgtttgcgg 8100agcaaaataa aagctaatta attaagagta agcgaatttc ttatgattta
tgatttttat 8160tattaaataa gttataaaaa aaataagtgt atacaaattt taaagtgact
cttaggtttt 8220aaaacgaaaa ttcttattct tgagtaactc tttcctgtag gtcaggttgc
tttctcaggt 8280atagcatgag gtcgctctta ttgaccacac ctctaccggc atgccgagca
aatgcctgca 8340aatcgctccc catttcaccc aattgtagat atgctaactc cagcaatgag
ttgatgaatc 8400tcggtgtgta ttttatgtcc tcagaggaca acacctgtgg tactagttct
agagcggcc 845910064DNAartificial sequenceprimer 100gtgatgatac
aacgagttag ccaaggtgat tagaagccgc cgagcgggcg acagccctcc 60gacg
6410137DNAartificial sequenceprimer 101ttttctcctt gacgttaaag tatagaggta
tattaac 3710288DNAartificial sequenceprimer
102tgtacacacg taatcgcgcg tgtacatgtc tatatgtgtt acttgaacta tactgttttg
60gcattgcgga ttacgtattc taatgttc
8810397DNAartificial sequenceprimer 103aacagctcct aacccgcgga ccaattgtga
tggttggcgt ttgaatgaag cagcaagcat 60ttttctcctt gacgttaaag tatagaggta
tattaac 9710430DNAartificial sequenceprimer
104gattcaggca actcaatttg cactgtgtcc
301051449DNASaccharomyces cerevisiaeCDS(1)..(1449) 105atg tct gcc ttt gtc
agg gtg gtt cca aga ata tcc aga agt tca gta 48Met Ser Ala Phe Val
Arg Val Val Pro Arg Ile Ser Arg Ser Ser Val 1 5
10 15 ctc acc aga tca ttg
aga ctg caa ttg aga tgc tac gca tcg tac cca 96Leu Thr Arg Ser Leu
Arg Leu Gln Leu Arg Cys Tyr Ala Ser Tyr Pro 20
25 30 gag cac acc att att
ggt atg ccg gca ctg tct cct acg atg acg caa 144Glu His Thr Ile Ile
Gly Met Pro Ala Leu Ser Pro Thr Met Thr Gln 35
40 45 ggt aat ctt gct gct
tgg act aag aag gaa ggt gac caa ttg tct ccc 192Gly Asn Leu Ala Ala
Trp Thr Lys Lys Glu Gly Asp Gln Leu Ser Pro 50
55 60 ggt gaa gtt att gcc
gaa ata gaa aca gac aag gct caa atg gac ttt 240Gly Glu Val Ile Ala
Glu Ile Glu Thr Asp Lys Ala Gln Met Asp Phe 65
70 75 80 gag ttc caa gaa gat
ggt tac tta gcc aag att cta gtt cct gaa ggt 288Glu Phe Gln Glu Asp
Gly Tyr Leu Ala Lys Ile Leu Val Pro Glu Gly 85
90 95 aca aag gac att cct
gtc aac aag cct att gcc gtc tat gtg gag gac 336Thr Lys Asp Ile Pro
Val Asn Lys Pro Ile Ala Val Tyr Val Glu Asp 100
105 110 aaa gct gat gtg cca
gct ttt aag gac ttt aag ctg gag gat tca ggt 384Lys Ala Asp Val Pro
Ala Phe Lys Asp Phe Lys Leu Glu Asp Ser Gly 115
120 125 tct gat tca aag acc
agt acg aag gct cag cct gcc gaa cca cag gca 432Ser Asp Ser Lys Thr
Ser Thr Lys Ala Gln Pro Ala Glu Pro Gln Ala 130
135 140 gaa aag aaa caa gaa
gcg cca gct gaa gag acc aag act tct gca cct 480Glu Lys Lys Gln Glu
Ala Pro Ala Glu Glu Thr Lys Thr Ser Ala Pro 145
150 155 160 gaa gct aag aaa tct
gac gtt gct gct cct caa ggt agg att ttt gcc 528Glu Ala Lys Lys Ser
Asp Val Ala Ala Pro Gln Gly Arg Ile Phe Ala 165
170 175 tct cca ctt gcc aag
act atc gcc ttg gaa aag ggt att tct ttg aag 576Ser Pro Leu Ala Lys
Thr Ile Ala Leu Glu Lys Gly Ile Ser Leu Lys 180
185 190 gat gtt cac ggc act
gga ccc cgc ggt aga att acc aag gct gac att 624Asp Val His Gly Thr
Gly Pro Arg Gly Arg Ile Thr Lys Ala Asp Ile 195
200 205 gag tca tat cta gaa
aag tcg tct aag cag tct tct caa acc agt ggt 672Glu Ser Tyr Leu Glu
Lys Ser Ser Lys Gln Ser Ser Gln Thr Ser Gly 210
215 220 gct gcc gcc gcc act
cct gcc gcc gct acc tca agc act act gct ggc 720Ala Ala Ala Ala Thr
Pro Ala Ala Ala Thr Ser Ser Thr Thr Ala Gly 225
230 235 240 tct gct cca tcg cct
tct tct aca gca tca tat gag gat gtt cca att 768Ser Ala Pro Ser Pro
Ser Ser Thr Ala Ser Tyr Glu Asp Val Pro Ile 245
250 255 tca acc atg aga agc
atc att gga gaa cgt tta ttg caa tct act caa 816Ser Thr Met Arg Ser
Ile Ile Gly Glu Arg Leu Leu Gln Ser Thr Gln 260
265 270 ggc att cca tca tac
atc gtt tcc tcc aag ata tcc atc tcc aaa ctt 864Gly Ile Pro Ser Tyr
Ile Val Ser Ser Lys Ile Ser Ile Ser Lys Leu 275
280 285 ttg aaa ttg aga cag
tcc ttg aac gct aca gca aac gac aag tac aaa 912Leu Lys Leu Arg Gln
Ser Leu Asn Ala Thr Ala Asn Asp Lys Tyr Lys 290
295 300 ctg tcc att aat gac
cta tta gta aaa gcc atc act gtt gcg gct aag 960Leu Ser Ile Asn Asp
Leu Leu Val Lys Ala Ile Thr Val Ala Ala Lys 305
310 315 320 agg gtg cca gat gcc
aat gcc tac tgg tta cct aat gag aac gtt atc 1008Arg Val Pro Asp Ala
Asn Ala Tyr Trp Leu Pro Asn Glu Asn Val Ile 325
330 335 cgt aaa ttc aag aat
gtc gat gtc tca gtc gct gtt gcc aca cca aca 1056Arg Lys Phe Lys Asn
Val Asp Val Ser Val Ala Val Ala Thr Pro Thr 340
345 350 gga tta ttg aca cca
att gtc aag aat tgt gag gcc aag ggc ttg tcg 1104Gly Leu Leu Thr Pro
Ile Val Lys Asn Cys Glu Ala Lys Gly Leu Ser 355
360 365 caa atc tct aac gaa
atc aag gaa cta gtc aag cgt gcc aga ata aac 1152Gln Ile Ser Asn Glu
Ile Lys Glu Leu Val Lys Arg Ala Arg Ile Asn 370
375 380 aaa ttg gca cca gag
gaa ttc caa ggt ggg acc att tgc ata tcc aat 1200Lys Leu Ala Pro Glu
Glu Phe Gln Gly Gly Thr Ile Cys Ile Ser Asn 385
390 395 400 atg ggc atg aat aat
gct gtt aac atg ttt act tcg att atc aac cca 1248Met Gly Met Asn Asn
Ala Val Asn Met Phe Thr Ser Ile Ile Asn Pro 405
410 415 cca cag tct aca atc
ttg gcc atc gct act gtt gaa agg gtc gct gtg 1296Pro Gln Ser Thr Ile
Leu Ala Ile Ala Thr Val Glu Arg Val Ala Val 420
425 430 gaa gac gcc gct gct
gag aac gga ttc tcc ttt gat aac cag gtt acc 1344Glu Asp Ala Ala Ala
Glu Asn Gly Phe Ser Phe Asp Asn Gln Val Thr 435
440 445 ata aca ggg acc ttt
gat cat aga acc att gat ggc gcc aaa ggt gca 1392Ile Thr Gly Thr Phe
Asp His Arg Thr Ile Asp Gly Ala Lys Gly Ala 450
455 460 gaa ttc atg aag gaa
ttg aaa act gtt att gaa aat cct ttg gaa atg 1440Glu Phe Met Lys Glu
Leu Lys Thr Val Ile Glu Asn Pro Leu Glu Met 465
470 475 480 cta ttg tga
1449Leu Leu
106482PRTSaccharomyces cerevisiae 106Met Ser Ala Phe Val Arg Val Val Pro
Arg Ile Ser Arg Ser Ser Val 1 5 10
15 Leu Thr Arg Ser Leu Arg Leu Gln Leu Arg Cys Tyr Ala Ser
Tyr Pro 20 25 30
Glu His Thr Ile Ile Gly Met Pro Ala Leu Ser Pro Thr Met Thr Gln
35 40 45 Gly Asn Leu Ala
Ala Trp Thr Lys Lys Glu Gly Asp Gln Leu Ser Pro 50
55 60 Gly Glu Val Ile Ala Glu Ile Glu
Thr Asp Lys Ala Gln Met Asp Phe 65 70
75 80 Glu Phe Gln Glu Asp Gly Tyr Leu Ala Lys Ile Leu
Val Pro Glu Gly 85 90
95 Thr Lys Asp Ile Pro Val Asn Lys Pro Ile Ala Val Tyr Val Glu Asp
100 105 110 Lys Ala Asp
Val Pro Ala Phe Lys Asp Phe Lys Leu Glu Asp Ser Gly 115
120 125 Ser Asp Ser Lys Thr Ser Thr Lys
Ala Gln Pro Ala Glu Pro Gln Ala 130 135
140 Glu Lys Lys Gln Glu Ala Pro Ala Glu Glu Thr Lys Thr
Ser Ala Pro 145 150 155
160 Glu Ala Lys Lys Ser Asp Val Ala Ala Pro Gln Gly Arg Ile Phe Ala
165 170 175 Ser Pro Leu Ala
Lys Thr Ile Ala Leu Glu Lys Gly Ile Ser Leu Lys 180
185 190 Asp Val His Gly Thr Gly Pro Arg Gly
Arg Ile Thr Lys Ala Asp Ile 195 200
205 Glu Ser Tyr Leu Glu Lys Ser Ser Lys Gln Ser Ser Gln Thr
Ser Gly 210 215 220
Ala Ala Ala Ala Thr Pro Ala Ala Ala Thr Ser Ser Thr Thr Ala Gly 225
230 235 240 Ser Ala Pro Ser Pro
Ser Ser Thr Ala Ser Tyr Glu Asp Val Pro Ile 245
250 255 Ser Thr Met Arg Ser Ile Ile Gly Glu Arg
Leu Leu Gln Ser Thr Gln 260 265
270 Gly Ile Pro Ser Tyr Ile Val Ser Ser Lys Ile Ser Ile Ser Lys
Leu 275 280 285 Leu
Lys Leu Arg Gln Ser Leu Asn Ala Thr Ala Asn Asp Lys Tyr Lys 290
295 300 Leu Ser Ile Asn Asp Leu
Leu Val Lys Ala Ile Thr Val Ala Ala Lys 305 310
315 320 Arg Val Pro Asp Ala Asn Ala Tyr Trp Leu Pro
Asn Glu Asn Val Ile 325 330
335 Arg Lys Phe Lys Asn Val Asp Val Ser Val Ala Val Ala Thr Pro Thr
340 345 350 Gly Leu
Leu Thr Pro Ile Val Lys Asn Cys Glu Ala Lys Gly Leu Ser 355
360 365 Gln Ile Ser Asn Glu Ile Lys
Glu Leu Val Lys Arg Ala Arg Ile Asn 370 375
380 Lys Leu Ala Pro Glu Glu Phe Gln Gly Gly Thr Ile
Cys Ile Ser Asn 385 390 395
400 Met Gly Met Asn Asn Ala Val Asn Met Phe Thr Ser Ile Ile Asn Pro
405 410 415 Pro Gln Ser
Thr Ile Leu Ala Ile Ala Thr Val Glu Arg Val Ala Val 420
425 430 Glu Asp Ala Ala Ala Glu Asn Gly
Phe Ser Phe Asp Asn Gln Val Thr 435 440
445 Ile Thr Gly Thr Phe Asp His Arg Thr Ile Asp Gly Ala
Lys Gly Ala 450 455 460
Glu Phe Met Lys Glu Leu Lys Thr Val Ile Glu Asn Pro Leu Glu Met 465
470 475 480 Leu Leu
1071500DNASaccharomyces cerevisiaeCDS(1)..(1500) 107atg tta aga atc aga
tca ctc cta aat aat aag cgt gcc ttt tcg tcc 48Met Leu Arg Ile Arg
Ser Leu Leu Asn Asn Lys Arg Ala Phe Ser Ser 1 5
10 15 aca gtc agg aca ttg
acc att aac aag tca cat gat gta gtc atc atc 96Thr Val Arg Thr Leu
Thr Ile Asn Lys Ser His Asp Val Val Ile Ile 20
25 30 ggt ggt ggc cct gct
ggt tac gtg gct gct atc aaa gct gct caa ttg 144Gly Gly Gly Pro Ala
Gly Tyr Val Ala Ala Ile Lys Ala Ala Gln Leu 35
40 45 gga ttt aac act gca
tgt gta gaa aaa aga ggc aaa tta ggc ggt acc 192Gly Phe Asn Thr Ala
Cys Val Glu Lys Arg Gly Lys Leu Gly Gly Thr 50
55 60 tgt ctt aac gtt gga
tgt atc ccc tcc aaa gca ctt cta aat aat tct 240Cys Leu Asn Val Gly
Cys Ile Pro Ser Lys Ala Leu Leu Asn Asn Ser 65
70 75 80 cat tta ttc cac caa
atg cat acg gaa gcg caa aag aga ggt att gac 288His Leu Phe His Gln
Met His Thr Glu Ala Gln Lys Arg Gly Ile Asp 85
90 95 gtc aac ggt gat atc
aaa att aac gta gca aac ttc caa aag gct aag 336Val Asn Gly Asp Ile
Lys Ile Asn Val Ala Asn Phe Gln Lys Ala Lys 100
105 110 gat gac gct gtt aag
caa tta act gga ggt att gag ctt ctg ttc aag 384Asp Asp Ala Val Lys
Gln Leu Thr Gly Gly Ile Glu Leu Leu Phe Lys 115
120 125 aaa aat aag gtc acc
tat tat aaa ggt aat ggt tca ttc gaa gac gaa 432Lys Asn Lys Val Thr
Tyr Tyr Lys Gly Asn Gly Ser Phe Glu Asp Glu 130
135 140 acg aag atc aga gta
act ccc gtt gat ggg ttg gaa ggc act gtc aag 480Thr Lys Ile Arg Val
Thr Pro Val Asp Gly Leu Glu Gly Thr Val Lys 145
150 155 160 gaa gac cac ata cta
gat gtt aag aac atc ata gtc gcc acg ggc tct 528Glu Asp His Ile Leu
Asp Val Lys Asn Ile Ile Val Ala Thr Gly Ser 165
170 175 gaa gtt aca ccc ttc
ccc ggt att gaa ata gat gag gaa aaa att gtc 576Glu Val Thr Pro Phe
Pro Gly Ile Glu Ile Asp Glu Glu Lys Ile Val 180
185 190 tct tca aca ggt gct
ctt tcg tta aag gaa att ccc aaa aga tta acc 624Ser Ser Thr Gly Ala
Leu Ser Leu Lys Glu Ile Pro Lys Arg Leu Thr 195
200 205 atc att ggt gga gga
atc atc gga ttg gaa atg ggt tca gtt tac tct 672Ile Ile Gly Gly Gly
Ile Ile Gly Leu Glu Met Gly Ser Val Tyr Ser 210
215 220 aga tta ggc tcc aag
gtt act gta gta gaa ttt caa cct caa att ggt 720Arg Leu Gly Ser Lys
Val Thr Val Val Glu Phe Gln Pro Gln Ile Gly 225
230 235 240 gca tct atg gac ggc
gag gtt gcc aaa gcc acc caa aag ttc ttg aaa 768Ala Ser Met Asp Gly
Glu Val Ala Lys Ala Thr Gln Lys Phe Leu Lys 245
250 255 aag caa ggt ttg gac
ttc aaa tta agc acc aaa gtt att tct gca aag 816Lys Gln Gly Leu Asp
Phe Lys Leu Ser Thr Lys Val Ile Ser Ala Lys 260
265 270 aga aac gac gac aag
aac gtc gtc gaa att gtt gta gaa gat act aaa 864Arg Asn Asp Asp Lys
Asn Val Val Glu Ile Val Val Glu Asp Thr Lys 275
280 285 acg aat aag caa gaa
aat ttg gaa gct gaa gtt ttg ctg gtt gct gtt 912Thr Asn Lys Gln Glu
Asn Leu Glu Ala Glu Val Leu Leu Val Ala Val 290
295 300 ggt aga aga cct tac
att gct ggc tta ggg gct gaa aag att gga tta 960Gly Arg Arg Pro Tyr
Ile Ala Gly Leu Gly Ala Glu Lys Ile Gly Leu 305
310 315 320 gaa gta gac aaa agg
gga cgc cta gtc att gat gac caa ttt aat tcc 1008Glu Val Asp Lys Arg
Gly Arg Leu Val Ile Asp Asp Gln Phe Asn Ser 325
330 335 aag ttc cca cac att
aaa gtg gta gga gat gtt aca ttt ggt cca atg 1056Lys Phe Pro His Ile
Lys Val Val Gly Asp Val Thr Phe Gly Pro Met 340
345 350 ctg gct cac aaa gcc
gaa gag gaa ggt att gca gct gtc gaa atg ttg 1104Leu Ala His Lys Ala
Glu Glu Glu Gly Ile Ala Ala Val Glu Met Leu 355
360 365 aaa act ggt cac ggt
cat gtc aac tat aac aac att cct tcg gtc atg 1152Lys Thr Gly His Gly
His Val Asn Tyr Asn Asn Ile Pro Ser Val Met 370
375 380 tat tct cac cca gaa
gta gca tgg gtt ggt aaa acc gaa gag caa ttg 1200Tyr Ser His Pro Glu
Val Ala Trp Val Gly Lys Thr Glu Glu Gln Leu 385
390 395 400 aaa gaa gcc ggc att
gac tat aaa att ggt aag ttc ccc ttt gcg gcc 1248Lys Glu Ala Gly Ile
Asp Tyr Lys Ile Gly Lys Phe Pro Phe Ala Ala 405
410 415 aat tca aga gcc aag
