Microorganism producing citric acid and its construction method and application

Abstract

The present invention discloses a microorganism for producing citric acid, a construction method thereof and an application thereof. Based on the genome data mining of Aspergillus niger M202 strain, the present invention unexpectedly found that the inactivation of four proteins can increase the yield of citric acid in Aspergillus niger. Further analysis found that the coding genes of these four proteins belong to a gene cluster with unknown function, and the inactivation of the gene cluster can also promote the production of citric acid, significantly improving the acid production level of the strain. In addition, the activity of the citric acid transporter in the production strain is enhanced, resulting in enhanced citric acid efflux, further improving the production of citric acid. The present invention can provide new ideas and targets for those skilled in the art to construct citric acid production strains, and provide reference for constructing production strains with higher citric acid production.

Description

Technical Field

[0001] The invention belongs to the field of molecular biology and bioengineering, and specifically relates to the application of endogenous polypeptides derived from Aspergillus niger in the production of citric acid, a strain for producing citric acid, and a construction method and application of the strain. Background Art

[0002] Citric acid is a very important organic acid, a natural preservative and food additive, widely used in industry, food and cosmetics. It is mainly produced by microbial fermentation, and its main production strains include yeast and Aspergillus niger. Aspergillus niger is the most competitive industrial strain in the citric acid industry. The global annual production of citric acid produced by Aspergillus niger reaches 2 million tons, with an output value of more than 2 billion US dollars, and it is increasing at an annual rate of 5%.

[0003] Due to the difficulty of genetic modification of Aspergillus niger, early research mainly focused on the optimization of fermentation process to increase the yield of Aspergillus niger, such as the continuous optimization of fermentation conditions such as carbon source, nitrogen source, pH, etc. In recent years, with the development of metabolic engineering and the continuous development of Aspergillus niger genetic manipulation tools, reports on increasing citric acid production through genetic engineering have also appeared one after another. The targets of the modification include increasing the supply of its substrate acetyl-CoA, increasing the efflux of citric acid, increasing respiratory chain regulation, etc. These genetic modifications have increased the yield of citric acid in Aspergillus niger to varying degrees. However, with the growing demand for citric acid in the global market, how to find new modification targets, construct efficient Aspergillus niger engineered strains to increase the yield of citric acid, and further reduce production costs are problems that need to be solved in this field. Summary of the invention

[0004] In order to overcome the deficiencies in the prior art, the present invention is based on the genome data mining of Aspergillus niger M202 strain, and unexpectedly found that the inactivation of four proteins can increase the production of citric acid in Aspergillus niger. Further analysis found that the coding genes of these four proteins belong to a gene cluster with unknown function, and the inactivation of this gene cluster can also promote the production of citric acid, significantly improve the acid production level of the strain, and in addition, the activity of the citric acid transporter CexA in the production strain is enhanced, resulting in enhanced citric acid efflux, further improving the production of citric acid. Therefore, the primary purpose of the present invention is to provide a use of an endogenous active polypeptide in the production of citric acid.

[0005] Another object of the present invention is to provide a citric acid producing strain.

[0006] Another object of the present invention is to provide a method for producing citric acid.

[0007] Another object of the present invention is to provide a method for constructing a citric acid producing strain.

[0008] In order to achieve the above object, the present invention adopts the following technical solution:

[0009] In the first aspect of the present invention, there is provided a use of an endogenous active polypeptide in producing citric acid by microbial fermentation, wherein the polypeptide is:

[0010] (A) the amino acid sequence is shown in any one of SEQ ID NOs: 1-4; or

[0011] (B) has more than 98% homology with the amino acid sequence of the protein described in (A), and is derived from Aspergillus niger and has the same activity as the protein described in (A).

[0012] Preferably, the amino acid sequence of the endogenous active polypeptide is shown in any one of SEQ ID NOs: 1-4.

[0013] Preferably, in the above use, the activity of one endogenous active polypeptide in (A) or (B) is weakened or completely eliminated.

[0014] In a second aspect of the present invention, there is provided a use of a gene cluster in producing citric acid by microbial fermentation, wherein the gene cluster is:

[0015] (A) the nucleotide sequence is shown in SEQ ID NO:5; or

[0016] (B) has more than 98% homology with the nucleotide sequence of (A) and is derived from Aspergillus niger.

[0017] Preferably, the nucleotide sequence of the gene cluster is shown in SEQ ID NO:5.

[0018] Preferably, in the above use, the entire gene cluster in (A) or (B) is inactivated.