acc aac caa gac act gaa ggt ttc gtg aag att 1296Asn Ser Arg Ala Lys
Thr Asn Gln Asp Thr Glu Gly Phe Val Lys Ile 420
425 430 ttg atc gat tcc aag
acc gag cgt att ttg ggg gct cac att atc ggt 1344Leu Ile Asp Ser Lys
Thr Glu Arg Ile Leu Gly Ala His Ile Ile Gly 435
440 445 cca aat gcc ggt gaa
atg att gct gaa gct ggc tta gcc tta gaa tat 1392Pro Asn Ala Gly Glu
Met Ile Ala Glu Ala Gly Leu Ala Leu Glu Tyr 450
455 460 ggc gct tcc gca gaa
gat gtt gct agg gtc tgc cat gct cat cct act 1440Gly Ala Ser Ala Glu
Asp Val Ala Arg Val Cys His Ala His Pro Thr 465
470 475 480 ttg tcc gaa gca ttt
aag gaa gct aac atg gct gcc tat gat aaa gct 1488Leu Ser Glu Ala Phe
Lys Glu Ala Asn Met Ala Ala Tyr Asp Lys Ala 485
490 495 att cat tgt tga
1500Ile His Cys
108499PRT
Saccharomyces cerevisiae 108 Met Leu Arg Ile Arg Ser Leu Leu Asn Asn Lys
Arg Ala Phe Ser Ser 1 5 10
15 Thr Val Arg Thr Leu Thr Ile Asn Lys Ser His Asp Val Val Ile Ile
20 25 30 Gly Gly
Gly Pro Ala Gly Tyr Val Ala Ala Ile Lys Ala Ala Gln Leu 35
40 45 Gly Phe Asn Thr Ala Cys Val
Glu Lys Arg Gly Lys Leu Gly Gly Thr 50 55
60 Cys Leu Asn Val Gly Cys Ile Pro Ser Lys Ala Leu
Leu Asn Asn Ser 65 70 75
80 His Leu Phe His Gln Met His Thr Glu Ala Gln Lys Arg Gly Ile Asp
85 90 95 Val Asn Gly
Asp Ile Lys Ile Asn Val Ala Asn Phe Gln Lys Ala Lys 100
105 110 Asp Asp Ala Val Lys Gln Leu Thr
Gly Gly Ile Glu Leu Leu Phe Lys 115 120
125 Lys Asn Lys Val Thr Tyr Tyr Lys Gly Asn Gly Ser Phe
Glu Asp Glu 130 135 140
Thr Lys Ile Arg Val Thr Pro Val Asp Gly Leu Glu Gly Thr Val Lys 145
150 155 160 Glu Asp His Ile
Leu Asp Val Lys Asn Ile Ile Val Ala Thr Gly Ser 165
170 175 Glu Val Thr Pro Phe Pro Gly Ile Glu
Ile Asp Glu Glu Lys Ile Val 180 185
190 Ser Ser Thr Gly Ala Leu Ser Leu Lys Glu Ile Pro Lys Arg
Leu Thr 195 200 205
Ile Ile Gly Gly Gly Ile Ile Gly Leu Glu Met Gly Ser Val Tyr Ser 210
215 220 Arg Leu Gly Ser Lys
Val Thr Val Val Glu Phe Gln Pro Gln Ile Gly 225 230
235 240 Ala Ser Met Asp Gly Glu Val Ala Lys Ala
Thr Gln Lys Phe Leu Lys 245 250
255 Lys Gln Gly Leu Asp Phe Lys Leu Ser Thr Lys Val Ile Ser Ala
Lys 260 265 270 Arg
Asn Asp Asp Lys Asn Val Val Glu Ile Val Val Glu Asp Thr Lys 275
280 285 Thr Asn Lys Gln Glu Asn
Leu Glu Ala Glu Val Leu Leu Val Ala Val 290 295
300 Gly Arg Arg Pro Tyr Ile Ala Gly Leu Gly Ala
Glu Lys Ile Gly Leu 305 310 315
320 Glu Val Asp Lys Arg Gly Arg Leu Val Ile Asp Asp Gln Phe Asn Ser
325 330 335 Lys Phe
Pro His Ile Lys Val Val Gly Asp Val Thr Phe Gly Pro Met 340
345 350 Leu Ala His Lys Ala Glu Glu
Glu Gly Ile Ala Ala Val Glu Met Leu 355 360
365 Lys Thr Gly His Gly His Val Asn Tyr Asn Asn Ile
Pro Ser Val Met 370 375 380
Tyr Ser His Pro Glu Val Ala Trp Val Gly Lys Thr Glu Glu Gln Leu 385
390 395 400 Lys Glu Ala
Gly Ile Asp Tyr Lys Ile Gly Lys Phe Pro Phe Ala Ala 405
410 415 Asn Ser Arg Ala Lys Thr Asn Gln
Asp Thr Glu Gly Phe Val Lys Ile 420 425
430 Leu Ile Asp Ser Lys Thr Glu Arg Ile Leu Gly Ala His
Ile Ile Gly 435 440 445
Pro Asn Ala Gly Glu Met Ile Ala Glu Ala Gly Leu Ala Leu Glu Tyr 450
455 460 Gly Ala Ser Ala
Glu Asp Val Ala Arg Val Cys His Ala His Pro Thr 465 470
475 480 Leu Ser Glu Ala Phe Lys Glu Ala Asn
Met Ala Ala Tyr Asp Lys Ala 485 490
495 Ile His Cys 1091233DNASaccharomyces
cerevisiaeCDS(1)..(1233) 109atg cta agt gca att tcc aaa gtc tcc act tta
aaa tca tgt aca aga 48Met Leu Ser Ala Ile Ser Lys Val Ser Thr Leu
Lys Ser Cys Thr Arg 1 5 10
15 tat tta acc aaa tgc aac tat cat gca tca gct
aaa tta ctt gct gta 96Tyr Leu Thr Lys Cys Asn Tyr His Ala Ser Ala
Lys Leu Leu Ala Val 20 25
30 aag aca ttt tca atg cct gca atg tct cct act
atg gag aaa ggg ggg 144Lys Thr Phe Ser Met Pro Ala Met Ser Pro Thr
Met Glu Lys Gly Gly 35 40
45 att gtg tct tgg aaa tat aaa gtt ggc gaa cca
ttc agc gcg ggc gat 192Ile Val Ser Trp Lys Tyr Lys Val Gly Glu Pro
Phe Ser Ala Gly Asp 50 55
60 gtg ata tta gaa gtg gaa aca gat aaa tct caa
att gat gtg gaa gca 240Val Ile Leu Glu Val Glu Thr Asp Lys Ser Gln
Ile Asp Val Glu Ala 65 70 75
80 ctg gac gat ggt aaa cta gct aag atc ctg aaa
gat gaa ggc tct aaa 288Leu Asp Asp Gly Lys Leu Ala Lys Ile Leu Lys
Asp Glu Gly Ser Lys 85 90
95 gat gtt gat gtt ggt gaa cct att gct tat att
gct gat gtt gat gat 336Asp Val Asp Val Gly Glu Pro Ile Ala Tyr Ile
Ala Asp Val Asp Asp 100 105
110 gat tta gct act ata aag tta ccc caa gag gcc
aac acc gca aat gcg 384Asp Leu Ala Thr Ile Lys Leu Pro Gln Glu Ala
Asn Thr Ala Asn Ala 115 120
125 aaa tct att gaa att aag aag cca tcc gca gat
agt act gaa gca aca 432Lys Ser Ile Glu Ile Lys Lys Pro Ser Ala Asp
Ser Thr Glu Ala Thr 130 135
140 caa caa cat tta aaa aaa gcc aca gtt aca cca
ata aaa acc gtt gac 480Gln Gln His Leu Lys Lys Ala Thr Val Thr Pro
Ile Lys Thr Val Asp 145 150 155
160 ggc agc caa gcc aat ctt gaa cag acg cta tta
cca tcc gtg tca tta 528Gly Ser Gln Ala Asn Leu Glu Gln Thr Leu Leu
Pro Ser Val Ser Leu 165 170
175 cta ctg gct gag aac aat ata tcc aaa caa aag
gct ttg aag gaa att 576Leu Leu Ala Glu Asn Asn Ile Ser Lys Gln Lys
Ala Leu Lys Glu Ile 180 185
190 gcg cca tct ggt tcc aac ggt aga cta tta aag
ggt gat gtg cta gca 624Ala Pro Ser Gly Ser Asn Gly Arg Leu Leu Lys
Gly Asp Val Leu Ala 195 200
205 tac cta ggg aaa ata cca caa gat tcg gtt aac
aag gta aca gaa ttt 672Tyr Leu Gly Lys Ile Pro Gln Asp Ser Val Asn
Lys Val Thr Glu Phe 210 215
220 atc aag aag aac gaa cgt ctc gat tta tcg aac
att aaa cct ata cag 720Ile Lys Lys Asn Glu Arg Leu Asp Leu Ser Asn
Ile Lys Pro Ile Gln 225 230 235
240 ctc aaa cca aaa ata gcc gag caa gct caa aca
aaa gct gcc gac aag 768Leu Lys Pro Lys Ile Ala Glu Gln Ala Gln Thr
Lys Ala Ala Asp Lys 245 250
255 cca aag att act cct gta gaa ttt gaa gag caa
tta gtg ttc cat gct 816Pro Lys Ile Thr Pro Val Glu Phe Glu Glu Gln
Leu Val Phe His Ala 260 265
270 ccc gcc tct att ccg ttt gac aaa ctg agt gaa
tca ttg aac tct ttc 864Pro Ala Ser Ile Pro Phe Asp Lys Leu Ser Glu
Ser Leu Asn Ser Phe 275 280
285 atg aaa gaa gct tac cag ttc tca cac gga aca
cca cta atg gac aca 912Met Lys Glu Ala Tyr Gln Phe Ser His Gly Thr
Pro Leu Met Asp Thr 290 295
300 aat tcg aaa tac ttt gac cct att ttc gag gac
ctt gtc acc ttg agc 960Asn Ser Lys Tyr Phe Asp Pro Ile Phe Glu Asp
Leu Val Thr Leu Ser 305 310 315
320 cca aga gag cca aga ttt aaa ttt tcc tat gac
ttg atg caa att ccc 1008Pro Arg Glu Pro Arg Phe Lys Phe Ser Tyr Asp
Leu Met Gln Ile Pro 325 330
335 aaa gct aat aac atg caa gac acg tac ggt caa
gaa gac ata ttt gac 1056Lys Ala Asn Asn Met Gln Asp Thr Tyr Gly Gln
Glu Asp Ile Phe Asp 340 345
350 ctc tta aca ggt tca gac gcg act gcc tca tca
gta aga ccc gtt gaa 1104Leu Leu Thr Gly Ser Asp Ala Thr Ala Ser Ser
Val Arg Pro Val Glu 355 360
365 aag aac tta cct gaa aaa aac gaa tat ata cta
gcg ttg aat gtt agc 1152Lys Asn Leu Pro Glu Lys Asn Glu Tyr Ile Leu
Ala Leu Asn Val Ser 370 375
380 gtc aac aac aag aag ttt aat gac gcg gag gcc
aag gca aaa aga ttc 1200Val Asn Asn Lys Lys Phe Asn Asp Ala Glu Ala
Lys Ala Lys Arg Phe 385 390 395
400 ctt gat tac gta agg gag tta gaa tca ttt tga
1233Leu Asp Tyr Val Arg Glu Leu Glu Ser Phe
405 410
110410PRTSaccharomyces cerevisiae 110Met Leu
Ser Ala Ile Ser Lys Val Ser Thr Leu Lys Ser Cys Thr Arg 1 5
10 15 Tyr Leu Thr Lys Cys Asn Tyr
His Ala Ser Ala Lys Leu Leu Ala Val 20 25
30 Lys Thr Phe Ser Met Pro Ala Met Ser Pro Thr Met
Glu Lys Gly Gly 35 40 45
Ile Val Ser Trp Lys Tyr Lys Val Gly Glu Pro Phe Ser Ala Gly Asp
50 55 60 Val Ile Leu
Glu Val Glu Thr Asp Lys Ser Gln Ile Asp Val Glu Ala 65
70 75 80 Leu Asp Asp Gly Lys Leu Ala
Lys Ile Leu Lys Asp Glu Gly Ser Lys 85
90 95 Asp Val Asp Val Gly Glu Pro Ile Ala Tyr Ile
Ala Asp Val Asp Asp 100 105
110 Asp Leu Ala Thr Ile Lys Leu Pro Gln Glu Ala Asn Thr Ala Asn
Ala 115 120 125 Lys
Ser Ile Glu Ile Lys Lys Pro Ser Ala Asp Ser Thr Glu Ala Thr 130
135 140 Gln Gln His Leu Lys Lys
Ala Thr Val Thr Pro Ile Lys Thr Val Asp 145 150
155 160 Gly Ser Gln Ala Asn Leu Glu Gln Thr Leu Leu
Pro Ser Val Ser Leu 165 170
175 Leu Leu Ala Glu Asn Asn Ile Ser Lys Gln Lys Ala Leu Lys Glu Ile
180 185 190 Ala Pro
Ser Gly Ser Asn Gly Arg Leu Leu Lys Gly Asp Val Leu Ala 195
200 205 Tyr Leu Gly Lys Ile Pro Gln
Asp Ser Val Asn Lys Val Thr Glu Phe 210 215
220 Ile Lys Lys Asn Glu Arg Leu Asp Leu Ser Asn Ile
Lys Pro Ile Gln 225 230 235
240 Leu Lys Pro Lys Ile Ala Glu Gln Ala Gln Thr Lys Ala Ala Asp Lys
245 250 255 Pro Lys Ile
Thr Pro Val Glu Phe Glu Glu Gln Leu Val Phe His Ala 260
265 270 Pro Ala Ser Ile Pro Phe Asp Lys
Leu Ser Glu Ser Leu Asn Ser Phe 275 280
285 Met Lys Glu Ala Tyr Gln Phe Ser His Gly Thr Pro Leu
Met Asp Thr 290 295 300
Asn Ser Lys Tyr Phe Asp Pro Ile Phe Glu Asp Leu Val Thr Leu Ser 305
310 315 320 Pro Arg Glu Pro
Arg Phe Lys Phe Ser Tyr Asp Leu Met Gln Ile Pro 325
330 335 Lys Ala Asn Asn Met Gln Asp Thr Tyr
Gly Gln Glu Asp Ile Phe Asp 340 345
350 Leu Leu Thr Gly Ser Asp Ala Thr Ala Ser Ser Val Arg Pro
Val Glu 355 360 365
Lys Asn Leu Pro Glu Lys Asn Glu Tyr Ile Leu Ala Leu Asn Val Ser 370
375 380 Val Asn Asn Lys Lys
Phe Asn Asp Ala Glu Ala Lys Ala Lys Arg Phe 385 390
395 400 Leu Asp Tyr Val Arg Glu Leu Glu Ser Phe
405 410 1111692DNASaccharomyces
cerevisiaeCDS(1)..(1692) 111atg tct gaa att act ttg ggt aaa tat ttg ttc
gaa aga tta aag caa 48Met Ser Glu Ile Thr Leu Gly Lys Tyr Leu Phe
Glu Arg Leu Lys Gln 1 5 10
15 gtc aac gtt aac acc gtt ttc ggt ttg cca ggt
gac ttc aac ttg tcc 96Val Asn Val Asn Thr Val Phe Gly Leu Pro Gly
Asp Phe Asn Leu Ser 20 25
30 ttg ttg gac aag atc tac gaa gtt gaa ggt atg
aga tgg gct ggt aac 144Leu Leu Asp Lys Ile Tyr Glu Val Glu Gly Met
Arg Trp Ala Gly Asn 35 40
45 gcc aac gaa ttg aac gct gct tac gcc gct gat
ggt tac gct cgt atc 192Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp
Gly Tyr Ala Arg Ile 50 55
60 aag ggt atg tct tgt atc atc acc acc ttc ggt
gtc ggt gaa ttg tct 240Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly
Val Gly Glu Leu Ser 65 70 75
80 gct ttg aac ggt att gcc ggt tct tac gct gaa
cac gtc ggt gtt ttg 288Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu
His Val Gly Val Leu 85 90
95 cac gtt gtt ggt gtc cca tcc atc tct gct caa
gct aag caa ttg ttg 336His Val Val Gly Val Pro Ser Ile Ser Ala Gln
Ala Lys Gln Leu Leu 100 105
110 ttg cac cac acc ttg ggt aac ggt gac ttc act
gtt ttc cac aga atg 384Leu His His Thr Leu Gly Asn Gly Asp Phe Thr
Val Phe His Arg Met 115 120
125 tct gcc aac att tct gaa acc act gct atg atc
act gac att gct acc 432Ser Ala Asn Ile Ser Glu Thr Thr Ala Met Ile
Thr Asp Ile Ala Thr 130 135
140 gcc cca gct gaa att gac aga tgt atc aga acc
act tac gtc acc caa 480Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr
Thr Tyr Val Thr Gln 145 150 155
160 aga cca gtc tac tta ggt ttg cca gct aac ttg
gtc gac ttg aac gtc 528Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu
Val Asp Leu Asn Val 165 170
175 cca gct aag ttg ttg caa act cca att gac atg
tct ttg aag cca aac 576Pro Ala Lys Leu Leu Gln Thr Pro Ile Asp Met
Ser Leu Lys Pro Asn 180 185
190 gat gct gaa tcc gaa aag gaa gtc att gac acc
atc ttg gct ttg gtc 624Asp Ala Glu Ser Glu Lys Glu Val Ile Asp Thr
Ile Leu Ala Leu Val 195 200
205 aag gat gct aag aac cca gtt atc ttg gct gat
gct tgt tgt tcc aga 672Lys Asp Ala Lys Asn Pro Val Ile Leu Ala Asp
Ala Cys Cys Ser Arg 210 215
220 cac gac gtc aag gct gaa act aag aag ttg att
gac ttg act caa ttc 720His Asp Val Lys Ala Glu Thr Lys Lys Leu Ile
Asp Leu Thr Gln Phe 225 230 235
240 cca gct ttc gtc acc cca atg ggt aag ggt tcc
att gac gaa caa cac 768Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser
Ile Asp Glu Gln His 245 250
255 cca aga tac ggt ggt gtt tac gtc ggt acc ttg
tcc aag cca gaa gtt 816Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu
Ser Lys Pro Glu Val 260 265
270 aag gaa gcc gtt gaa tct gct gac ttg att ttg
tct gtc ggt gct ttg 864Lys Glu Ala Val Glu Ser Ala Asp Leu Ile Leu
Ser Val Gly Ala Leu 275 280
285 ttg tct gat ttc aac acc ggt tct ttc tct tac
tct tac aag acc aag 912Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr
Ser Tyr Lys Thr Lys 290 295
300 aac att gtc gaa ttc cac tcc gac cac atg aag
atc aga aac gcc act 960Asn Ile Val Glu Phe His Ser Asp His Met Lys
Ile Arg Asn Ala Thr 305 310 315
320 ttc cca ggt gtc caa atg aaa ttc gtt ttg caa
aag ttg ttg acc act 1008Phe Pro Gly Val Gln Met Lys Phe Val Leu Gln
Lys Leu Leu Thr Thr 325 330
335 att gct gac gcc gct aag ggt tac aag cca gtt
gct gtc cca gct aga 1056Ile Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val
Ala Val Pro Ala Arg 340 345
350 act cca gct aac gct gct gtc cca gct tct acc
cca ttg aag caa gaa 1104Thr Pro Ala Asn Ala Ala Val Pro Ala Ser Thr
Pro Leu Lys Gln Glu 355 360
365 tgg atg tgg aac caa ttg ggt aac ttc ttg caa
gaa ggt gat gtt gtc 1152Trp Met Trp Asn Gln Leu Gly Asn Phe Leu Gln
Glu Gly Asp Val Val 370 375
380 att gct gaa acc ggt acc tcc gct ttc ggt atc
aac caa acc act ttc 1200Ile Ala Glu Thr Gly Thr Ser Ala Phe Gly Ile
Asn Gln Thr Thr Phe 385 390 395
400 cca aac aac acc tac ggt atc tct caa gtc tta
tgg ggt tcc att ggt 1248Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val Leu
Trp Gly Ser Ile Gly 405 410
415 ttc acc act ggt gct acc ttg ggt gct gct ttc
gct gct gaa gaa att 1296Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe
Ala Ala Glu Glu Ile 420 425
430 gat cca aag aag aga gtt atc tta ttc att ggt
gac ggt tct ttg caa 1344Asp Pro Lys Lys Arg Val Ile Leu Phe Ile Gly
Asp Gly Ser Leu Gln 435 440
445 ttg act gtt caa gaa atc tcc acc atg atc aga
tgg ggc ttg aag cca 1392Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg
Trp Gly Leu Lys Pro 450 455
460 tac ttg ttc gtc ttg aac aac gat ggt tac acc
att gaa aag ttg att 1440Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr
Ile Glu Lys Leu Ile 465 470 475
480 cac ggt cca aag gct caa tac aac gaa att caa
ggt tgg gac cac cta 1488His Gly Pro Lys Ala Gln Tyr Asn Glu Ile Gln
Gly Trp Asp His Leu 485 490
495 tcc ttg ttg cca act ttc ggt gct aag gac tat
gaa acc cac aga gtc 1536Ser Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr
Glu Thr His Arg Val 500 505
510 gct acc acc ggt gaa tgg gac aag ttg acc caa
gac aag tct ttc aac 1584Ala Thr Thr Gly Glu Trp Asp Lys Leu Thr Gln
Asp Lys Ser Phe Asn 515 520
525 gac aac tct aag atc aga atg att gaa atc atg
ttg cca gtc ttc gat 1632Asp Asn Ser Lys Ile Arg Met Ile Glu Ile Met
Leu Pro Val Phe Asp 530 535
540 gct cca caa aac ttg gtt gaa caa gct aag ttg
act gct gct acc aac 1680Ala Pro Gln Asn Leu Val Glu Gln Ala Lys Leu
Thr Ala Ala Thr Asn 545 550 555
560 gct aag caa taa
1692Ala Lys Gln
112563PRTSaccharomyces cerevisiae 112Met Ser
Glu Ile Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys Gln 1 5
10 15 Val Asn Val Asn Thr Val Phe
Gly Leu Pro Gly Asp Phe Asn Leu Ser 20 25
30 Leu Leu Asp Lys Ile Tyr Glu Val Glu Gly Met Arg
Trp Ala Gly Asn 35 40 45
Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile
50 55 60 Lys Gly Met
Ser Cys Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser 65
70 75 80 Ala Leu Asn Gly Ile Ala Gly
Ser Tyr Ala Glu His Val Gly Val Leu 85
90 95 His Val Val Gly Val Pro Ser Ile Ser Ala Gln
Ala Lys Gln Leu Leu 100 105
110 Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg
Met 115 120 125 Ser
Ala Asn Ile Ser Glu Thr Thr Ala Met Ile Thr Asp Ile Ala Thr 130
135 140 Ala Pro Ala Glu Ile Asp
Arg Cys Ile Arg Thr Thr Tyr Val Thr Gln 145 150
155 160 Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu
Val Asp Leu Asn Val 165 170
175 Pro Ala Lys Leu Leu Gln Thr Pro Ile Asp Met Ser Leu Lys Pro Asn
180 185 190 Asp Ala
Glu Ser Glu Lys Glu Val Ile Asp Thr Ile Leu Ala Leu Val 195
200 205 Lys Asp Ala Lys Asn Pro Val
Ile Leu Ala Asp Ala Cys Cys Ser Arg 210 215
220 His Asp Val Lys Ala Glu Thr Lys Lys Leu Ile Asp
Leu Thr Gln Phe 225 230 235
240 Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser Ile Asp Glu Gln His
245 250 255 Pro Arg Tyr
Gly Gly Val Tyr Val Gly Thr Leu Ser Lys Pro Glu Val 260
265 270 Lys Glu Ala Val Glu Ser Ala Asp
Leu Ile Leu Ser Val Gly Ala Leu 275 280
285 Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr
Lys Thr Lys 290 295 300
Asn Ile Val Glu Phe His Ser Asp His Met Lys Ile Arg Asn Ala Thr 305
310 315 320 Phe Pro Gly Val
Gln Met Lys Phe Val Leu Gln Lys Leu Leu Thr Thr 325
330 335 Ile Ala Asp Ala Ala Lys Gly Tyr Lys
Pro Val Ala Val Pro Ala Arg 340 345
350 Thr Pro Ala Asn Ala Ala Val Pro Ala Ser Thr Pro Leu Lys
Gln Glu 355 360 365
Trp Met Trp Asn Gln Leu Gly Asn Phe Leu Gln Glu Gly Asp Val Val 370
375 380 Ile Ala Glu Thr Gly
Thr Ser Ala Phe Gly Ile Asn Gln Thr Thr Phe 385 390
395 400 Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val
Leu Trp Gly Ser Ile Gly 405 410
415 Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu
Ile 420 425 430 Asp
Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 435
440 445 Leu Thr Val Gln Glu Ile
Ser Thr Met Ile Arg Trp Gly Leu Lys Pro 450 455
460 Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr
Ile Glu Lys Leu Ile 465 470 475
480 His Gly Pro Lys Ala Gln Tyr Asn Glu Ile Gln Gly Trp Asp His Leu
485 490 495 Ser Leu
Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Thr His Arg Val 500
505 510 Ala Thr Thr Gly Glu Trp Asp
Lys Leu Thr Gln Asp Lys Ser Phe Asn 515 520
525 Asp Asn Ser Lys Ile Arg Met Ile Glu Ile Met Leu
Pro Val Phe Asp 530 535 540
Ala Pro Gln Asn Leu Val Glu Gln Ala Lys Leu Thr Ala Ala Thr Asn 545
550 555 560 Ala Lys Gln
1131692DNASaccharomyces cerevisiaeeCDS(1)..(1692) 113atg tct gaa ata acc
tta ggt aaa tat tta ttt gaa aga ttg agc caa 48Met Ser Glu Ile Thr
Leu Gly Lys Tyr Leu Phe Glu Arg Leu Ser Gln 1 5
10 15 gtc aac tgt aac acc
gtc ttc ggt ttg cca ggt gac ttt aac ttg tct 96Val Asn Cys Asn Thr
Val Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser 20
25 30 ctt ttg gat aag ctt
tat gaa gtc aaa ggt atg aga tgg gct ggt aac 144Leu Leu Asp Lys Leu
Tyr Glu Val Lys Gly Met Arg Trp Ala Gly Asn 35
40 45 gct aac gaa ttg aac
gct gcc tat gct gct gat ggt tac gct cgt atc 192Ala Asn Glu Leu Asn
Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile 50
55 60 aag ggt atg tcc tgt
att att acc acc ttc ggt gtt ggt gaa ttg tct 240Lys Gly Met Ser Cys
Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser 65
70 75 80 gct ttg aat ggt att
gcc ggt tct tac gct gaa cat gtc ggt gtt ttg 288Ala Leu Asn Gly Ile
Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu 85
90 95 cac gtt gtt ggt gtt
cca tcc atc tct tct caa gct aag caa ttg ttg 336His Val Val Gly Val
Pro Ser Ile Ser Ser Gln Ala Lys Gln Leu Leu 100
105 110 ttg cat cat acc ttg
ggt aac ggt gac ttc act gtt ttc cac aga atg 384Leu His His Thr Leu
Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 115
120 125 tct gcc aac att tct
gaa acc act gcc atg atc act gat att gct aac 432Ser Ala Asn Ile Ser
Glu Thr Thr Ala Met Ile Thr Asp Ile Ala Asn 130
135 140 gct cca gct gaa att
gac aga tgt atc aga acc acc tac act acc caa 480Ala Pro Ala Glu Ile
Asp Arg Cys Ile Arg Thr Thr Tyr Thr Thr Gln 145
150 155 160 aga cca gtc tac ttg
ggt ttg cca gct aac ttg gtt gac ttg aac gtc 528Arg Pro Val Tyr Leu
Gly Leu Pro Ala Asn Leu Val Asp Leu Asn Val 165
170 175 cca gcc aag tta ttg
gaa act cca att gac ttg tct ttg aag cca aac 576Pro Ala Lys Leu Leu
Glu Thr Pro Ile Asp Leu Ser Leu Lys Pro Asn 180
185 190 gac gct gaa gct gaa
gct gaa gtt gtt aga act gtt gtt gaa ttg atc 624Asp Ala Glu Ala Glu
Ala Glu Val Val Arg Thr Val Val Glu Leu Ile 195
200 205 aag gat gct aag aac
cca gtt atc ttg gct gat gct tgt gct tct aga 672Lys Asp Ala Lys Asn
Pro Val Ile Leu Ala Asp Ala Cys Ala Ser Arg 210
215 220 cat gat gtc aag gct
gaa act aag aag ttg atg gac ttg act caa ttc 720His Asp Val Lys Ala
Glu Thr Lys Lys Leu Met Asp Leu Thr Gln Phe 225
230 235 240 cca gtt tac gtc acc
cca atg ggt aag ggt gct att gac gaa caa cac 768Pro Val Tyr Val Thr
Pro Met Gly Lys Gly Ala Ile Asp Glu Gln His 245
250 255 cca aga tac ggt ggt
gtt tac gtt ggt acc ttg tct aga cca gaa gtt 816Pro Arg Tyr Gly Gly
Val Tyr Val Gly Thr Leu Ser Arg Pro Glu Val 260
265 270 aag aag gct gta gaa
tct gct gat ttg ata ttg tct atc ggt gct ttg 864Lys Lys Ala Val Glu
Ser Ala Asp Leu Ile Leu Ser Ile Gly Ala Leu 275
280 285 ttg tct gat ttc aat
acc ggt tct ttc tct tac tcc tac aag acc aaa 912Leu Ser Asp Phe Asn
Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290
295 300 aat atc gtt gaa ttc
cac tct gac cac atc aag atc aga aac gcc acc 960Asn Ile Val Glu Phe
His Ser Asp His Ile Lys Ile Arg Asn Ala Thr 305
310 315 320 ttc cca ggt gtt caa
atg aaa ttt gcc ttg caa aaa ttg ttg gat gct 1008Phe Pro Gly Val Gln
Met Lys Phe Ala Leu Gln Lys Leu Leu Asp Ala 325
330 335 att cca gaa gtc gtc
aag gac tac aaa cct gtt gct gtc cca gct aga 1056Ile Pro Glu Val Val
Lys Asp Tyr Lys Pro Val Ala Val Pro Ala Arg 340
345 350 gtt cca att acc aag
tct act cca gct aac act cca atg aag caa gaa 1104Val Pro Ile Thr Lys
Ser Thr Pro Ala Asn Thr Pro Met Lys Gln Glu 355
360 365 tgg atg tgg aac cat
ttg ggt aac ttc ttg aga gaa ggt gat att gtt 1152Trp Met Trp Asn His
Leu Gly Asn Phe Leu Arg Glu Gly Asp Ile Val 370
375 380 att gct gaa acc ggt
act tcc gcc ttc ggt att aac caa act act ttc 1200Ile Ala Glu Thr Gly
Thr Ser Ala Phe Gly Ile Asn Gln Thr Thr Phe 385
390 395 400 cca aca gat gta tac
gct atc gtc caa gtc ttg tgg ggt tcc att ggt 1248Pro Thr Asp Val Tyr
Ala Ile Val Gln Val Leu Trp Gly Ser Ile Gly 405
410 415 ttc aca gtc ggc gct
cta ttg ggt gct act atg gcc gct gaa gaa ctt 1296Phe Thr Val Gly Ala
Leu Leu Gly Ala Thr Met Ala Ala Glu Glu Leu 420
425 430 gat cca aag aag aga
gtt att tta ttc att ggt gac ggt tct cta caa 1344Asp Pro Lys Lys Arg
Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 435
440 445 ttg act gtt caa gaa
atc tct acc atg att aga tgg ggt ttg aag cca 1392Leu Thr Val Gln Glu
Ile Ser Thr Met Ile Arg Trp Gly Leu Lys Pro 450
455 460 tac att ttt gtc ttg
aat aac aac ggt tac acc att gaa aaa ttg att 1440Tyr Ile Phe Val Leu
Asn Asn Asn Gly Tyr Thr Ile Glu Lys Leu Ile 465
470 475 480 cac ggt cct cat gcc
gaa tat aat gaa att caa ggt tgg gac cac ttg 1488His Gly Pro His Ala
Glu Tyr Asn Glu Ile Gln Gly Trp Asp His Leu 485
490 495 gcc tta ttg cca act
ttt ggt gct aga aac tac gaa acc cac aga gtt 1536Ala Leu Leu Pro Thr
Phe Gly Ala Arg Asn Tyr Glu Thr His Arg Val 500
505 510 gct acc act ggt gaa
tgg gaa aag ttg act caa gac aag gac ttc caa 1584Ala Thr Thr Gly Glu
Trp Glu Lys Leu Thr Gln Asp Lys Asp Phe Gln 515
520 525 gac aac tct aag att
aga atg att gaa gtt atg ttg cca gtc ttt gat 1632Asp Asn Ser Lys Ile
Arg Met Ile Glu Val Met Leu Pro Val Phe Asp 530
535 540 gct cca caa aac ttg
gtt aaa caa gct caa ttg act gcc gct act aac 1680Ala Pro Gln Asn Leu
Val Lys Gln Ala Gln Leu Thr Ala Ala Thr Asn 545
550 555 560 gct aaa caa taa
1692Ala Lys Gln
114563PRTSaccharomyces cerevisiaee 114Met Ser Glu Ile Thr Leu Gly Lys Tyr
Leu Phe Glu Arg Leu Ser Gln 1 5 10
15 Val Asn Cys Asn Thr Val Phe Gly Leu Pro Gly Asp Phe Asn
Leu Ser 20 25 30
Leu Leu Asp Lys Leu Tyr Glu Val Lys Gly Met Arg Trp Ala Gly Asn
35 40 45 Ala Asn Glu Leu
Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile 50
55 60 Lys Gly Met Ser Cys Ile Ile Thr
Thr Phe Gly Val Gly Glu Leu Ser 65 70
75 80 Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His
Val Gly Val Leu 85 90
95 His Val Val Gly Val Pro Ser Ile Ser Ser Gln Ala Lys Gln Leu Leu
100 105 110 Leu His His
Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 115
120 125 Ser Ala Asn Ile Ser Glu Thr Thr
Ala Met Ile Thr Asp Ile Ala Asn 130 135
140 Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr Thr Tyr
Thr Thr Gln 145 150 155
160 Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Asn Val