[0019] In a third aspect of the present invention, a citric acid producing strain is provided, wherein the strain is modified:

[0020] 1) weakening or completely removing the activity of an endogenous active polypeptide, wherein the endogenous active polypeptide is selected from:

[0021] (A) the amino acid sequence is shown in any one of SEQ ID NOs: 1-4; or

[0022] (B) has an amino acid sequence homology of more than 98% with the protein described in (A), and is derived from Aspergillus niger and has the same activity as the protein described in (A);

[0023] or,

[0024] 2) Inactivating a gene cluster, the gene cluster being:

[0025] (A) the nucleotide sequence is shown in SEQ ID NO:5; or

[0026] (B) has more than 98% homology with the nucleotide sequence of (A) and is derived from Aspergillus niger.

[0027] Preferably, the activity reduction or complete removal of the endogenous polypeptide can be achieved by one or a combination of the following methods: partial or complete knockout of the polypeptide encoding gene; mutation of the encoding gene; changing the promoter, translation regulatory region or coding region codon of the encoding gene to weaken its transcription or translation; changing the coding gene sequence to weaken its mRNA stability or destabilize the structure of the encoded protein; or any other method of inactivating the gene by modifying the gene coding region and its adjacent upstream and downstream regions.

[0028] In a preferred embodiment, the gene coding sequence is subjected to frameshift mutation, missense mutation, deletion, start codon change, etc.

[0029] Further preferably, the activity of the endogenous active polypeptide in the strain is weakened relative to that of the unmodified endogenous active polypeptide, which means that the transcription and expression of the encoding gene of the endogenous active polypeptide is reduced by at least 30%, preferably at least 40%, more preferably at least 50%, for example, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or completely disappears; or the activity of the endogenous active polypeptide is reduced by at least 30%, preferably at least 40%, more preferably at least 50%, for example, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or completely disappears.

[0030] Furthermore, the activity of the citrate transporter in the production strain is enhanced.

[0031] Preferably, the citrate transporter is a polypeptide having an amino acid sequence as shown in SEQ ID NO:6.

[0032] Preferably, the citric acid producing strain includes but is not limited to any one of Aspergilus niger, Aspergillus nidulans, Aspergillus oryzae, Penicillium chrysogenum, Trichoderma reesei, Ustilagomaydis, Myceliophthora thermophila and the like, preferably Aspergilus niger, Aspergillus nidulans, Aspergillus oryzae, Trichoderma reesei, and most preferably Aspergilus niger.

[0033] In a specific embodiment of the present invention, the endogenous active polypeptide is an amino acid sequence as shown in any one of SEQ ID NOs: 1-4, or the nucleotide sequence of the gene cluster is as shown in SEQ ID NO: 5.

[0034] In a fourth aspect of the present invention, a method for constructing a citric acid producing strain is provided, the method comprising genetically modifying:

[0035] 1) weakening or completely removing the activity of an endogenous active polypeptide, wherein the endogenous active polypeptide is selected from:

[0036] (A) the amino acid sequence is shown in any one of SEQ ID NOs: 1-4; or

[0037] (B) has an amino acid sequence homology of more than 98% with the protein described in (A), and is derived from Aspergillus niger and has the same activity as the protein described in (A);

[0038] or,

[0039] 2) Inactivating a gene cluster, the gene cluster being:

[0040] (A) the nucleotide sequence is shown in SEQ ID NO:5; or

[0041] (B) has more than 98% homology with the nucleotide sequence of (A) and is derived from Aspergillus niger.

[0042] Preferably, the activity reduction or complete removal of the endogenous polypeptide can be achieved by one or a combination of the following methods: partial or complete knockout of the polypeptide encoding gene; mutation of the encoding gene; changing the promoter, translation regulatory region or coding region codon of the encoding gene to weaken its transcription or translation; changing the coding gene sequence to weaken its mRNA stability or destabilize the structure of the encoded protein; or any other method of inactivating the gene by modifying the gene coding region and its adjacent upstream and downstream regions.

[0043] In a preferred embodiment, the gene coding sequence is subjected to frameshift mutation, missense mutation, deletion, start codon change, etc.

[0044] Further preferably, in the above construction method, the activity of the endogenous active polypeptide in the strain is weakened relative to that of the unmodified protein, which means that the transcription and expression of the encoding gene of the endogenous active polypeptide is reduced by at least 30%, preferably at least 40%, more preferably at least 50%, for example, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or completely disappears; or the activity of the endogenous active polypeptide is reduced by at least 30%, preferably at least 40%, more preferably at least 50%, for example, at least 60%, at least 70%, at least 80%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or completely disappears.

[0045] Furthermore, the activity of the citrate transporter in the production strain is enhanced.

[0046] Preferably, the citrate transporter is a polypeptide having an amino acid sequence as shown in SEQ ID NO:6.

[0047] Preferably, the citric acid producing strain includes but is not limited to any one of Aspergilus niger, Aspergillus nidulans, Aspergillus oryzae, Penicillium chrysogenum, Trichoderma reesei, Ustilagomaydis, Myceliophthora thermophila and the like, preferably Aspergilus niger, Aspergillus nidulans, Aspergillus oryzae, Trichoderma reesei, and most preferably Aspergilus niger.

[0048] In a specific embodiment of the present invention, the endogenous active polypeptide is an amino acid sequence as shown in any one of SEQ ID NOs: 1-4, or the nucleotide sequence of the gene cluster is as shown in SEQ ID NO: 5.

[0049] In a fifth aspect of the present invention, a method for producing citric acid is provided, the method comprising: (1) culturing the citric acid-producing strain of the third aspect, or the citric acid-producing strain constructed in the fourth aspect, to produce citric acid; and

[0050] (2) Optionally separating citric acid from step (1).

[0051] In the sixth aspect of the present invention, there is provided use of the citric acid producing strain of the third aspect or the citric acid producing strain constructed of the fourth aspect in producing citric acid.

[0052] Beneficial effects of the present invention

[0053] 1. The present invention is based on the genome data mining of Aspergillus niger M202 strain. It is unexpectedly found that the inactivation of 4 proteins can improve the output of Aspergillus niger citric acid. Further analysis shows that the coding genes of these 4 proteins belong to a gene cluster with unknown function, and the inactivation of this gene cluster can also promote the production of citric acid, significantly improve the acid production level of the strain, and compared with the wild-type strain containing endogenous polypeptides, the citric acid production of the citric acid production strain can be increased by 30%. In addition, the activity of the citric acid transporter in the production strain is enhanced, resulting in the strengthening of citric acid efflux, further promoting the production of citric acid, and the citric acid production increase range is between 2.01-2.30 times. Therefore, it can be used for microbial fermentation to produce citric acid in practice, and the production cost of citric acid can be further reduced, with good industrial application potential.

[0054] 2. The present invention has found through screening that the inactivation of a gene cluster with unknown function can increase the production of citric acid in the engineered strain, providing a new idea for the genetic modification of Aspergillus niger. DETAILED DESCRIPTION

[0055] Definition of terms

[0056] The "gene cluster" used in the present invention specifically refers to a gene cluster derived from Aspergillus niger as shown in SEQ ID NO: 5, whose function is unknown. The four proteins in the gene cluster are a putative non-ribosomal peptide synthase (NRPS), a fatty acid synthesis-related protein (FAS), a putative polyketide synthase (PKS1), and a putative polyketide synthase (PKS2), which have the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, and SEQ ID NO: 4, respectively.

[0057] As used herein, "citrate transporter" refers to an efflux protein that can transport intracellular citrate to the extracellular space. For example, the citrate efflux protein of the present invention has the amino acid sequence of SEQ ID NO: 6, or a derivative protein or polypeptide formed by replacing, deleting or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO: 6 and still having the function of a citrate efflux protein.

[0058] The terms "polypeptide", "active polypeptide" and "protein" of the present invention are used interchangeably herein and are amino acid polymers of any length. The polymer may be linear or branched, it may contain modified amino acids, and it may be interrupted by non-amino acids. The term also includes amino acid polymers that have been modified (e.g., disulfide bond formation, glycosylation, lipidation, acetylation, phosphorylation or any other operation, such as conjugation with a labeling component).

[0059] The term "expression" of the present invention includes any step involved in the production of the polypeptide including, but not limited to, transcription, post-transcriptional modification, translation, post-translational modification, and secretion.

[0060] The term "exogenous" used in the present invention refers to the inclusion of substances that did not originally exist in a system. For example, if a coding gene that did not originally exist in a strain is introduced into the strain through transformation, the gene is "exogenous" to the strain.

[0061] As used herein, the terms "unmodified," "wild type," "endogenous," and "naturally occurring" refer to an object that can be found in nature. For example, a polypeptide or polynucleotide sequence that is present in an organism, can be isolated from a source in nature, and has not been intentionally modified by humans in the laboratory is naturally occurring.

[0062] The term "native state" as used herein refers to the activity of a polypeptide in an unmodified state in a microorganism, ie, the activity in the natural state.