165 170 175 Pro Ala Lys Leu
Leu Glu Thr Pro Ile Asp Leu Ser Leu Lys Pro Asn 180
185 190 Asp Ala Glu Ala Glu Ala Glu Val Val
Arg Thr Val Val Glu Leu Ile 195 200
205 Lys Asp Ala Lys Asn Pro Val Ile Leu Ala Asp Ala Cys Ala
Ser Arg 210 215 220
His Asp Val Lys Ala Glu Thr Lys Lys Leu Met Asp Leu Thr Gln Phe 225
230 235 240 Pro Val Tyr Val Thr
Pro Met Gly Lys Gly Ala Ile Asp Glu Gln His 245
250 255 Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr
Leu Ser Arg Pro Glu Val 260 265
270 Lys Lys Ala Val Glu Ser Ala Asp Leu Ile Leu Ser Ile Gly Ala
Leu 275 280 285 Leu
Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290
295 300 Asn Ile Val Glu Phe His
Ser Asp His Ile Lys Ile Arg Asn Ala Thr 305 310
315 320 Phe Pro Gly Val Gln Met Lys Phe Ala Leu Gln
Lys Leu Leu Asp Ala 325 330
335 Ile Pro Glu Val Val Lys Asp Tyr Lys Pro Val Ala Val Pro Ala Arg
340 345 350 Val Pro
Ile Thr Lys Ser Thr Pro Ala Asn Thr Pro Met Lys Gln Glu 355
360 365 Trp Met Trp Asn His Leu Gly
Asn Phe Leu Arg Glu Gly Asp Ile Val 370 375
380 Ile Ala Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn
Gln Thr Thr Phe 385 390 395
400 Pro Thr Asp Val Tyr Ala Ile Val Gln Val Leu Trp Gly Ser Ile Gly
405 410 415 Phe Thr Val
Gly Ala Leu Leu Gly Ala Thr Met Ala Ala Glu Glu Leu 420
425 430 Asp Pro Lys Lys Arg Val Ile Leu
Phe Ile Gly Asp Gly Ser Leu Gln 435 440
445 Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp Gly
Leu Lys Pro 450 455 460
Tyr Ile Phe Val Leu Asn Asn Asn Gly Tyr Thr Ile Glu Lys Leu Ile 465
470 475 480 His Gly Pro His
Ala Glu Tyr Asn Glu Ile Gln Gly Trp Asp His Leu 485
490 495 Ala Leu Leu Pro Thr Phe Gly Ala Arg
Asn Tyr Glu Thr His Arg Val 500 505
510 Ala Thr Thr Gly Glu Trp Glu Lys Leu Thr Gln Asp Lys Asp
Phe Gln 515 520 525
Asp Asn Ser Lys Ile Arg Met Ile Glu Val Met Leu Pro Val Phe Asp 530
535 540 Ala Pro Gln Asn Leu
Val Lys Gln Ala Gln Leu Thr Ala Ala Thr Asn 545 550
555 560 Ala Lys Gln 1151599DNASaccharomyces
cerevisiaeCDS(1)..(1599) 115atg tct gaa att act ctt gga aaa tac tta ttt
gaa aga ttg aag caa 48Met Ser Glu Ile Thr Leu Gly Lys Tyr Leu Phe
Glu Arg Leu Lys Gln 1 5 10
15 gtt aat gtt aac acc att ttt ggg cta cca ggc
gac ttc aac ttg tcc 96Val Asn Val Asn Thr Ile Phe Gly Leu Pro Gly
Asp Phe Asn Leu Ser 20 25
30 cta ttg gac aag att tac gag gta gat gga ttg
aga tgg gct ggt aat 144Leu Leu Asp Lys Ile Tyr Glu Val Asp Gly Leu
Arg Trp Ala Gly Asn 35 40
45 gca aat gag ctg aac gcc gcc tat gcc gcc gat
ggt tac gca cgc atc 192Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp
Gly Tyr Ala Arg Ile 50 55
60 aag ggt tta tct gtg ctg gta act act ttt ggc
gta ggt gaa tta tcc 240Lys Gly Leu Ser Val Leu Val Thr Thr Phe Gly
Val Gly Glu Leu Ser 65 70 75
80 gcc ttg aat ggt att gca gga tcg tat gca gaa
cac gtc ggt gta ctg 288Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu
His Val Gly Val Leu 85 90
95 cat gtt gtt ggt gtc ccc tct atc tcc gct cag
gct aag caa ttg ttg 336His Val Val Gly Val Pro Ser Ile Ser Ala Gln
Ala Lys Gln Leu Leu 100 105
110 ttg cat cat acc ttg ggt aac ggt gat ttt acc
gtt ttt cac aga atg 384Leu His His Thr Leu Gly Asn Gly Asp Phe Thr
Val Phe His Arg Met 115 120
125 tcc gcc aat atc tca gaa act aca tca atg att
aca gac att gct aca 432Ser Ala Asn Ile Ser Glu Thr Thr Ser Met Ile
Thr Asp Ile Ala Thr 130 135
140 gcc cct tca gaa atc gat agg ttg atc agg aca
aca ttt ata aca caa 480Ala Pro Ser Glu Ile Asp Arg Leu Ile Arg Thr
Thr Phe Ile Thr Gln 145 150 155
160 agg cct agc tac ttg ggg ttg cca gcg aat ttg
gta gat cta aag gtt 528Arg Pro Ser Tyr Leu Gly Leu Pro Ala Asn Leu
Val Asp Leu Lys Val 165 170
175 cct ggt tct ctt ttg gaa aaa ccg att gat cta
tca tta aaa cct aac 576Pro Gly Ser Leu Leu Glu Lys Pro Ile Asp Leu
Ser Leu Lys Pro Asn 180 185
190 gat ccc gaa gct gaa aag gaa gtt att gat acc
gta cta gaa ttg atc 624Asp Pro Glu Ala Glu Lys Glu Val Ile Asp Thr
Val Leu Glu Leu Ile 195 200
205 cag aat tcg aaa aac cct gtt ata cta tcg gat
gcc tgt gct tct agg 672Gln Asn Ser Lys Asn Pro Val Ile Leu Ser Asp
Ala Cys Ala Ser Arg 210 215
220 cac aac gtt aaa aaa gaa acc cag aag tta att
gat ttg acg caa ttc 720His Asn Val Lys Lys Glu Thr Gln Lys Leu Ile
Asp Leu Thr Gln Phe 225 230 235
240 cca gct ttt gtg aca cct cta ggt aaa ggg tca
ata gat gaa cag cat 768Pro Ala Phe Val Thr Pro Leu Gly Lys Gly Ser
Ile Asp Glu Gln His 245 250
255 ccc aga tat ggc ggt gtt tat gtg gga acg ctg
tcc aaa caa gac gtg 816Pro Arg Tyr Gly Gly Val Tyr Val Gly Thr Leu
Ser Lys Gln Asp Val 260 265
270 aaa cag gcc gtt gag tcg gct gat ttg atc ctt
tcg gtc ggt gct ttg 864Lys Gln Ala Val Glu Ser Ala Asp Leu Ile Leu
Ser Val Gly Ala Leu 275 280
285 ctc tct gat ttt aac aca ggt tcg ttt tcc tac
tcc tac aag act aaa 912Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr
Ser Tyr Lys Thr Lys 290 295
300 aat gta gtg gag ttt cat tcc gat tac gta aag
gtg aag aac gct acg 960Asn Val Val Glu Phe His Ser Asp Tyr Val Lys
Val Lys Asn Ala Thr 305 310 315
320 ttc ctc ggt gta caa atg aaa ttt gca cta caa
aac tta ctg aag gtt 1008Phe Leu Gly Val Gln Met Lys Phe Ala Leu Gln
Asn Leu Leu Lys Val 325 330
335 att ccc gat gtt gtt aag ggc tac aag agc gtt
ccc gta cca acc aaa 1056Ile Pro Asp Val Val Lys Gly Tyr Lys Ser Val
Pro Val Pro Thr Lys 340 345
350 act ccc gca aac aaa ggt gta cct gct agc acg
ccc ttg aaa caa gag 1104Thr Pro Ala Asn Lys Gly Val Pro Ala Ser Thr
Pro Leu Lys Gln Glu 355 360
365 tgg ttg tgg aac gaa ttg tcc aaa ttc ttg caa
gaa ggt gat gtt atc 1152Trp Leu Trp Asn Glu Leu Ser Lys Phe Leu Gln
Glu Gly Asp Val Ile 370 375
380 att tcc gag acc ggc acg tct gcc ttc ggt atc
aat caa act atc ttt 1200Ile Ser Glu Thr Gly Thr Ser Ala Phe Gly Ile
Asn Gln Thr Ile Phe 385 390 395
400 cct aag gac gcc tac ggt atc tcg cag gtg ttg
tgg ggg tcc atc ggt 1248Pro Lys Asp Ala Tyr Gly Ile Ser Gln Val Leu
Trp Gly Ser Ile Gly 405 410
415 ttt aca aca gga gca act tta ggt gct gcc ttt
gcc gct gag gag att 1296Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe
Ala Ala Glu Glu Ile 420 425
430 gac ccc aac aag aga gtc atc tta ttc ata ggt
gac ggg tct ttg cag 1344Asp Pro Asn Lys Arg Val Ile Leu Phe Ile Gly
Asp Gly Ser Leu Gln 435 440
445 tta acc gtc caa gaa atc tcc acc atg atc aga
tgg ggg tta aag ccg 1392Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg
Trp Gly Leu Lys Pro 450 455
460 tat ctt ttt gtc ctt aac aac gac ggc tac act
atc gaa aag ctg att 1440Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr
Ile Glu Lys Leu Ile 465 470 475
480 cat ggg cct cac gca gag tac aac gaa atc cag
acc tgg gat cac ctc 1488His Gly Pro His Ala Glu Tyr Asn Glu Ile Gln
Thr Trp Asp His Leu 485 490
495 gcc ctg ttg ccc gca ttt ggt gcg aaa aag tac
gaa aat cac aag atc 1536Ala Leu Leu Pro Ala Phe Gly Ala Lys Lys Tyr
Glu Asn His Lys Ile 500 505
510 gcc act acg ggt gag tgg gat gcc tta acc act
gat tca gag ttc cag 1584Ala Thr Thr Gly Glu Trp Asp Ala Leu Thr Thr
Asp Ser Glu Phe Gln 515 520
525 aaa aac tcg gtg atc
1599Lys Asn Ser Val Ile
530
116533PRTSaccharomyces cerevisiae 116Met Ser
Glu Ile Thr Leu Gly Lys Tyr Leu Phe Glu Arg Leu Lys Gln 1 5
10 15 Val Asn Val Asn Thr Ile Phe
Gly Leu Pro Gly Asp Phe Asn Leu Ser 20 25
30 Leu Leu Asp Lys Ile Tyr Glu Val Asp Gly Leu Arg
Trp Ala Gly Asn 35 40 45
Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Ile
50 55 60 Lys Gly Leu
Ser Val Leu Val Thr Thr Phe Gly Val Gly Glu Leu Ser 65
70 75 80 Ala Leu Asn Gly Ile Ala Gly
Ser Tyr Ala Glu His Val Gly Val Leu 85
90 95 His Val Val Gly Val Pro Ser Ile Ser Ala Gln
Ala Lys Gln Leu Leu 100 105
110 Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg
Met 115 120 125 Ser
Ala Asn Ile Ser Glu Thr Thr Ser Met Ile Thr Asp Ile Ala Thr 130
135 140 Ala Pro Ser Glu Ile Asp
Arg Leu Ile Arg Thr Thr Phe Ile Thr Gln 145 150
155 160 Arg Pro Ser Tyr Leu Gly Leu Pro Ala Asn Leu
Val Asp Leu Lys Val 165 170
175 Pro Gly Ser Leu Leu Glu Lys Pro Ile Asp Leu Ser Leu Lys Pro Asn
180 185 190 Asp Pro
Glu Ala Glu Lys Glu Val Ile Asp Thr Val Leu Glu Leu Ile 195
200 205 Gln Asn Ser Lys Asn Pro Val
Ile Leu Ser Asp Ala Cys Ala Ser Arg 210 215
220 His Asn Val Lys Lys Glu Thr Gln Lys Leu Ile Asp
Leu Thr Gln Phe 225 230 235
240 Pro Ala Phe Val Thr Pro Leu Gly Lys Gly Ser Ile Asp Glu Gln His
245 250 255 Pro Arg Tyr
Gly Gly Val Tyr Val Gly Thr Leu Ser Lys Gln Asp Val 260
265 270 Lys Gln Ala Val Glu Ser Ala Asp
Leu Ile Leu Ser Val Gly Ala Leu 275 280
285 Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr
Lys Thr Lys 290 295 300
Asn Val Val Glu Phe His Ser Asp Tyr Val Lys Val Lys Asn Ala Thr 305
310 315 320 Phe Leu Gly Val
Gln Met Lys Phe Ala Leu Gln Asn Leu Leu Lys Val 325
330 335 Ile Pro Asp Val Val Lys Gly Tyr Lys
Ser Val Pro Val Pro Thr Lys 340 345
350 Thr Pro Ala Asn Lys Gly Val Pro Ala Ser Thr Pro Leu Lys
Gln Glu 355 360 365
Trp Leu Trp Asn Glu Leu Ser Lys Phe Leu Gln Glu Gly Asp Val Ile 370
375 380 Ile Ser Glu Thr Gly
Thr Ser Ala Phe Gly Ile Asn Gln Thr Ile Phe 385 390
395 400 Pro Lys Asp Ala Tyr Gly Ile Ser Gln Val
Leu Trp Gly Ser Ile Gly 405 410
415 Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu
Ile 420 425 430 Asp
Pro Asn Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 435
440 445 Leu Thr Val Gln Glu Ile
Ser Thr Met Ile Arg Trp Gly Leu Lys Pro 450 455
460 Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr
Ile Glu Lys Leu Ile 465 470 475
480 His Gly Pro His Ala Glu Tyr Asn Glu Ile Gln Thr Trp Asp His Leu
485 490 495 Ala Leu
Leu Pro Ala Phe Gly Ala Lys Lys Tyr Glu Asn His Lys Ile 500
505 510 Ala Thr Thr Gly Glu Trp Asp
Ala Leu Thr Thr Asp Ser Glu Phe Gln 515 520
525 Lys Asn Ser Val Ile 530
1171695DNACandida glabrataCDS(1)..(1695) 117atg tct gag att act ttg ggt
aga tac ttg ttc gag aga ttg aac caa 48Met Ser Glu Ile Thr Leu Gly
Arg Tyr Leu Phe Glu Arg Leu Asn Gln 1 5
10 15 gtc gac gtt aag acc atc ttc
ggt ttg cca ggt gac ttc aac ttg tcc 96Val Asp Val Lys Thr Ile Phe
Gly Leu Pro Gly Asp Phe Asn Leu Ser 20
25 30 cta ttg gac aag atc tac gaa
gtt gaa ggt atg aga tgg gct ggt aac 144Leu Leu Asp Lys Ile Tyr Glu
Val Glu Gly Met Arg Trp Ala Gly Asn 35
40 45 gct aac gaa ttg aac gct gct
tac gct gct gac ggt tac gct aga atc 192Ala Asn Glu Leu Asn Ala Ala
Tyr Ala Ala Asp Gly Tyr Ala Arg Ile 50 55
60 aag ggt atg tcc tgt atc atc
acc acc ttc ggt gtc ggt gaa ttg tct 240Lys Gly Met Ser Cys Ile Ile
Thr Thr Phe Gly Val Gly Glu Leu Ser 65 70
75 80 gcc ttg aac ggt att gcc ggt
tct tac gct gaa cac gtc ggt gtc ttg 288Ala Leu Asn Gly Ile Ala Gly
Ser Tyr Ala Glu His Val Gly Val Leu 85
90 95 cac gtc gtc ggt gtc cca tcc
atc tcc tct caa gct aag caa ttg ttg 336His Val Val Gly Val Pro Ser
Ile Ser Ser Gln Ala Lys Gln Leu Leu 100
105 110 ttg cac cac acc ttg ggt aac
ggt gac ttc act gtc ttc cac aga atg 384Leu His His Thr Leu Gly Asn
Gly Asp Phe Thr Val Phe His Arg Met 115
120 125 tcc gct aac atc tct gag acc
acc gct atg gtc act gac atc gct acc 432Ser Ala Asn Ile Ser Glu Thr
Thr Ala Met Val Thr Asp Ile Ala Thr 130 135
140 gct cca gct gag atc gac aga
tgt atc aga acc acc tac atc acc caa 480Ala Pro Ala Glu Ile Asp Arg
Cys Ile Arg Thr Thr Tyr Ile Thr Gln 145 150
155 160 aga cca gtc tac ttg ggt cta
cca gct aac ttg gtc gac cta aag gtc 528Arg Pro Val Tyr Leu Gly Leu
Pro Ala Asn Leu Val Asp Leu Lys Val 165
170 175 cca gcc aag ctt ttg gaa acc
cca att gac ttg tcc ttg aag cca aac 576Pro Ala Lys Leu Leu Glu Thr
Pro Ile Asp Leu Ser Leu Lys Pro Asn 180
185 190 gac cca gaa gcc gaa act gaa
gtc gtt gac acc gtc ttg gaa ttg atc 624Asp Pro Glu Ala Glu Thr Glu
Val Val Asp Thr Val Leu Glu Leu Ile 195
200 205 aag gct gct aag aac cca gtt
atc ttg gct gat gct tgt gct tcc aga 672Lys Ala Ala Lys Asn Pro Val
Ile Leu Ala Asp Ala Cys Ala Ser Arg 210 215
220 cac gac gtc aag gct gaa acc