[0063] In some embodiments, the wild-type strain of the present invention refers to a strain comprising an endogenous polypeptide, wherein the endogenous polypeptide is a polypeptide having an amino acid sequence as shown in any one of SEQ ID NO: 1 to SEQ ID NO: 4, or a polypeptide having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identity with any one of SEQ ID NO: 1 to SEQ ID NO: 4; or, the wild-type strain of the present invention refers to a strain comprising a gene cluster, wherein the gene cluster is:

[0064] (A) the nucleotide sequence is shown in SEQ ID NO:5; or

[0065] (B) has more than 98% homology with the nucleotide sequence of (A) and is derived from Aspergillus niger.

[0066] The "polynucleotide" mentioned in the present invention generally refers to polyribonucleotides and polydeoxyribonucleotides, which can be unmodified RNA or DNA, or modified RNA or DNA.

[0067] "Sequence homology" and "identity percentage" of the present invention refer to the percentage of nucleotides or amino acids that are identical (i.e., identical) between two or more polynucleotides or polypeptides. The sequence identity between two or more polynucleotides or polypeptides can be determined by the following method: the nucleotide or amino acid sequences of the polynucleotides or polypeptides are aligned and the number of positions containing the same nucleotide or amino acid residue in the aligned polynucleotides or polypeptides is scored, and compared with the number of positions containing different nucleotides or amino acid residues in the aligned polynucleotides or polypeptides. The polynucleotides may differ at one position, for example, by containing different nucleotides (i.e., substitutions or mutations) or missing nucleotides (i.e., nucleotide insertions or nucleotide deletions in one or two polynucleotides). The polypeptides may differ at one position, for example, by containing different amino acids (i.e., substitutions or mutations) or missing amino acids (i.e., amino acid insertions or amino acid deletions in one or two polypeptides). Sequence identity can be calculated by dividing the number of positions containing the same nucleotide or amino acid residue by the total number of amino acid residues in the polynucleotide or polypeptide. For example, the identity percentage can be calculated by dividing the number of positions containing the same nucleotide or amino acid residue by the total number of nucleotides or amino acid residues in the polynucleotide or polypeptide and multiplying by 100.

[0068] Those skilled in the art know that it is more important to find sites that can achieve the desired purpose when mutating wild-type polypeptides to enhance activity. Therefore, based on the teachings of the present invention, those skilled in the art will mutate the amino acid sequence shown in sequence 1 or 2, and detect the relevant activity of the mutant. In addition, it is not difficult for those of ordinary skill in the art to know that changing a few amino acid residues in certain regions of the polypeptide, such as non-important regions, will not substantially change the biological activity. For example, the sequence obtained by appropriately replacing certain amino acids will not affect its activity (see Watson et al., Molecular Biology of The Gene, Fourth Edition, 1987, The Benjamin / Cummings Pub. Co. P224). Therefore, those of ordinary skill in the art can implement such replacements and ensure that the resulting molecules still have the desired biological activity.

[0069] In a specific embodiment, the wild-type strain is Aspergillus niger containing endogenous polypeptides. The "modification" described in the present invention refers to any genetic manipulation performed on the wild-type strain, including but not limited to various molecular biological methods.

[0070] The term "reduced activity or complete elimination" in the present invention means that the intracellular activity of a protein in a microorganism is reduced, weakened or even completely eliminated compared with the activity of the protein in its natural state through modification. This includes not only the effect lower than the original function due to the reduction of the activity of the protein itself, but also the generation of genes that are not expressed, or the expression product that is expressed but has no activity or has reduced activity.

[0071] The term "weakening or complete removal of endogenous polypeptide activity" in the present invention can be achieved by modification, including but not limited to the following methods: deleting part or all of the coding gene, gene reading frame shift mutation, weakening the transcription or translation strength, or using genes or alleles encoding corresponding enzymes or proteins with lower activity, or making the corresponding genes or enzymes inactive, and optionally using these methods in combination. The reduction of gene expression can be achieved by adopting appropriate culture methods or genetic modification (mutation) of the signal structure of gene expression, for example, the signal structure of gene expression is a repressor gene, an active gene, an operator, a promoter, a weakener, a ribosome binding site, a start codon and a terminator. The terms "weakening or complete removal", "reduction", "disappearance" and "inactivation" of the present invention are interchangeable.

[0072] The term "expression control sequence" of the present invention, i.e., the nucleotide sequence that regulates gene expression, refers to a fragment that can increase or decrease the expression of a specific gene in an individual, including but not limited to promoters, transcription factor binding sites, ribosome binding sites, sequences for transcription and translation termination during regulation, etc.