aag aag ttg att gac gcc act caa ttc 720His Asp Val Lys Ala Glu Thr
Lys Lys Leu Ile Asp Ala Thr Gln Phe 225 230
235 240 cca tcc ttc gtt acc cca atg
ggt aag ggt tcc atc gac gaa caa cac 768Pro Ser Phe Val Thr Pro Met
Gly Lys Gly Ser Ile Asp Glu Gln His 245
250 255 cca aga ttc ggt ggt gtc tac
gtc ggt acc ttg tcc aga cca gaa gtt 816Pro Arg Phe Gly Gly Val Tyr
Val Gly Thr Leu Ser Arg Pro Glu Val 260
265 270 aag gaa gct gtt gaa tcc gct
gac ttg atc ttg tct gtc ggt gct ttg 864Lys Glu Ala Val Glu Ser Ala
Asp Leu Ile Leu Ser Val Gly Ala Leu 275
280 285 ttg tcc gat ttc aac act ggt
tct ttc tct tac tct tac aag acc aag 912Leu Ser Asp Phe Asn Thr Gly
Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290 295
300 aac atc gtc gaa ttc cac tct
gac tac atc aag atc aga aac gct acc 960Asn Ile Val Glu Phe His Ser
Asp Tyr Ile Lys Ile Arg Asn Ala Thr 305 310
315 320 ttc cca ggt gtc caa atg aag
ttc gct ttg caa aag ttg ttg aac gcc 1008Phe Pro Gly Val Gln Met Lys
Phe Ala Leu Gln Lys Leu Leu Asn Ala 325
330 335 gtc cca gaa gct atc aag ggt
tac aag cca gtc cct gtc cca gct aga 1056Val Pro Glu Ala Ile Lys Gly
Tyr Lys Pro Val Pro Val Pro Ala Arg 340
345 350 gtc cca gaa aac aag tcc tgt
gac cca gct acc cca ttg aag caa gaa 1104Val Pro Glu Asn Lys Ser Cys
Asp Pro Ala Thr Pro Leu Lys Gln Glu 355
360 365 tgg atg tgg aac caa gtt tcc
aag ttc ttg caa gaa ggt gat gtt gtt 1152Trp Met Trp Asn Gln Val Ser
Lys Phe Leu Gln Glu Gly Asp Val Val 370 375
380 atc act gaa acc ggt acc tcc
gct ttt ggt atc aac caa acc cca ttc 1200Ile Thr Glu Thr Gly Thr Ser
Ala Phe Gly Ile Asn Gln Thr Pro Phe 385 390
395 400 cca aac aac gct tac ggt atc
tcc caa gtt cta tgg ggt tcc atc ggt 1248Pro Asn Asn Ala Tyr Gly Ile
Ser Gln Val Leu Trp Gly Ser Ile Gly 405
410 415 ttc acc acc ggt gct tgt ttg
ggt gcc gct ttc gct gct gaa gaa atc 1296Phe Thr Thr Gly Ala Cys Leu
Gly Ala Ala Phe Ala Ala Glu Glu Ile 420
425 430 gac cca aag aag aga gtt atc
ttg ttc att ggt gac ggt tct ttg caa 1344Asp Pro Lys Lys Arg Val Ile
Leu Phe Ile Gly Asp Gly Ser Leu Gln 435
440 445 ttg act gtc caa gaa atc tcc
acc atg atc aga tgg ggc ttg aag cca 1392Leu Thr Val Gln Glu Ile Ser
Thr Met Ile Arg Trp Gly Leu Lys Pro 450 455
460 tac ttg ttc gtc ttg aac aac
gac ggt tac acc atc gaa aga ttg att 1440Tyr Leu Phe Val Leu Asn Asn
Asp Gly Tyr Thr Ile Glu Arg Leu Ile 465 470
475 480 cac ggt gaa aag gct ggt tac
aac gac atc caa aac tgg gac cac ttg 1488His Gly Glu Lys Ala Gly Tyr
Asn Asp Ile Gln Asn Trp Asp His Leu 485
490 495 gct cta ttg cca acc ttc ggt
gct aag gac tac gaa aac cac aga gtc 1536Ala Leu Leu Pro Thr Phe Gly
Ala Lys Asp Tyr Glu Asn His Arg Val 500
505 510 gcc acc acc ggt gaa tgg gac
aag ttg acc caa gac aag gaa ttc aac 1584Ala Thr Thr Gly Glu Trp Asp
Lys Leu Thr Gln Asp Lys Glu Phe Asn 515
520 525 aag aac tcc aag atc aga atg
atc gaa gtt atg ttg cca gtt atg gac 1632Lys Asn Ser Lys Ile Arg Met
Ile Glu Val Met Leu Pro Val Met Asp 530 535
540 gct cca act tcc ttg att gaa
caa gct aag ttg acc gct tcc atc aac 1680Ala Pro Thr Ser Leu Ile Glu
Gln Ala Lys Leu Thr Ala Ser Ile Asn 545 550
555 560 gct aag caa gaa taa
1695Ala Lys Gln Glu
118564PRTCandida glabrata
118Met Ser Glu Ile Thr Leu Gly Arg Tyr Leu Phe Glu Arg Leu Asn Gln 1
5 10 15 Val Asp Val Lys
Thr Ile Phe Gly Leu Pro Gly Asp Phe Asn Leu Ser 20
25 30 Leu Leu Asp Lys Ile Tyr Glu Val Glu
Gly Met Arg Trp Ala Gly Asn 35 40
45 Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala
Arg Ile 50 55 60
Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser 65
70 75 80 Ala Leu Asn Gly Ile
Ala Gly Ser Tyr Ala Glu His Val Gly Val Leu 85
90 95 His Val Val Gly Val Pro Ser Ile Ser Ser
Gln Ala Lys Gln Leu Leu 100 105
110 Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg
Met 115 120 125 Ser
Ala Asn Ile Ser Glu Thr Thr Ala Met Val Thr Asp Ile Ala Thr 130
135 140 Ala Pro Ala Glu Ile Asp
Arg Cys Ile Arg Thr Thr Tyr Ile Thr Gln 145 150
155 160 Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu
Val Asp Leu Lys Val 165 170
175 Pro Ala Lys Leu Leu Glu Thr Pro Ile Asp Leu Ser Leu Lys Pro Asn
180 185 190 Asp Pro
Glu Ala Glu Thr Glu Val Val Asp Thr Val Leu Glu Leu Ile 195
200 205 Lys Ala Ala Lys Asn Pro Val
Ile Leu Ala Asp Ala Cys Ala Ser Arg 210 215
220 His Asp Val Lys Ala Glu Thr Lys Lys Leu Ile Asp
Ala Thr Gln Phe 225 230 235
240 Pro Ser Phe Val Thr Pro Met Gly Lys Gly Ser Ile Asp Glu Gln His
245 250 255 Pro Arg Phe
Gly Gly Val Tyr Val Gly Thr Leu Ser Arg Pro Glu Val 260
265 270 Lys Glu Ala Val Glu Ser Ala Asp
Leu Ile Leu Ser Val Gly Ala Leu 275 280
285 Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr
Lys Thr Lys 290 295 300
Asn Ile Val Glu Phe His Ser Asp Tyr Ile Lys Ile Arg Asn Ala Thr 305
310 315 320 Phe Pro Gly Val
Gln Met Lys Phe Ala Leu Gln Lys Leu Leu Asn Ala 325
330 335 Val Pro Glu Ala Ile Lys Gly Tyr Lys
Pro Val Pro Val Pro Ala Arg 340 345
350 Val Pro Glu Asn Lys Ser Cys Asp Pro Ala Thr Pro Leu Lys
Gln Glu 355 360 365
Trp Met Trp Asn Gln Val Ser Lys Phe Leu Gln Glu Gly Asp Val Val 370
375 380 Ile Thr Glu Thr Gly
Thr Ser Ala Phe Gly Ile Asn Gln Thr Pro Phe 385 390
395 400 Pro Asn Asn Ala Tyr Gly Ile Ser Gln Val
Leu Trp Gly Ser Ile Gly 405 410
415 Phe Thr Thr Gly Ala Cys Leu Gly Ala Ala Phe Ala Ala Glu Glu
Ile 420 425 430 Asp
Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 435
440 445 Leu Thr Val Gln Glu Ile
Ser Thr Met Ile Arg Trp Gly Leu Lys Pro 450 455
460 Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr
Ile Glu Arg Leu Ile 465 470 475
480 His Gly Glu Lys Ala Gly Tyr Asn Asp Ile Gln Asn Trp Asp His Leu
485 490 495 Ala Leu
Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu Asn His Arg Val 500
505 510 Ala Thr Thr Gly Glu Trp Asp
Lys Leu Thr Gln Asp Lys Glu Phe Asn 515 520
525 Lys Asn Ser Lys Ile Arg Met Ile Glu Val Met Leu
Pro Val Met Asp 530 535 540
Ala Pro Thr Ser Leu Ile Glu Gln Ala Lys Leu Thr Ala Ser Ile Asn 545
550 555 560 Ala Lys Gln
Glu 1191791DNAPichia stipitisCDS(1)..(1791) 119atg gct gaa gtc tca tta
gga aga tat ctc ttc gag aga ttg tac caa 48Met Ala Glu Val Ser Leu
Gly Arg Tyr Leu Phe Glu Arg Leu Tyr Gln 1 5
10 15 ttg caa gtg cag acc atc
ttc ggt gtc cct ggt gat ttc aac ttg tcg 96Leu Gln Val Gln Thr Ile
Phe Gly Val Pro Gly Asp Phe Asn Leu Ser 20
25 30 ctt ttg gac aag atc tac
gaa gtg gaa gat gcc cat ggc aag aat tcg 144Leu Leu Asp Lys Ile Tyr
Glu Val Glu Asp Ala His Gly Lys Asn Ser 35
40 45 ttt aga tgg gct ggt aat
gcc aac gaa ttg aat gca tcg tac gct gct 192Phe Arg Trp Ala Gly Asn
Ala Asn Glu Leu Asn Ala Ser Tyr Ala Ala 50
55 60 gac ggt tac tcg aga gtc
aag cgt tta ggg tgt ttg gtc act acc ttt 240Asp Gly Tyr Ser Arg Val
Lys Arg Leu Gly Cys Leu Val Thr Thr Phe 65 70
75 80 ggt gtc ggt gaa ttg tct
gct ttg aat ggt att gcc ggt tct tat gcc 288Gly Val Gly Glu Leu Ser
Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala 85
90 95 gaa cat gtt ggt ttg ctt
cat gtc gta ggt gtt cca tcg att tcc tcg 336Glu His Val Gly Leu Leu
His Val Val Gly Val Pro Ser Ile Ser Ser 100
105 110 caa gct aag caa ttg tta
ctt cac cac act ttg ggt aat ggt gat ttc 384Gln Ala Lys Gln Leu Leu
Leu His His Thr Leu Gly Asn Gly Asp Phe 115
120 125 act gtt ttc cat aga atg
tcc aac aac att tct cag acc aca gcc ttt 432Thr Val Phe His Arg Met
Ser Asn Asn Ile Ser Gln Thr Thr Ala Phe 130
135 140 atc tcc gat atc aac tcg
gct cca gct gaa att gat aga tgt atc aga 480Ile Ser Asp Ile Asn Ser
Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg 145 150
155 160 gag gcc tac gtc aaa caa
aga cca gtt tat atc ggg tta cca gct aac 528Glu Ala Tyr Val Lys Gln
Arg Pro Val Tyr Ile Gly Leu Pro Ala Asn 165
170 175 tta gtt gat ttg aat gtt
ccg gcc tct ttg ctt gag tct cca atc aac 576Leu Val Asp Leu Asn Val
Pro Ala Ser Leu Leu Glu Ser Pro Ile Asn 180
185 190 ttg tcg ttg gaa aag aac
gac cca gag gct caa gat gaa gtc att gac 624Leu Ser Leu Glu Lys Asn
Asp Pro Glu Ala Gln Asp Glu Val Ile Asp 195
200 205 tct gtc tta gac ttg atc
aaa aag tcg ctg aac cca atc atc ttg gtc 672Ser Val Leu Asp Leu Ile
Lys Lys Ser Leu Asn Pro Ile Ile Leu Val 210
215 220 gat gcc tgt gcc tcg aga
cat gac tgt aag gct gaa gtt act cag ttg 720Asp Ala Cys Ala Ser Arg
His Asp Cys Lys Ala Glu Val Thr Gln Leu 225 230
235 240 att gaa caa acc caa ttc
cca gta ttt gtc act cca atg ggt aaa ggt 768Ile Glu Gln Thr Gln Phe
Pro Val Phe Val Thr Pro Met Gly Lys Gly 245
250 255 acc gtt gat gag ggt ggt
gta gac gga gaa ttg tta gaa gat gat cct 816Thr Val Asp Glu Gly Gly
Val Asp Gly Glu Leu Leu Glu Asp Asp Pro 260
265 270 cat ttg att gcc aag gtc
gct gct agg ttg tct gct ggc aag aac gct 864His Leu Ile Ala Lys Val
Ala Ala Arg Leu Ser Ala Gly Lys Asn Ala 275
280 285 gcc tct aga ttc gga ggt
gtt tat gtc gga acc ttg tcg aag ccc gaa 912Ala Ser Arg Phe Gly Gly
Val Tyr Val Gly Thr Leu Ser Lys Pro Glu 290
295 300 gtc aag gac gct gta gag
agt gca gat ttg att ttg tct gtc ggt gcc 960Val Lys Asp Ala Val Glu
Ser Ala Asp Leu Ile Leu Ser Val Gly Ala 305 310
315 320 ctt ttg tct gat ttc aac
act ggt tca ttt tcc tac tcc tac aga acc 1008Leu Leu Ser Asp Phe Asn
Thr Gly Ser Phe Ser Tyr Ser Tyr Arg Thr 325
330 335 aag aac atc gtc gaa ttc
cat tct gat tac act aag att aga caa gcc 1056Lys Asn Ile Val Glu Phe
His Ser Asp Tyr Thr Lys Ile Arg Gln Ala 340
345 350 act ttc cca ggt gtg cag
atg aag gaa gcc ttg caa gaa ttg aac aag 1104Thr Phe Pro Gly Val Gln
Met Lys Glu Ala Leu Gln Glu Leu Asn Lys 355
360 365 aaa gtt tca tct gct gct
agt cac tat gaa gtc aag cct gtg ccc aag 1152Lys Val Ser Ser Ala Ala
Ser His Tyr Glu Val Lys Pro Val Pro Lys 370
375 380 atc aag ttg gcc aat aca
cca gcc acc aga gaa gtc aag tta act cag 1200Ile Lys Leu Ala Asn Thr
Pro Ala Thr Arg Glu Val Lys Leu Thr Gln 385 390
395 400 gaa tgg ttg tgg acc aga
gtg tct tcg tgg ttc aga gaa ggt gat att 1248Glu Trp Leu Trp Thr Arg
Val Ser Ser Trp Phe Arg Glu Gly Asp Ile 405
410 415 att atc acc gaa acc ggt
aca tcc tcc ttc ggt ata gtt caa tcc aga 1296Ile Ile Thr Glu Thr Gly
Thr Ser Ser Phe Gly Ile Val Gln Ser Arg 420
425 430 ttc cca aac aac acc atc
ggt atc tcc caa gta ttg tgg ggt tct att 1344Phe Pro Asn Asn Thr Ile
Gly Ile Ser Gln Val Leu Trp Gly Ser Ile 435
440 445 ggt ttc tct gtt ggt gcc
act ttg ggt gct gcc atg gct gcc caa gaa 1392Gly Phe Ser Val Gly Ala
Thr Leu Gly Ala Ala Met Ala Ala Gln Glu 450
455 460 ctc gac cct aac aag aga
acc atc ttg ttt gtt gga gat ggt tct ttg 1440Leu Asp Pro Asn Lys Arg
Thr Ile Leu Phe Val Gly Asp Gly Ser Leu 465 470
475 480 caa ttg acc gtt cag gaa
atc tcc acc ata atc aga tgg ggt acc aca 1488Gln Leu Thr Val Gln Glu
Ile Ser Thr Ile Ile Arg Trp Gly Thr Thr 485
490 495 cct tac ctt ttc gtg ttg
aac aat gac ggt tac acc atc gag cgt ttg 1536Pro Tyr Leu Phe Val Leu
Asn Asn Asp Gly Tyr Thr Ile Glu Arg Leu 500
505 510 atc cac ggt gta aat gcc
tca tat aat gac atc caa cca tgg caa aac 1584Ile His Gly Val Asn Ala
Ser Tyr Asn Asp Ile Gln Pro Trp Gln Asn 515
520 525 ttg gaa atc ttg cct act
ttc tcg gcc aag aac tac gac gct gtg aga 1632Leu Glu Ile Leu Pro Thr
Phe Ser Ala Lys Asn Tyr Asp Ala Val Arg 530
535 540 atc tcc aac atc gga gaa
gca gaa gat atc ttg aaa gac aag gaa ttc 1680Ile Ser Asn Ile Gly Glu
Ala Glu Asp Ile Leu Lys Asp Lys Glu Phe 545 550
555 560 gga aag aac tcc aag att
aga ttg ata gaa gtc atg tta cca aga ttg 1728Gly Lys Asn Ser Lys Ile
Arg Leu Ile Glu Val Met Leu Pro Arg Leu 565
570 575 gat gca cca tct aac ctt
gcc aaa caa gct gcc att aca gct gcc acc 1776Asp Ala Pro Ser Asn Leu
Ala Lys Gln Ala Ala Ile Thr Ala Ala Thr 580
585 590 aac gcc gaa gct tag
1791Asn Ala Glu Ala
595
120596PRTPichia stipitis
120Met Ala Glu Val Ser Leu Gly Arg Tyr Leu Phe Glu Arg Leu Tyr Gln 1
5 10 15 Leu Gln Val Gln
Thr Ile Phe Gly Val Pro Gly Asp Phe Asn Leu Ser 20
25 30 Leu Leu Asp Lys Ile Tyr Glu Val Glu
Asp Ala His Gly Lys Asn Ser 35 40
45 Phe Arg Trp Ala Gly Asn Ala Asn Glu Leu Asn Ala Ser Tyr