[0073] The "citric acid producing strain" of the present invention has the meaning generally understood by those of ordinary skill in the art, and refers to a strain that can produce citric acid and accumulate citric acid when cultivated in a culture medium, or can secrete citric acid into a culture medium, that is, can obtain free citric acid outside the cell. For example, it can be a naturally occurring citric acid producing strain, or it can be a citric acid producing engineered strain obtained through genetic modification. The citric acid producing strain of the present invention can be a eukaryotic cell, including but not limited to Aspergillus niger, Aspergillus nidulans, Aspergillus oryzae, Penicillium chrysogenum, Trichodermareesei, Ustilago maydis, Myceliophthora thermophila and other strains, preferably Aspergillus niger, Aspergillus nidulans, Aspergillus oryzae, Trichodermareesei, most preferably Aspergillus niger, and a mutant or strain producing citric acid prepared by the above-mentioned strain.

[0074] The term "transformation" of the present invention has the meaning generally understood by those skilled in the art, i.e., the process of introducing exogenous DNA into a host. The method of transformation includes any method of introducing nucleic acid into a cell, including but not limited to electroporation, calcium phosphate (CaPO4) precipitation, calcium chloride (CaCl2) precipitation, microinjection, polyethylene glycol (PEG) method, DEAE-dextran method, cationic liposome method and lithium acetate-DMSO method.

[0075] In the present invention, the culture of the strain can be carried out according to conventional methods in the art, including but not limited to well plate culture, shake flask culture, batch culture, continuous culture and batch fed culture, and various culture conditions such as temperature, time and pH value of the culture medium can be appropriately adjusted according to actual conditions.

[0076] As used herein, the terms "comprising," "having," "including," or "containing" are intended to be inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0077] As used herein, "about" means that a value includes the standard deviation of error for the device or method being employed to determine the value.

[0078] As used herein, the definition of "or" is only alternatives and "and / or", but unless it is explicitly stated that there are only alternatives or the alternatives are mutually exclusive, the term "or" in a claim means "and / or".

[0079] The selected / optional / preferred "numerical range" used herein includes both the numerical endpoints at both ends of the range and all the natural numbers covered between the numerical endpoints relative to the aforementioned numerical endpoints.

[0080] The technical scheme of the present invention will be further described in detail below in conjunction with specific embodiments. It should be understood that the following embodiments are only exemplary descriptions and explanations of the present invention and should not be construed as limiting the scope of protection of the present invention. All technologies implemented based on the above content of the present invention are included in the scope that the present invention is intended to protect.

[0081] Unless otherwise specified, the raw materials and reagents used in the following examples are all commercially available products or can be prepared by known methods. The experimental methods in the following examples without specifying specific conditions are usually carried out according to conventional conditions such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 4th edition), or according to the conditions recommended by the manufacturer.

[0082] Example 1. Construction of sgRNA expression plasmid for genes with unknown functions

[0083] Based on the genome data mining of Aspergillus niger M202 strain (purchased from the strain resource library of Shanghai Institute of Microorganisms and Engineering Technology Co., Ltd., with public accession number M202), the present invention predicts the functions of four unknown genes, and predicts the protein shown in SEQ ID NO:1 as a non-ribosomal peptide synthase (NRPS), the protein shown in SEQ ID NO:2 as a fatty acid synthesis-related protein (FAS), the protein shown in SEQ ID NO:3 as a polyketide synthase (PKS1), and the protein shown in SEQ ID NO:4 as a polyketide synthase (PKS2). The specific amino acid sequences are shown in SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4, respectively.

[0084] Using the genome SEQ ID NO: 5 of Aspergillus niger M202 as a template, the upstream and downstream primers for the construction of the sgRNA expression cassette were designed, and the 5S rRNA of Aspergillus niger itself was used as a promoter, and the GoldenGate method was used to construct the sgRNA expression plasmid. The GoldenGate reaction product was transformed into Escherichia coli Trans-T1, and the corresponding recombinant plasmid was extracted. After restriction digestion and sequencing verification, the expression plasmids pNRPS-sgRNA1, pNRPS-sgRNA2, pFAS-sgRNA1, pFAS-sgRNA, pPKS1-sgRNA1, pPKS1-sgRNA2, pPKS2-sgRNA1, pPKS2-sgRNA2 of each protein sgRNA were obtained. The specific primer sequences are shown in Table 1.