Ala Ala 50 55 60
Asp Gly Tyr Ser Arg Val Lys Arg Leu Gly Cys Leu Val Thr Thr Phe 65
70 75 80 Gly Val Gly Glu Leu
Ser Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala 85
90 95 Glu His Val Gly Leu Leu His Val Val Gly
Val Pro Ser Ile Ser Ser 100 105
110 Gln Ala Lys Gln Leu Leu Leu His His Thr Leu Gly Asn Gly Asp
Phe 115 120 125 Thr
Val Phe His Arg Met Ser Asn Asn Ile Ser Gln Thr Thr Ala Phe 130
135 140 Ile Ser Asp Ile Asn Ser
Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg 145 150
155 160 Glu Ala Tyr Val Lys Gln Arg Pro Val Tyr Ile
Gly Leu Pro Ala Asn 165 170
175 Leu Val Asp Leu Asn Val Pro Ala Ser Leu Leu Glu Ser Pro Ile Asn
180 185 190 Leu Ser
Leu Glu Lys Asn Asp Pro Glu Ala Gln Asp Glu Val Ile Asp 195
200 205 Ser Val Leu Asp Leu Ile Lys
Lys Ser Leu Asn Pro Ile Ile Leu Val 210 215
220 Asp Ala Cys Ala Ser Arg His Asp Cys Lys Ala Glu
Val Thr Gln Leu 225 230 235
240 Ile Glu Gln Thr Gln Phe Pro Val Phe Val Thr Pro Met Gly Lys Gly
245 250 255 Thr Val Asp
Glu Gly Gly Val Asp Gly Glu Leu Leu Glu Asp Asp Pro 260
265 270 His Leu Ile Ala Lys Val Ala Ala
Arg Leu Ser Ala Gly Lys Asn Ala 275 280
285 Ala Ser Arg Phe Gly Gly Val Tyr Val Gly Thr Leu Ser
Lys Pro Glu 290 295 300
Val Lys Asp Ala Val Glu Ser Ala Asp Leu Ile Leu Ser Val Gly Ala 305
310 315 320 Leu Leu Ser Asp
Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Arg Thr 325
330 335 Lys Asn Ile Val Glu Phe His Ser Asp
Tyr Thr Lys Ile Arg Gln Ala 340 345
350 Thr Phe Pro Gly Val Gln Met Lys Glu Ala Leu Gln Glu Leu
Asn Lys 355 360 365
Lys Val Ser Ser Ala Ala Ser His Tyr Glu Val Lys Pro Val Pro Lys 370
375 380 Ile Lys Leu Ala Asn
Thr Pro Ala Thr Arg Glu Val Lys Leu Thr Gln 385 390
395 400 Glu Trp Leu Trp Thr Arg Val Ser Ser Trp
Phe Arg Glu Gly Asp Ile 405 410
415 Ile Ile Thr Glu Thr Gly Thr Ser Ser Phe Gly Ile Val Gln Ser
Arg 420 425 430 Phe
Pro Asn Asn Thr Ile Gly Ile Ser Gln Val Leu Trp Gly Ser Ile 435
440 445 Gly Phe Ser Val Gly Ala
Thr Leu Gly Ala Ala Met Ala Ala Gln Glu 450 455
460 Leu Asp Pro Asn Lys Arg Thr Ile Leu Phe Val
Gly Asp Gly Ser Leu 465 470 475
480 Gln Leu Thr Val Gln Glu Ile Ser Thr Ile Ile Arg Trp Gly Thr Thr
485 490 495 Pro Tyr
Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile Glu Arg Leu 500
505 510 Ile His Gly Val Asn Ala Ser
Tyr Asn Asp Ile Gln Pro Trp Gln Asn 515 520
525 Leu Glu Ile Leu Pro Thr Phe Ser Ala Lys Asn Tyr
Asp Ala Val Arg 530 535 540
Ile Ser Asn Ile Gly Glu Ala Glu Asp Ile Leu Lys Asp Lys Glu Phe 545
550 555 560 Gly Lys Asn
Ser Lys Ile Arg Leu Ile Glu Val Met Leu Pro Arg Leu 565
570 575 Asp Ala Pro Ser Asn Leu Ala Lys
Gln Ala Ala Ile Thr Ala Ala Thr 580 585
590 Asn Ala Glu Ala 595 1211710DNAPichia
stipitisCDS(1)..(1710) 121atg gta tca acc tac cca gaa tca gag gtt act cta
gga agg tac ctc 48Met Val Ser Thr Tyr Pro Glu Ser Glu Val Thr Leu
Gly Arg Tyr Leu 1 5 10
15 ttt gag cga ctc cac caa ttg aaa gtg gac acc att
ttc ggc ttg ccg 96Phe Glu Arg Leu His Gln Leu Lys Val Asp Thr Ile
Phe Gly Leu Pro 20 25
30 ggt gac ttc aac ctt tcc tta ttg gac aaa gtg tat
gaa gtt ccg gat 144Gly Asp Phe Asn Leu Ser Leu Leu Asp Lys Val Tyr
Glu Val Pro Asp 35 40
45 atg agg tgg gct gga aat gcc aac gaa ttg aat gct
gcc tat gct gcc 192Met Arg Trp Ala Gly Asn Ala Asn Glu Leu Asn Ala
Ala Tyr Ala Ala 50 55 60
gat ggt tac tcc aga ata aag gga ttg tct tgc ttg
gtc aca act ttt 240Asp Gly Tyr Ser Arg Ile Lys Gly Leu Ser Cys Leu
Val Thr Thr Phe 65 70 75
80 ggt gtt ggt gaa ttg tct gct tta aac gga gtt ggt
ggt gcc tat gct 288Gly Val Gly Glu Leu Ser Ala Leu Asn Gly Val Gly
Gly Ala Tyr Ala 85 90
95 gaa cac gta gga ctt cta cat gtc gtt gga gtt cca
tcc ata tcg tca 336Glu His Val Gly Leu Leu His Val Val Gly Val Pro
Ser Ile Ser Ser 100 105
110 cag gct aaa cag ttg ttg ctc cac cat acc ttg ggt
aat ggt gac ttc 384Gln Ala Lys Gln Leu Leu Leu His His Thr Leu Gly
Asn Gly Asp Phe 115 120
125 act gtt ttt cac aga atg tcc aat agc att tct caa
act aca gca ttt 432Thr Val Phe His Arg Met Ser Asn Ser Ile Ser Gln
Thr Thr Ala Phe 130 135 140
ctc tca gat atc tct att gca cca ggt caa ata gat
aga tgc atc aga 480Leu Ser Asp Ile Ser Ile Ala Pro Gly Gln Ile Asp
Arg Cys Ile Arg 145 150 155
160 gaa gca tat gtt cat cag aga cca gtt tat gtt ggt
tta ccg gca aat 528Glu Ala Tyr Val His Gln Arg Pro Val Tyr Val Gly
Leu Pro Ala Asn 165 170
175 atg gtt gat ctc aag gtt cct tct agt ctc tta gaa
act cca att gat 576Met Val Asp Leu Lys Val Pro Ser Ser Leu Leu Glu
Thr Pro Ile Asp 180 185
190 ttg aaa ttg aaa caa aat gat cct gaa gct caa gaa
gtt gtt gaa aca 624Leu Lys Leu Lys Gln Asn Asp Pro Glu Ala Gln Glu
Val Val Glu Thr 195 200
205 gtc ctg aag ttg gtg tcc caa gct aca aac ccc att
atc ttg gta gac 672Val Leu Lys Leu Val Ser Gln Ala Thr Asn Pro Ile
Ile Leu Val Asp 210 215 220
gct tgt gcc ctc aga cac aat tgc aaa gag gaa gtc
aaa caa ttg gtt 720Ala Cys Ala Leu Arg His Asn Cys Lys Glu Glu Val
Lys Gln Leu Val 225 230 235
240 gat gcc act aat ttt caa gtc ttt aca act cca atg
ggt aaa tct ggt 768Asp Ala Thr Asn Phe Gln Val Phe Thr Thr Pro Met
Gly Lys Ser Gly 245 250
255 atc tcc gaa tct cat cca aga ttg ggc ggt gtc tat
gtc ggg aca atg 816Ile Ser Glu Ser His Pro Arg Leu Gly Gly Val Tyr
Val Gly Thr Met 260 265
270 tcg agt cct caa gtc aaa aaa gcc gtt gaa aat gcc
gat ctt ata cta 864Ser Ser Pro Gln Val Lys Lys Ala Val Glu Asn Ala
Asp Leu Ile Leu 275 280
285 tct gtt ggt tcg ttg tta tcg gac ttc aat aca ggt
tca ttt tca tac 912Ser Val Gly Ser Leu Leu Ser Asp Phe Asn Thr Gly
Ser Phe Ser Tyr 290 295 300
tcc tac aag acg aag aat gtt gtt gaa ttc cac tct
gac tat atg aaa 960Ser Tyr Lys Thr Lys Asn Val Val Glu Phe His Ser
Asp Tyr Met Lys 305 310 315
320 atc aga cag gcc acc ttc cca gga gtt caa atg aaa
gaa gcc ttg caa 1008Ile Arg Gln Ala Thr Phe Pro Gly Val Gln Met Lys
Glu Ala Leu Gln 325 330
335 cag ttg ata aaa agg gtc tct tct tac atc aat cca
agc tac att cct 1056Gln Leu Ile Lys Arg Val Ser Ser Tyr Ile Asn Pro
Ser Tyr Ile Pro 340 345
350 act cga gtt cct aaa agg aaa cag cca ttg aaa gct
cca tca gaa gct 1104Thr Arg Val Pro Lys Arg Lys Gln Pro Leu Lys Ala
Pro Ser Glu Ala 355 360
365 cct ttg acc caa gaa tat ttg tgg tct aaa gta tcc
ggc tgg ttt aga 1152Pro Leu Thr Gln Glu Tyr Leu Trp Ser Lys Val Ser
Gly Trp Phe Arg 370 375 380
gag ggt gat att atc gta acc gaa act ggt aca tct
gct ttc gga att 1200Glu Gly Asp Ile Ile Val Thr Glu Thr Gly Thr Ser
Ala Phe Gly Ile 385 390 395
400 att caa tcc cat ttt ccc agc aac act atc ggt ata
tcc caa gtc ttg 1248Ile Gln Ser His Phe Pro Ser Asn Thr Ile Gly Ile
Ser Gln Val Leu 405 410
415 tgg ggc tca att ggt ttc aca gta ggt gca aca gtt
ggt gct gcc atg 1296Trp Gly Ser Ile Gly Phe Thr Val Gly Ala Thr Val
Gly Ala Ala Met 420 425
430 gca gcc cag gaa atc gac cct agc agg aga gta att
ttg ttc gtc ggt 1344Ala Ala Gln Glu Ile Asp Pro Ser Arg Arg Val Ile
Leu Phe Val Gly 435 440
445 gat ggt tca ttg cag ttg acg gtt cag gaa atc tct
acg ttg tgt aaa 1392Asp Gly Ser Leu Gln Leu Thr Val Gln Glu Ile Ser
Thr Leu Cys Lys 450 455 460
tgg gat tgt aac aat act tat ctt tac gtg ttg aac
aat gat ggt tac 1440Trp Asp Cys Asn Asn Thr Tyr Leu Tyr Val Leu Asn
Asn Asp Gly Tyr 465 470 475
480 act ata gaa agg ttg atc cac ggc aaa agt gcc agc
tac aac gat ata 1488Thr Ile Glu Arg Leu Ile His Gly Lys Ser Ala Ser
Tyr Asn Asp Ile 485 490
495 cag cct tgg aac cat tta tcc ttg ctt cgc tta ttc
aat gct aag aaa 1536Gln Pro Trp Asn His Leu Ser Leu Leu Arg Leu Phe
Asn Ala Lys Lys 500 505
510 tac caa aat gtc aga gta tcg act gct gga gaa ttg
gac tct ttg ttc 1584Tyr Gln Asn Val Arg Val Ser Thr Ala Gly Glu Leu
Asp Ser Leu Phe 515 520
525 tct gat aag aaa ttt gct tct cca gat agg ata aga
atg att gag gtg 1632Ser Asp Lys Lys Phe Ala Ser Pro Asp Arg Ile Arg
Met Ile Glu Val 530 535 540
atg tta tcg aga ttg gat gca cca gca aat ctt gtt
gct caa gca aag 1680Met Leu Ser Arg Leu Asp Ala Pro Ala Asn Leu Val
Ala Gln Ala Lys 545 550 555
560 ttg tct gaa cgg gta aac ctt gaa aat tga
1710Leu Ser Glu Arg Val Asn Leu Glu Asn
565
122569PRTPichia stipitis 122Met Val Ser Thr Tyr
Pro Glu Ser Glu Val Thr Leu Gly Arg Tyr Leu 1 5
10 15 Phe Glu Arg Leu His Gln Leu Lys Val Asp
Thr Ile Phe Gly Leu Pro 20 25
30 Gly Asp Phe Asn Leu Ser Leu Leu Asp Lys Val Tyr Glu Val Pro
Asp 35 40 45 Met
Arg Trp Ala Gly Asn Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala 50
55 60 Asp Gly Tyr Ser Arg Ile
Lys Gly Leu Ser Cys Leu Val Thr Thr Phe 65 70
75 80 Gly Val Gly Glu Leu Ser Ala Leu Asn Gly Val
Gly Gly Ala Tyr Ala 85 90
95 Glu His Val Gly Leu Leu His Val Val Gly Val Pro Ser Ile Ser Ser
100 105 110 Gln Ala
Lys Gln Leu Leu Leu His His Thr Leu Gly Asn Gly Asp Phe 115
120 125 Thr Val Phe His Arg Met Ser
Asn Ser Ile Ser Gln Thr Thr Ala Phe 130 135
140 Leu Ser Asp Ile Ser Ile Ala Pro Gly Gln Ile Asp
Arg Cys Ile Arg 145 150 155
160 Glu Ala Tyr Val His Gln Arg Pro Val Tyr Val Gly Leu Pro Ala Asn
165 170 175 Met Val Asp
Leu Lys Val Pro Ser Ser Leu Leu Glu Thr Pro Ile Asp 180
185 190 Leu Lys Leu Lys Gln Asn Asp Pro
Glu Ala Gln Glu Val Val Glu Thr 195 200
205 Val Leu Lys Leu Val Ser Gln Ala Thr Asn Pro Ile Ile
Leu Val Asp 210 215 220
Ala Cys Ala Leu Arg His Asn Cys Lys Glu Glu Val Lys Gln Leu Val 225
230 235 240 Asp Ala Thr Asn
Phe Gln Val Phe Thr Thr Pro Met Gly Lys Ser Gly 245
250 255 Ile Ser Glu Ser His Pro Arg Leu Gly
Gly Val Tyr Val Gly Thr Met 260 265
270 Ser Ser Pro Gln Val Lys Lys Ala Val Glu Asn Ala Asp Leu
Ile Leu 275 280 285
Ser Val Gly Ser Leu Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr 290
295 300 Ser Tyr Lys Thr Lys
Asn Val Val Glu Phe His Ser Asp Tyr Met Lys 305 310
315 320 Ile Arg Gln Ala Thr Phe Pro Gly Val Gln
Met Lys Glu Ala Leu Gln 325 330
335 Gln Leu Ile Lys Arg Val Ser Ser Tyr Ile Asn Pro Ser Tyr Ile
Pro 340 345 350 Thr
Arg Val Pro Lys Arg Lys Gln Pro Leu Lys Ala Pro Ser Glu Ala 355
360 365 Pro Leu Thr Gln Glu Tyr
Leu Trp Ser Lys Val Ser Gly Trp Phe Arg 370 375
380 Glu Gly Asp Ile Ile Val Thr Glu Thr Gly Thr
Ser Ala Phe Gly Ile 385 390 395
400 Ile Gln Ser His Phe Pro Ser Asn Thr Ile Gly Ile Ser Gln Val Leu
405 410 415 Trp Gly
Ser Ile Gly Phe Thr Val Gly Ala Thr Val Gly Ala Ala Met 420
425 430 Ala Ala Gln Glu Ile Asp Pro
Ser Arg Arg Val Ile Leu Phe Val Gly 435 440
445 Asp Gly Ser Leu Gln Leu Thr Val Gln Glu Ile Ser
Thr Leu Cys Lys 450 455 460
Trp Asp Cys Asn Asn Thr Tyr Leu Tyr Val Leu Asn Asn Asp Gly Tyr 465
470 475 480 Thr Ile Glu
Arg Leu Ile His Gly Lys Ser Ala Ser Tyr Asn Asp Ile 485
490 495 Gln Pro Trp Asn His Leu Ser Leu
Leu Arg Leu Phe Asn Ala Lys Lys 500 505
510 Tyr Gln Asn Val Arg Val Ser Thr Ala Gly Glu Leu Asp
Ser Leu Phe 515 520 525
Ser Asp Lys Lys Phe Ala Ser Pro Asp Arg Ile Arg Met Ile Glu Val 530
535 540 Met Leu Ser Arg
Leu Asp Ala Pro Ala Asn Leu Val Ala Gln Ala Lys 545 550
555 560 Leu Ser Glu Arg Val Asn Leu Glu Asn
565 1231692DNAKluyveromyces
lactisCDS(1)..(1692) 123atg tct gaa att aca tta ggt cgt tac ttg ttc gaa
aga tta aag caa 48Met Ser Glu Ile Thr Leu Gly Arg Tyr Leu Phe Glu
Arg Leu Lys Gln 1 5 10
15 gtc gaa gtt caa acc atc ttt ggt cta cca ggt gat
ttc aac ttg tcc 96Val Glu Val Gln Thr Ile Phe Gly Leu Pro Gly Asp
Phe Asn Leu Ser 20 25
30 cta ttg gac aat atc tac gaa gtc cca ggt atg aga
tgg gct ggt aat 144Leu Leu Asp Asn Ile Tyr Glu Val Pro Gly Met Arg
Trp Ala Gly Asn 35 40
45 gcc aac gaa ttg aac gct gct tac gct gct gat ggt
tac gcc aga tta 192Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly
Tyr Ala Arg Leu 50 55 60
aag ggt atg tcc tgt atc atc acc acc ttc ggt gtc
ggt gaa ttg tct 240Lys Gly Met Ser Cys Ile Ile Thr Thr Phe Gly Val
Gly Glu Leu Ser 65 70 75
80 gct ttg aac ggt att gcc ggt tct tac gct gaa cac
gtt ggt gtc ttg 288Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala Glu His
Val Gly Val Leu 85 90