[0085] Table 1 Primers for construction of sgRNA expression plasmid

[0086]

[0087] Example 2. Construction of gene-inactivated strains

[0088] The Cas9 protein expression vector pCas9 (see Zhang L, et al. Disruption or reduced expression of the orotidine-5'-decarboxylase gene pyrG increases citric acid production: a new discovery during recyclable genome editing in Aspergillus niger. Microb Cell Fact. 2020Mar24; 19(1):76), the sgRNA expression cassette fragment and the donor DNA fragment were co-transformed into the protoplast cells of Aspergillus niger M202 to construct strains with inactivated genes. The specific operations are as follows:

[0089] (1) Preparation of sgRNA expression cassette fragment

[0090] Using 5S-Fm (SEQ ID NO: 23: GGTTGGAGATTCCAGACTCAG) and sgRNA-Rm (SEQ ID NO: 24: AAAAAAGCACCGACTCGGTGCCAC) as primers, PCR amplification was performed using the sgRNA expression plasmid constructed in Example 1 as a template to obtain the sgRNA expression cassette fragments NRPS-sgRNA1 / NRPS-sgRNA2, FAS-sgRNA1 / FAS-sgRNA2, PKS1-sgRNA1 / PKS1-sgRNA2, and PKS2-sgRNA1 / PKS2-sgRNA2 used for each gene knockout.

[0091] The PCR reaction system was 10 μL of 5×FastPfu buffer, 1 μL of 10 mM dNTPs, 2.5 μL of upstream / downstream primers, 0.5 μL of DNA template, 1.5 μL of FastPfu (TransGene), and 32 μL of ultrapure water.

[0092] The PCR reaction conditions were: 95°C pre-denaturation for 5 min; 95°C denaturation for 30 sec, 55°C annealing for 30 sec, 72°C extension for 45 sec, 35 cycles; and finally 72°C extension for 10 min. The PCR product was purified and then subjected to PEG-mediated protoplast transformation.

[0093] (2) Preparation of donor DNA fragments

[0094] The preparation of donor DNA fragments was completed by a one-step PCR method. Upstream and downstream primers containing four gene homology arms were designed, and plasmid pSilent-1 (see Nakayashiki H, Hanada S, Quoc NB, Kadotani N, Tosa Y, Mayama S. RNA silencing as a tool for exploring gene function in ascomycetefungi. Fungal Genet Biol. 2005; 42: 275–83) was used as a template for PCR amplification to obtain donor DNA fragments for four gene knockouts, which were named MH-NRPS-sgRNA1 / NRPS-sgRNA2, MH-FAS-sgRNA1 / FAS-sgRNA2, MH-PKS1-sgRNA1 / PKS1-sgRNA2, and MH-PKS2-sgRNA1 / PK S2-sgRNA2. The PCR reaction system and reaction conditions were the same as above, and the relevant primer sequences are shown in Table 2.

[0095] Table 2 Primers used for donor DNA preparation

[0096]

[0097] (3) PEG-mediated protoplast transformation

[0098] The preparation of Aspergillus niger M202 protoplast suspension refers to patent CN112250740A. Take 100μL protoplast suspension, 2μg pCas9, 2μg sgRNA expression cassette fragment and 2μg donor DNA, add 1mL solution C (Tris-HCl10mM, CaCl25.54g / L, PEG6000 50% (w / v), pH 7.5) ice bath for 10min, 2mL solution B and mix. After mixing with the preheated upper culture medium MMSH containing hygromycin, spread it on the lower culture medium MMSH plate. Incubate the plate in a 30℃ incubator for 3-5 days until transformants grow.

[0099] (4) Genotype verification of gene-inactivated strains

[0100] The genomic DNA of the transformants after secondary subculture and purification was extracted using the Tiangen new plant genome extraction kit DP350. Then, the extracted genomic DNA was used as a template to perform gene PCR verification on the transformants using the genomic upper and lower verification primers of each gene. The specific sequences of the primers are shown in Table 3.

[0101] Table 3 Verification primers for each gene inactivated strain

[0102]

[0103] The PCR reaction system and PCR reaction conditions were the same as above. The PCR amplification products were subjected to 1% agarose gel electrophoresis (150V voltage, 20 minutes), the gene amplification bands were observed under the gel imaging system, and each PCR product was sequenced and verified, and finally four strains with knockout genes encoding predicted proteins were obtained, which were named AnSM1, AnSM2, AnSM3 and AnSM4.

[0104] Example 3. Citric acid fermentation of gene-inactivated strains

[0105] The gene-inactivated strains AnSM1, AnSM2, AnSM3 and AnSM4 and the control strain M202 were inoculated on PDA medium and cultured at 30°C for 5 days. Then, spores were collected with 0.9% saline and counted using a hemocytometer. 6 The inoculum size of 1 mL was inoculated into citric acid fermentation medium (corn starch medium, total sugar content 12%) and cultured at 34°C and 250 r / min for 96 h.

[0106] The fermentation supernatant was collected by rapid filtration, diluted 10 times, heated and boiled for 10 minutes, filtered through a filter membrane and tested for citric acid content by HPLC. The specific detection conditions refer to patent CN112250740A. The results showed that the inactivation of each gene can significantly promote the production of citric acid. Compared with the control strain, the citric acid fermentation levels of AnSM1, AnSM2, AnSM3 and AnSM4 increased by 15-25%, 20-30%, 15-30% and 15-25% respectively.