95 cac gtt gtc ggt gtt cca tcc gtc tct tct caa gct
aag caa ttg ttg 336His Val Val Gly Val Pro Ser Val Ser Ser Gln Ala
Lys Gln Leu Leu 100 105
110 ttg cac cac acc ttg ggt aac ggt gac ttc act gtt
ttc cac aga atg 384Leu His His Thr Leu Gly Asn Gly Asp Phe Thr Val
Phe His Arg Met 115 120
125 tcc tcc aac att tct gaa acc act gct atg atc acc
gat atc aac act 432Ser Ser Asn Ile Ser Glu Thr Thr Ala Met Ile Thr
Asp Ile Asn Thr 130 135 140
gcc cca gct gaa atc gac aga tgt atc aga acc act
tac gtt tcc caa 480Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr Thr
Tyr Val Ser Gln 145 150 155
160 aga cca gtc tac ttg ggt ttg cca gct aac ttg gtc
gac ttg act gtc 528Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val
Asp Leu Thr Val 165 170
175 cca gct tct ttg ttg gac act cca att gat ttg agc
ttg aag cca aat 576Pro Ala Ser Leu Leu Asp Thr Pro Ile Asp Leu Ser
Leu Lys Pro Asn 180 185
190 gac cca gaa gcc gaa gaa gaa gtc atc gaa aac gtc
ttg caa ctg atc 624Asp Pro Glu Ala Glu Glu Glu Val Ile Glu Asn Val
Leu Gln Leu Ile 195 200
205 aag gaa gct aag aac cca gtt atc ttg gct gat gct
tgt tgt tcc aga 672Lys Glu Ala Lys Asn Pro Val Ile Leu Ala Asp Ala
Cys Cys Ser Arg 210 215 220
cac gat gcc aag gct gag acc aag aag ttg atc gac
ttg act caa ttc 720His Asp Ala Lys Ala Glu Thr Lys Lys Leu Ile Asp
Leu Thr Gln Phe 225 230 235
240 cca gcc ttc gtt acc cca atg ggt aag ggt tcc att
gac gaa aag cac 768Pro Ala Phe Val Thr Pro Met Gly Lys Gly Ser Ile
Asp Glu Lys His 245 250
255 cca aga ttc ggt ggt gtc tac gtc ggt acc cta tct
tct cca gct gtc 816Pro Arg Phe Gly Gly Val Tyr Val Gly Thr Leu Ser
Ser Pro Ala Val 260 265
270 aag gaa gcc gtt gaa tct gct gac ttg gtt cta tcg
gtc ggt gct cta 864Lys Glu Ala Val Glu Ser Ala Asp Leu Val Leu Ser
Val Gly Ala Leu 275 280
285 ttg tcc gat ttc aac act ggt tct ttc tct tac tct
tac aag acc aag 912Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser
Tyr Lys Thr Lys 290 295 300
aac att gtc gaa ttc cac tct gac tac acc aag atc
aga agc gct acc 960Asn Ile Val Glu Phe His Ser Asp Tyr Thr Lys Ile
Arg Ser Ala Thr 305 310 315
320 ttc cca ggt gtc caa atg aag ttc gct tta caa aaa
ttg ttg act aag 1008Phe Pro Gly Val Gln Met Lys Phe Ala Leu Gln Lys
Leu Leu Thr Lys 325 330
335 gtt gcc gat gct gct aag ggt tac aag cca gtt cca
gtt cca tct gaa 1056Val Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Pro
Val Pro Ser Glu 340 345
350 cca gaa cac aac gaa gct gtc gct gac tcc act cca
ttg aag caa gaa 1104Pro Glu His Asn Glu Ala Val Ala Asp Ser Thr Pro
Leu Lys Gln Glu 355 360
365 tgg gtc tgg act caa gtc ggt gaa ttc ttg aga gaa
ggt gat gtt gtt 1152Trp Val Trp Thr Gln Val Gly Glu Phe Leu Arg Glu
Gly Asp Val Val 370 375 380
atc act gaa acc ggt acc tct gcc ttc ggt atc aac
caa act cat ttc 1200Ile Thr Glu Thr Gly Thr Ser Ala Phe Gly Ile Asn
Gln Thr His Phe 385 390 395
400 cca aac aac aca tac ggt atc tct caa gtt tta tgg
ggt tcc att ggt 1248Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val Leu Trp
Gly Ser Ile Gly 405 410
415 ttc acc act ggt gct acc ttg ggt gct gcc ttc gct
gcc gaa gaa att 1296Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala
Ala Glu Glu Ile 420 425
430 gat cca aag aag aga gtt atc tta ttc att ggt gac
ggt tct ttg caa 1344Asp Pro Lys Lys Arg Val Ile Leu Phe Ile Gly Asp
Gly Ser Leu Gln 435 440
445 ttg act gtt caa gaa atc tcc acc atg atc aga tgg
ggc ttg aag cca 1392Leu Thr Val Gln Glu Ile Ser Thr Met Ile Arg Trp
Gly Leu Lys Pro 450 455 460
tac ttg ttc gta ttg aac aac gac ggt tac acc att
gaa aga ttg att 1440Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile
Glu Arg Leu Ile 465 470 475
480 cac ggt gaa acc gct caa tac aac tgt atc caa aac
tgg caa cac ttg 1488His Gly Glu Thr Ala Gln Tyr Asn Cys Ile Gln Asn
Trp Gln His Leu 485 490
495 gaa tta ttg cca act ttc ggt gcc aag gac tac gaa
gct gtc aga gtt 1536Glu Leu Leu Pro Thr Phe Gly Ala Lys Asp Tyr Glu
Ala Val Arg Val 500 505
510 tcc acc act ggt gaa tgg aac aag ttg acc act gac
gaa aag ttc caa 1584Ser Thr Thr Gly Glu Trp Asn Lys Leu Thr Thr Asp
Glu Lys Phe Gln 515 520
525 gac aac acc aga atc aga ttg atc gaa gtt atg ttg
cca act atg gat 1632Asp Asn Thr Arg Ile Arg Leu Ile Glu Val Met Leu
Pro Thr Met Asp 530 535 540
gct cca tct aac ttg gtt aag caa gct caa ttg act
gct gct acc aac 1680Ala Pro Ser Asn Leu Val Lys Gln Ala Gln Leu Thr
Ala Ala Thr Asn 545 550 555
560 gct aag aac taa
1692Ala Lys Asn
124563PRTKluyveromyces lactis 124Met Ser Glu Ile
Thr Leu Gly Arg Tyr Leu Phe Glu Arg Leu Lys Gln 1 5
10 15 Val Glu Val Gln Thr Ile Phe Gly Leu
Pro Gly Asp Phe Asn Leu Ser 20 25
30 Leu Leu Asp Asn Ile Tyr Glu Val Pro Gly Met Arg Trp Ala
Gly Asn 35 40 45
Ala Asn Glu Leu Asn Ala Ala Tyr Ala Ala Asp Gly Tyr Ala Arg Leu 50
55 60 Lys Gly Met Ser Cys
Ile Ile Thr Thr Phe Gly Val Gly Glu Leu Ser 65 70
75 80 Ala Leu Asn Gly Ile Ala Gly Ser Tyr Ala
Glu His Val Gly Val Leu 85 90
95 His Val Val Gly Val Pro Ser Val Ser Ser Gln Ala Lys Gln Leu
Leu 100 105 110 Leu
His His Thr Leu Gly Asn Gly Asp Phe Thr Val Phe His Arg Met 115
120 125 Ser Ser Asn Ile Ser Glu
Thr Thr Ala Met Ile Thr Asp Ile Asn Thr 130 135
140 Ala Pro Ala Glu Ile Asp Arg Cys Ile Arg Thr
Thr Tyr Val Ser Gln 145 150 155
160 Arg Pro Val Tyr Leu Gly Leu Pro Ala Asn Leu Val Asp Leu Thr Val
165 170 175 Pro Ala
Ser Leu Leu Asp Thr Pro Ile Asp Leu Ser Leu Lys Pro Asn 180
185 190 Asp Pro Glu Ala Glu Glu Glu
Val Ile Glu Asn Val Leu Gln Leu Ile 195 200
205 Lys Glu Ala Lys Asn Pro Val Ile Leu Ala Asp Ala
Cys Cys Ser Arg 210 215 220
His Asp Ala Lys Ala Glu Thr Lys Lys Leu Ile Asp Leu Thr Gln Phe 225
230 235 240 Pro Ala Phe
Val Thr Pro Met Gly Lys Gly Ser Ile Asp Glu Lys His 245
250 255 Pro Arg Phe Gly Gly Val Tyr Val
Gly Thr Leu Ser Ser Pro Ala Val 260 265
270 Lys Glu Ala Val Glu Ser Ala Asp Leu Val Leu Ser Val
Gly Ala Leu 275 280 285
Leu Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr Ser Tyr Lys Thr Lys 290
295 300 Asn Ile Val Glu
Phe His Ser Asp Tyr Thr Lys Ile Arg Ser Ala Thr 305 310
315 320 Phe Pro Gly Val Gln Met Lys Phe Ala
Leu Gln Lys Leu Leu Thr Lys 325 330
335 Val Ala Asp Ala Ala Lys Gly Tyr Lys Pro Val Pro Val Pro
Ser Glu 340 345 350
Pro Glu His Asn Glu Ala Val Ala Asp Ser Thr Pro Leu Lys Gln Glu
355 360 365 Trp Val Trp Thr
Gln Val Gly Glu Phe Leu Arg Glu Gly Asp Val Val 370
375 380 Ile Thr Glu Thr Gly Thr Ser Ala
Phe Gly Ile Asn Gln Thr His Phe 385 390
395 400 Pro Asn Asn Thr Tyr Gly Ile Ser Gln Val Leu Trp
Gly Ser Ile Gly 405 410
415 Phe Thr Thr Gly Ala Thr Leu Gly Ala Ala Phe Ala Ala Glu Glu Ile
420 425 430 Asp Pro Lys
Lys Arg Val Ile Leu Phe Ile Gly Asp Gly Ser Leu Gln 435
440 445 Leu Thr Val Gln Glu Ile Ser Thr
Met Ile Arg Trp Gly Leu Lys Pro 450 455
460 Tyr Leu Phe Val Leu Asn Asn Asp Gly Tyr Thr Ile Glu
Arg Leu Ile 465 470 475
480 His Gly Glu Thr Ala Gln Tyr Asn Cys Ile Gln Asn Trp Gln His Leu
485 490 495 Glu Leu Leu Pro
Thr Phe Gly Ala Lys Asp Tyr Glu Ala Val Arg Val 500
505 510 Ser Thr Thr Gly Glu Trp Asn Lys Leu
Thr Thr Asp Glu Lys Phe Gln 515 520
525 Asp Asn Thr Arg Ile Arg Leu Ile Glu Val Met Leu Pro Thr
Met Asp 530 535 540
Ala Pro Ser Asn Leu Val Lys Gln Ala Gln Leu Thr Ala Ala Thr Asn 545
550 555 560 Ala Lys Asn
1251716DNAYarrowia lipolyticaCDS(1)..(1716) 125atg agc gac tcc gaa ccc
caa atg gtc gac ctg ggc gac tat ctc ttt 48Met Ser Asp Ser Glu Pro
Gln Met Val Asp Leu Gly Asp Tyr Leu Phe 1 5
10 15 gcc cga ttc aag cag cta
ggc gtg gac tcc gtc ttt gga gtg ccc ggc 96Ala Arg Phe Lys Gln Leu
Gly Val Asp Ser Val Phe Gly Val Pro Gly 20
25 30 gac ttc aac ctc acc ctg
ttg gac cac gtg tac aat gtc gac atg cgg 144Asp Phe Asn Leu Thr Leu
Leu Asp His Val Tyr Asn Val Asp Met Arg 35
40 45 tgg gtt ggg aac aca aac
gag ctg aat gcc ggc tac tcg gcc gac ggc 192Trp Val Gly Asn Thr Asn
Glu Leu Asn Ala Gly Tyr Ser Ala Asp Gly 50
55 60 tac tcc cgg gtc aag cgg
ctg gca tgt ctt gtc acc acc ttt ggc gtg 240Tyr Ser Arg Val Lys Arg
Leu Ala Cys Leu Val Thr Thr Phe Gly Val 65 70
75 80 gga gag ctg tct gcc gtg
gct gct gtg gca ggc tcg tac gcc gag cat 288Gly Glu Leu Ser Ala Val
Ala Ala Val Ala Gly Ser Tyr Ala Glu His 85
90 95 gtg ggc gtg gtg cat gtt
gtg ggc gtt ccc agc acc tct gct gag aac 336Val Gly Val Val His Val
Val Gly Val Pro Ser Thr Ser Ala Glu Asn 100
105 110 aag cat ctg ctg ctg cac
cac aca ctc ggt aac ggc gac ttc cgg gtc 384Lys His Leu Leu Leu His
His Thr Leu Gly Asn Gly Asp Phe Arg Val 115
120 125 ttt gcc cag atg tcc aaa
ctc atc tcc gag tac acc cac cat att gag 432Phe Ala Gln Met Ser Lys
Leu Ile Ser Glu Tyr Thr His His Ile Glu 130
135 140 gac ccc agc gag gct gcc
gac gta atc gac acc gcc atc cga atc gcc 480Asp Pro Ser Glu Ala Ala
Asp Val Ile Asp Thr Ala Ile Arg Ile Ala 145 150
155 160 tac acc cac cag cgg ccc
gtt tac att gct gtg ccc tcc aac ttc tcc 528Tyr Thr His Gln Arg Pro
Val Tyr Ile Ala Val Pro Ser Asn Phe Ser 165
170 175 gag gtc gat att gcc gac
cag gct aga ctg gat acc ccc ctg gac ctt 576Glu Val Asp Ile Ala Asp
Gln Ala Arg Leu Asp Thr Pro Leu Asp Leu 180
185 190 tcg ctg cag ccc aac gac
ccc gag agc cag tac gag gtg att gag gag 624Ser Leu Gln Pro Asn Asp
Pro Glu Ser Gln Tyr Glu Val Ile Glu Glu 195
200 205 att tgc tcg cgt atc aag
gcc gcc aag aag ccc gtg att ctc gtc gac 672Ile Cys Ser Arg Ile Lys
Ala Ala Lys Lys Pro Val Ile Leu Val Asp 210
215 220 gcc tgc gct tcg cga tac
aga tgt gtg gac gag acc aag gag ctg gcc 720Ala Cys Ala Ser Arg Tyr
Arg Cys Val Asp Glu Thr Lys Glu Leu Ala 225 230
235 240 aag atc acc aac ttt gcc
tac ttt gtc act ccc atg ggt aag ggt tct 768Lys Ile Thr Asn Phe Ala
Tyr Phe Val Thr Pro Met Gly Lys Gly Ser 245
250 255 gtg gac gag gat act gac
cgg tac gga gga aca tac gtc gga tcg ctg 816Val Asp Glu Asp Thr Asp
Arg Tyr Gly Gly Thr Tyr Val Gly Ser Leu 260
265 270 act gct cct gct act gcc
gag gtg gtt gag aca gct gat ctc atc atc 864Thr Ala Pro Ala Thr Ala
Glu Val Val Glu Thr Ala Asp Leu Ile Ile 275
280 285 tcc gta gga gct ctt ctg
tcg gac ttc aac acc ggt tcc ttc tcg tac 912Ser Val Gly Ala Leu Leu
Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr 290
295 300 tcc tac tcc acc aaa aac
gtg gtg gaa ttg cat tcg gac cac gtc aaa 960Ser Tyr Ser Thr Lys Asn
Val Val Glu Leu His Ser Asp His Val Lys 305 310
315 320 atc aag tcc gcc acc tac
aac aac gtc ggc atg aaa atg ctg ttc ccg 1008Ile Lys Ser Ala Thr Tyr
Asn Asn Val Gly Met Lys Met Leu Phe Pro 325
330 335 ccc ctg ctc gaa gcc gtc
aag aaa ctg gtt gcc gag acc cct gac ttt 1056Pro Leu Leu Glu Ala Val
Lys Lys Leu Val Ala Glu Thr Pro Asp Phe 340
345 350 gca tcc aag gct ctg gct
gtt ccc gac acc act ccc aag atc ccc gag 1104Ala Ser Lys Ala Leu Ala
Val Pro Asp Thr Thr Pro Lys Ile Pro Glu 355
360 365 gta ccc gat gat cac att
acg acc cag gca tgg ctg tgg cag cgt ctc 1152Val Pro Asp Asp His Ile
Thr Thr Gln Ala Trp Leu Trp Gln Arg Leu 370
375 380 agt tac ttt ctg agg ccc
acc gac atc gtg gtc acc gag acc gga acc 1200Ser Tyr Phe Leu Arg Pro
Thr Asp Ile Val Val Thr Glu Thr Gly Thr 385 390
395 400 tcg tcc ttt gga atc atc
cag acc aag ttc ccc cac aac gtc cga ggt 1248Ser Ser Phe Gly Ile Ile
Gln Thr Lys Phe Pro His Asn Val Arg Gly 405
410 415 atc tcg cag gtg ctg tgg
ggc tct att gga tac tcg gtg gga gca gcc 1296Ile Ser Gln Val Leu Trp
Gly Ser Ile Gly Tyr Ser Val Gly Ala Ala 420
425 430 tgt gga gcc tcc att gct
gca cag gag att gac ccc cag cag cga gtg 1344Cys Gly Ala Ser Ile Ala
Ala Gln Glu Ile Asp Pro Gln Gln Arg Val 435
440 445 att ctg ttt gtg ggc gac
ggc tct ctt cag ctg acg gtg acc gag atc 1392Ile Leu Phe Val Gly Asp
Gly Ser Leu Gln Leu Thr Val Thr Glu Ile 450
455 460 tcg tgc atg atc cgc aac
aac gtc aag ccg tac att ttt gtg ctc aac 1440Ser Cys Met Ile Arg Asn
Asn Val Lys Pro Tyr Ile Phe Val Leu Asn 465 470
475 480 aac gac ggc tac acc atc
gag agg ctc att cac ggc gaa aac gcc tcg 1488Asn Asp Gly Tyr Thr Ile
Glu Arg Leu Ile His Gly Glu Asn Ala Ser 485
490 495 tac aac gat gtg cac atg
tgg aag tac tcc aag att ctc gac acg ttc 1536Tyr Asn Asp Val His Met
Trp Lys Tyr Ser Lys Ile Leu Asp Thr Phe 500
505 510 aac gcc aag gcc cac gag
tcg att gtg gtc aac acc aag ggc gag atg 1584Asn Ala Lys Ala His Glu
Ser Ile Val Val Asn Thr Lys Gly Glu Met 515
520 525 gac gct ctg ttc gac aac
gaa gag ttt gcc aag ccc gac aag atc cgg 1632Asp Ala Leu Phe Asp Asn
Glu Glu Phe Ala Lys Pro Asp Lys Ile Arg 530
535 540 ctc att gag gtc atg tgc
gac aag atg gac gcg cct gcc tcg ttg atc 1680Leu Ile Glu Val Met Cys
Asp Lys Met Asp Ala Pro Ala Ser Leu Ile 545 550
555 560 aag cag gct gag ctc tct
gcc aag acc aac gtt tag 1716Lys Gln Ala Glu Leu Ser
Ala Lys Thr Asn Val 565
570 126571PRTYarrowia
lipolytica 126Met Ser Asp Ser Glu Pro Gln Met Val Asp Leu Gly Asp Tyr Leu
Phe 1 5 10 15 Ala
Arg Phe Lys Gln Leu Gly Val Asp Ser Val Phe Gly Val Pro Gly
20 25 30 Asp Phe Asn Leu Thr
Leu Leu Asp His Val Tyr Asn Val Asp Met Arg 35
40 45 Trp Val Gly Asn Thr Asn Glu Leu Asn
Ala Gly Tyr Ser Ala Asp Gly 50 55
60 Tyr Ser Arg Val Lys Arg Leu Ala Cys Leu Val Thr Thr
Phe Gly Val 65 70 75
80 Gly Glu Leu Ser Ala Val Ala Ala Val Ala Gly Ser Tyr Ala Glu His
85 90 95 Val Gly Val Val
His Val Val Gly Val Pro Ser Thr Ser Ala Glu Asn 100
105 110 Lys His Leu Leu Leu His His Thr Leu
Gly Asn Gly Asp Phe Arg Val 115 120
125 Phe Ala Gln Met Ser Lys Leu Ile Ser Glu Tyr Thr His His
Ile Glu 130 135 140
Asp Pro Ser Glu Ala Ala Asp Val Ile Asp Thr Ala Ile Arg Ile Ala 145
150 155 160 Tyr Thr His Gln Arg
Pro Val Tyr Ile Ala Val Pro Ser Asn Phe Ser 165
170 175 Glu Val Asp Ile Ala Asp Gln Ala Arg Leu
Asp Thr Pro Leu Asp Leu 180 185
190 Ser Leu Gln Pro Asn Asp Pro Glu Ser Gln Tyr Glu Val Ile Glu
Glu 195 200 205 Ile
Cys Ser Arg Ile Lys Ala Ala Lys Lys Pro Val Ile Leu Val Asp 210
215 220 Ala Cys Ala Ser Arg Tyr
Arg Cys Val Asp Glu Thr Lys Glu Leu Ala 225 230
235 240 Lys Ile Thr Asn Phe Ala Tyr Phe Val Thr Pro
Met Gly Lys Gly Ser 245 250
255 Val Asp Glu Asp Thr Asp Arg Tyr Gly Gly Thr Tyr Val Gly Ser Leu
260 265 270 Thr Ala
Pro Ala Thr Ala Glu Val Val Glu Thr Ala Asp Leu Ile Ile 275
280 285 Ser Val Gly Ala Leu Leu Ser
Asp Phe Asn Thr Gly Ser Phe Ser Tyr 290 295
300 Ser Tyr Ser Thr Lys Asn Val Val Glu Leu His Ser
Asp His Val Lys 305 310 315
320 Ile Lys Ser Ala Thr Tyr Asn Asn Val Gly Met Lys Met Leu Phe Pro
325 330 335 Pro Leu Leu
Glu Ala Val Lys Lys Leu Val Ala Glu Thr Pro Asp Phe 340
345 350 Ala Ser Lys Ala Leu Ala Val Pro
Asp Thr Thr Pro Lys Ile Pro Glu 355 360
365 Val Pro Asp Asp His Ile Thr Thr Gln Ala Trp Leu Trp
Gln Arg Leu 370 375 380
Ser Tyr Phe Leu Arg Pro Thr Asp Ile Val Val Thr Glu Thr Gly Thr 385
390 395 400 Ser Ser Phe Gly
Ile Ile Gln Thr Lys Phe Pro His Asn Val Arg Gly 405
410 415 Ile Ser Gln Val Leu Trp Gly Ser Ile
Gly Tyr Ser Val Gly Ala Ala 420 425
430 Cys Gly Ala Ser Ile Ala Ala Gln Glu Ile Asp Pro Gln Gln
Arg Val 435 440 445
Ile Leu Phe Val Gly Asp Gly Ser Leu Gln Leu Thr Val Thr Glu Ile 450
455 460 Ser Cys Met Ile Arg
Asn Asn Val Lys Pro Tyr Ile Phe Val Leu Asn 465 470
475 480 Asn Asp Gly Tyr Thr Ile Glu Arg Leu Ile
His Gly Glu Asn Ala Ser 485 490
495 Tyr Asn Asp Val His Met Trp Lys Tyr Ser Lys Ile Leu Asp Thr
Phe 500 505 510 Asn
Ala Lys Ala His Glu Ser Ile Val Val Asn Thr Lys Gly Glu Met 515
520 525 Asp Ala Leu Phe Asp Asn
Glu Glu Phe Ala Lys Pro Asp Lys Ile Arg 530 535
540 Leu Ile Glu Val Met Cys Asp Lys Met Asp Ala
Pro Ala Ser Leu Ile 545 550 555
560 Lys Gln Ala Glu Leu Ser Ala Lys Thr Asn Val 565
570 1271716DNASchizosaccharomyces pombeCDS(1)..(1716)
127atg agt ggg gat att tta gtc ggt gaa tat cta ttc aaa agg ctt gaa
48Met Ser Gly Asp Ile Leu Val Gly Glu Tyr Leu Phe Lys Arg Leu Glu
1 5 10 15
caa tta ggg gtc aag tcc att ctt ggt gtt cca gga gat ttc aat tta
96Gln Leu Gly Val Lys Ser Ile Leu Gly Val Pro Gly Asp Phe Asn Leu
20 25 30
gct cta ctt gac tta att gag aaa gtt gga gat gag aaa ttt cgt tgg
144Ala Leu Leu Asp Leu Ile Glu Lys Val Gly Asp Glu Lys Phe Arg Trp
35 40 45
gtt ggc aat acc aat gag ttg aat ggt gct tat gcc gct gat ggt tat
192Val Gly Asn Thr Asn Glu Leu Asn Gly Ala Tyr Ala Ala Asp Gly Tyr
50 55 60
gct cgt gtt aat ggt ctt tca gcc att gtt aca acg ttc ggc gtg gga
240Ala Arg Val Asn Gly Leu Ser Ala Ile Val Thr Thr Phe Gly Val Gly
65 70 75 80
gag ctt tcc gct att aat gga gtg gca ggt tct tat gcg gag cat gtc
288Glu Leu Ser Ala Ile Asn Gly Val Ala Gly Ser Tyr Ala Glu His Val
85 90 95
cca gta gtt cat att gtt gga atg cct tcc aca aag gtg caa gat act
336Pro Val Val His Ile Val Gly Met Pro Ser Thr Lys Val Gln Asp Thr
100 105 110
gga gct ttg ctt cat cat act tta gga gat gga gac ttt cgc act ttc
384Gly Ala Leu Leu His His Thr Leu Gly Asp Gly Asp Phe Arg Thr Phe
115 120 125
atg gat atg ttt aag aaa gtt tct gcc tac agt ata atg atc gat aac
432Met Asp Met Phe Lys Lys Val Ser Ala Tyr Ser Ile Met Ile Asp Asn
130 135 140
gga aac gat gca gct gaa aag atc gat gaa gcc ttg tcg att tgt tat
480Gly Asn Asp Ala Ala Glu Lys Ile Asp Glu Ala Leu Ser Ile Cys Tyr
145 150 155 160
aaa aag gct agg cct gtt tac att ggt att cct tct gat gct ggc tac
528Lys Lys Ala Arg Pro Val Tyr Ile Gly Ile Pro Ser Asp Ala Gly Tyr
165 170 175
ttc aaa gca tct tca tca aat ctt ggg aaa aga cta aag ctc gag gag
576Phe Lys Ala Ser Ser Ser Asn Leu Gly Lys Arg Leu Lys Leu Glu Glu
180 185 190
gat act aac gat cca gca gtt gag caa gaa gtc atc aat cat atc tcg
624Asp Thr Asn Asp Pro Ala Val Glu Gln Glu Val Ile Asn His Ile Ser
195 200 205
gaa atg gtt gtc aat gca aag aaa cca gtg att tta att gac gct tgt
672Glu Met Val Val Asn Ala Lys Lys Pro Val Ile Leu Ile Asp Ala Cys
210 215 220
gct gta aga cat cgt gtc gtt cca gaa gta cat gag ctg att aaa ttg
720Ala Val Arg His Arg Val Val Pro Glu Val His Glu Leu Ile Lys Leu
225 230 235 240
acc cat ttc cct aca tat gta act ccc atg ggt aaa tct gca att gac
768Thr His Phe Pro Thr Tyr Val Thr Pro Met Gly Lys Ser Ala Ile Asp
245 250 255
gaa act tcg caa ttt ttt gac ggc gtt tat gtt ggt tca att tca gat
816Glu Thr Ser Gln Phe Phe Asp Gly Val Tyr Val Gly Ser Ile Ser Asp
260 265 270
cct gaa gtt aaa gac aga att gaa tcc act gat ctg ttg cta tcc atc
864Pro Glu Val Lys Asp Arg Ile Glu Ser Thr Asp Leu Leu Leu Ser Ile
275 280 285
ggt gct ctc aaa tca gac ttt aac acg ggt tcc ttc tct tac cac ctc
912Gly Ala Leu Lys Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr His Leu
290 295 300
agc caa aag aat gcc gtt gag ttt cat tca gac cac atg cgc att cga
960Ser Gln Lys Asn Ala Val Glu Phe His Ser Asp His Met Arg Ile Arg
305 310 315 320
tat gct ctt tat cca aat gta gcc atg aag tat att ctt cgc aaa ctg
1008Tyr Ala Leu Tyr Pro Asn Val Ala Met Lys Tyr Ile Leu Arg Lys Leu
325 330 335
ttg aaa gta ctt gat gct tct atg tgt cat tcc aag gct gct cct acc
1056Leu Lys Val Leu Asp Ala Ser Met Cys His Ser Lys Ala Ala Pro Thr
340 345 350
att ggc tac aac atc aag cct aag cat gcg gaa gga tat tct tcc aac
1104Ile Gly Tyr Asn Ile Lys Pro Lys His Ala Glu Gly Tyr Ser Ser Asn
355 360 365
gag att act cat tgc tgg ttt tgg cct aaa ttt agt gaa ttt ttg aag
1152Glu Ile Thr His Cys Trp Phe Trp Pro Lys Phe Ser Glu Phe Leu Lys
370 375 380
ccc cga gat gtt ttg atc acc gag act gga act gca aac ttt ggt gtc
1200Pro Arg Asp Val Leu Ile Thr Glu Thr Gly Thr Ala Asn Phe Gly Val
385 390 395 400
ctt gat tgc agg ttt cca aag gat gta aca gcc att tcc cag gta tta
1248Leu Asp Cys Arg Phe Pro Lys Asp Val Thr Ala Ile Ser Gln Val Leu
405 410 415
tgg gga tct att gga tac tcc gtt ggt gca atg ttt ggt gct gtt ttg
1296Trp Gly Ser Ile Gly Tyr Ser Val Gly Ala Met Phe Gly Ala Val Leu
420 425 430
gcc gtc cac gat tct aaa gag ccc gat cgt cgt acc att ctt gta gta
1344Ala Val His Asp Ser Lys Glu Pro Asp Arg Arg Thr Ile Leu Val Val
435 440 445
ggt gat gga tcc tta caa ctg acg att aca gag att tca acc tgc att
1392Gly Asp Gly Ser Leu Gln Leu Thr Ile Thr Glu Ile Ser Thr Cys Ile
450 455 460
cgc cat aac ctc aaa cca att att ttc ata att aac aac gac ggt tac
1440Arg His Asn Leu Lys Pro Ile Ile Phe Ile Ile Asn Asn Asp Gly Tyr
465 470 475 480
acc att gag cgt tta att cat ggt ttg cat gct agc tat aac gaa att
1488Thr Ile Glu Arg Leu Ile His Gly Leu His Ala Ser Tyr Asn Glu Ile
485 490 495
aac act aaa tgg ggc tac caa cag att ccc aag ttt ttc gga gct gct
1536Asn Thr Lys Trp Gly Tyr Gln Gln Ile Pro Lys Phe Phe Gly Ala Ala
500 505 510
gaa aac cac ttc cgc act tac tgt gtt aaa act cct act gac gtt gaa
1584Glu Asn His Phe Arg Thr Tyr Cys Val Lys Thr Pro Thr Asp Val Glu
515 520 525
aag ttg ttt agc gac aag gag ttt gca aat gca gat gtc att caa gta
1632Lys Leu Phe Ser Asp Lys Glu Phe Ala Asn Ala Asp Val Ile Gln Val
530 535 540
gtt gag ctt gta atg cct atg ttg gat gca cct cgt gtc cta gtt gag
1680Val Glu Leu Val Met Pro Met Leu Asp Ala Pro Arg Val Leu Val Glu
545 550 555 560
caa gcc aag ttg acg tct aag atc aat aag caa tga
1716Gln Ala Lys Leu Thr Ser Lys Ile Asn Lys Gln
565 570
128571PRTSchizosaccharomyces pombe 128Met Ser Gly Asp Ile Leu Val Gly
Glu Tyr Leu Phe Lys Arg Leu Glu 1 5 10
15 Gln Leu Gly Val Lys Ser Ile Leu Gly Val Pro Gly Asp
Phe Asn Leu 20 25 30
Ala Leu Leu Asp Leu Ile Glu Lys Val Gly Asp Glu Lys Phe Arg Trp
35 40 45 Val Gly Asn Thr
Asn Glu Leu Asn Gly Ala Tyr Ala Ala Asp Gly Tyr 50
55 60 Ala Arg Val Asn Gly Leu Ser Ala
Ile Val Thr Thr Phe Gly Val Gly 65 70
75 80 Glu Leu Ser Ala Ile Asn Gly Val Ala Gly Ser Tyr
Ala Glu His Val 85 90
95 Pro Val Val His Ile Val Gly Met Pro Ser Thr Lys Val Gln Asp Thr
100 105 110 Gly Ala Leu
Leu His His Thr Leu Gly Asp Gly Asp Phe Arg Thr Phe 115
120 125 Met Asp Met Phe Lys Lys Val Ser
Ala Tyr Ser Ile Met Ile Asp Asn 130 135
140 Gly Asn Asp Ala Ala Glu Lys Ile Asp Glu Ala Leu Ser
Ile Cys Tyr 145 150 155
160 Lys Lys Ala Arg Pro Val Tyr Ile Gly Ile Pro Ser Asp Ala Gly Tyr
165 170 175 Phe Lys Ala Ser
Ser Ser Asn Leu Gly Lys Arg Leu Lys Leu Glu Glu 180
185 190 Asp Thr Asn Asp Pro Ala Val Glu Gln
Glu Val Ile Asn His Ile Ser 195 200
205 Glu Met Val Val Asn Ala Lys Lys Pro Val Ile Leu Ile Asp
Ala Cys 210 215 220
Ala Val Arg His Arg Val Val Pro Glu Val His Glu Leu Ile Lys Leu 225
230 235 240 Thr His Phe Pro Thr
Tyr Val Thr Pro Met Gly Lys Ser Ala Ile Asp 245
250 255 Glu Thr Ser Gln Phe Phe Asp Gly Val Tyr
Val Gly Ser Ile Ser Asp 260 265
270 Pro Glu Val Lys Asp Arg Ile Glu Ser Thr Asp Leu Leu Leu Ser
Ile 275 280 285 Gly
Ala Leu Lys Ser Asp Phe Asn Thr Gly Ser Phe Ser Tyr His Leu 290
295 300 Ser Gln Lys Asn Ala Val
Glu Phe His Ser Asp His Met Arg Ile Arg 305 310
315 320 Tyr Ala Leu Tyr Pro Asn Val Ala Met Lys Tyr
Ile Leu Arg Lys Leu 325 330
335 Leu Lys Val Leu Asp Ala Ser Met Cys His Ser Lys Ala Ala Pro Thr
340 345 350 Ile Gly
Tyr Asn Ile Lys Pro Lys His Ala Glu Gly Tyr Ser Ser Asn 355
360 365 Glu Ile Thr His Cys Trp Phe
Trp Pro Lys Phe Ser Glu Phe Leu Lys 370 375
380 Pro Arg Asp Val Leu Ile Thr Glu Thr Gly Thr Ala
Asn Phe Gly Val 385 390 395
400 Leu Asp Cys Arg Phe Pro Lys Asp Val Thr Ala Ile Ser Gln Val Leu
405 410 415 Trp Gly Ser
Ile Gly Tyr Ser Val Gly Ala Met Phe Gly Ala Val Leu 420
425 430 Ala Val His Asp Ser Lys Glu Pro
Asp Arg Arg Thr Ile Leu Val Val 435 440
445 Gly Asp Gly Ser Leu Gln Leu Thr Ile Thr Glu Ile Ser
Thr Cys Ile 450 455 460
Arg His Asn Leu Lys Pro Ile Ile Phe Ile Ile Asn Asn Asp Gly Tyr 465
470 475 480 Thr Ile Glu Arg
Leu Ile His Gly Leu His Ala Ser Tyr Asn Glu Ile 485
490 495 Asn Thr Lys Trp Gly Tyr Gln Gln Ile
Pro Lys Phe Phe Gly Ala Ala 500 505
510 Glu Asn His Phe Arg Thr Tyr Cys Val Lys Thr Pro Thr Asp
Val Glu 515 520 525
Lys Leu Phe Ser Asp Lys Glu Phe Ala Asn Ala Asp Val Ile Gln Val 530
535 540 Val Glu Leu Val Met
Pro Met Leu Asp Ala Pro Arg Val Leu Val Glu 545 550
555 560 Gln Ala Lys Leu Thr Ser Lys Ile Asn Lys
Gln 565 570
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