[0107] Example 4 Citric acid fermentation of a strain with inactivated whole gene cluster

[0108] Further analysis of the above four genes revealed that they were all located in a gene cluster of unknown function. An09g01690 was the first gene in this gene cluster, and An09g01930 was the last gene in this gene cluster. The entire gene cluster was 67,885 bp in length and included 24 genes. Its nucleotide sequence is shown in SEQ ID NO:5.

[0109] The method of Example 1-2 was used to construct a strain with the entire gene cluster inactivated. Among them, the pNRPS-sgRNA1 and pPKS2-sgRNA2 expression plasmids constructed in Example 1 were used as templates, and 5S-Fm and sgRNA-Rm in Example 2 were used as primers to amplify the sgRNA expression cassette fragment NRPS-sgRNA1 / PKS2-sgRNA2 of the entire gene cluster; MH-NRPS-sgRNA1-F and MH-PKS2-sgRNA2-R were used as primers, and plasmid pSilent-1 was used as a template to amplify the donor DNA fragment MH-NRPS-sgRNA1 / PKS2-sgRNA2 of the entire gene cluster; 2 μg of pCas9, 2 μg of sgRNA expression cassette fragments and 2 μg of donor DNA were transformed into Aspergillus niger M202 using the method of Example 2, and NRPS-gF and PKS2-gR were used as primers to perform PCR verification on the transformants, and finally the engineered strain AnSMc with the entire gene cluster inactivated was obtained. The method of Example 3 was used to perform fermentation verification on the engineered strain AnSMc and the control strain M202. The results showed that the inactivation of the entire gene cluster could also significantly increase the yield of citric acid by 15-30%.

[0110] Example 5 Gene (cluster) inactivation and citric acid efflux enhancement to improve citric acid fermentation level

[0111] The overexpression plasmid pGm-CexA (see patent CN112250740A) of citric acid efflux protein was transformed into Aspergillus niger strain M202 using the method of Example 2 to obtain the overexpression strain AnCex of citric acid efflux protein. Then, the methods of Examples 1-4 were used to knock out the four genes and the entire gene cluster in the citric acid high-yield strain AnCex, respectively, to obtain inactivated strains AnCSM1, AnCSM2, AnCSM3, AnCSM4 and AnCSMc. The above strains were fermented and verified using the method in Example 3, while the wild-type strain M202 was used as a control. The results showed that the citric acid fermentation levels of AnCSM1, AnCSM2, AnCSM3, AnCSM4 and AnCSMc could be increased to 2.01-2.30 times that of the control strain, indicating that on the basis of citric acid efflux enhancement, the inactivation of the unknown function gene cluster or the related proteins therein can further improve the production of citric acid. Sequence Listing <110> Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences <120> Microorganism producing citric acid and its construction method and application <160> 40 <170> SIPOSequenceListing 1.0 <210> 1 <211> 2793 <212> PRT <213> Aspergillus niger <400> 1 Met Ala Pro Ile Ala Leu Ser Thr Pro Gln Pro Ser His Leu Ser Ile 1 5 10 15 Pro Ser Lys Ser Phe Lys Ser Ile Lys Asp Asp Thr Thr Ile Val Thr 20 25 30 Arg Glu Pro Ala Ser Ile Asp Gly Phe Leu Pro Val Glu Val Lys Thr 35 40 45 Gly Asp Thr Ala Glu Val Ser Gly Val Leu Cys Thr Ala Phe Gly Leu 50 55 60 Ala Leu Arg Cys Tyr Asn Thr Glu Gly Val His Phe Glu Tyr His His 65 70 75 80 Glu Thr Ala Thr Gly Ser Ser Ser Thr Asn Leu Leu Phe Glu Leu Asp 85 90 95 Glu Phe Arg Lys Val Glu Asp Leu Leu Gln Ala Val Lys Ala Thr Leu 100 105 110 His Ser Ala Arg Ser Ala Leu Glu Thr Pro Asp Glu Lys Gln Gln Thr 115 120 125 Gln Ser Tyr Gln Ser Phe Arg Ser Ala Val Ile Phe Arg Glu Glu Ser 130 135 140 Leu Asn Ser Lys His Ile Ser Pro Ser Pro Asn Phe Asp Leu Glu Leu 145 150 155 160 His Ala Thr Asn Ser Arg Asn Gly Ile Glu Ile His Met Leu Tyr Thr 165 170 175 Pro Ser Thr Val Ser Glu Asp Leu Ala Val His Leu Ala Asp Thr Leu 180 185 190 His Arg Thr Phe Glu Leu Val Asn Asn His Ser Glu Thr Glu Leu Ser 195 200 205 Ser Ala Ser Tyr Leu Gly Pro Leu Ser His Lys Arg Ile Phe Asn Trp 210 215 220 Asn Ser Asn Ala Pro Asn Ala Val Ile Glu Cys Ala His Glu Leu Val 225 230 235 240 Arg Lys Thr Ala Ser Leu Gln Pro Gln Ala Pro Ala Ile Asp Ser Trp 245 250 255 Asp Ala Gln Met Thr Tyr Ala Glu Leu Asp Ser Leu Ser Asn His Leu 260 265 270 Ala Thr Val Leu Val Glu Leu Gly Ile Lys Ala Glu Val Ile Val Pro 275 280 285 Leu Cys Phe Glu Lys Ser Ala Trp Tyr Val Val Ala Leu Leu Ala Val 290 295 300 Leu Lys Ala Gly Gly Ala Phe Val Pro Leu Asp Pro Thr His Pro Ser 305 310 315 320 Ser Arg Leu Arg Glu Ile Val Ser Gln Val Asp Ala Pro Ile Ile Leu 325 330 335 Thr Thr Ala Lys His His Asp Leu Phe Asp Asp Phe Ser Val Lys Thr 340 345 350 Met Ile Val Asn Glu Asp Thr Val Gly Thr Ile Thr Ser Gly Asp Gly 355 360 365 Cys Leu Thr Pro Val Thr Val Thr Pro Glu Asn Leu Ala Tyr Val Ile 370 375 380 Phe Thr Ser Gly Ser Thr Gly Lys Pro Lys Gly Thr Met Ile Gln His 385 390 395 400 Gln Ser Phe Cys Ser Gly Ser Leu Arg Gln Gly Glu Ala Ala Asn Met 405 410 415 Gly Pro His Ser Arg Ile Phe Gln Phe Ala Ser Tyr Ser Phe Asp Val 420 425 430 Ser Ile Leu Glu Ile Leu Thr Gly Leu Ile Phe Gly Ala Cys Ile Cys 435 440 445 Ile Pro Asn Glu Gln Leu Gly Arg Ala Asp Phe Val Gln Ser Met Asn 450 455 460 Asp Phe Arg Val Asn Trp Ala Phe Leu Thr Pro Ser Val Leu Lys Val 465 470 475 480 Leu Thr Pro Asp Gln Leu Pro Leu Leu Lys Thr Leu Val Leu Gly Gly 485 490 495 Glu Ala Met Ser Glu Ser Asp Met Met Thr Trp Ala Gly Arg Leu Gln 500 505 510 Leu Met Asn Gly Tyr Gly Pro Ser Glu Cys Ser Val Ala Ala Thr Ala 515 520 525 Asn Thr Gln Leu Asn Pro Gln Ser Ser Pro Arg Asn Ile Gly Arg Ala 530 535 540 Ile Gly Gly Val Cys Trp Ile Val Asp Pro Gln Asn His Asn Val Leu 545 550 555 560 Leu Pro Val Gly Ala Thr Gly Glu Leu Ile Ile Gln Gly Pro Ile Val 565 570 575 Ser Arg Gly Tyr Ile Lys Asp Pro Val Lys Thr Ser Ser Ala Phe Phe 580 585 590 Arg Asn Pro Ala Trp Met Val Pro Gly Thr Ser Ser Ile Trp Asn Arg 595 600 605 Phe Tyr Lys Thr Gly Asp Leu Val Arg Tyr Asp Ala Asp Gly Met Ile 610 615 620 His Phe Ile Gly Arg Lys Asp His Gln Val Lys Leu Asn Gly Gln Arg 625 630 635 640 Met Glu Leu Gly Glu Val Glu His His Leu Trp Thr Asp Pro Val Ile 645 650 655 Arg Asn Gly Met Ala Leu Ile Pro Tyr Thr Gly His Cys Lys Ser Arg 660 665 670 Leu Val Gly Ile Val Ser Leu His Lys His Phe Thr Thr Ala Ser Ile 675 680 685 Gln Thr His Thr Ser Ile Arg Leu Val Asn His Ala Glu Ala Ala Ala 690 695 700 Asp Thr Gln Ala Ile Ala Glu Arg Leu Ser His Leu Leu Pro Thr Tyr 705 710 715 720 Met Val Pro Thr Ile Trp Ile Val Leu Glu Ser Leu Pro Leu Met Ser 725 730 735 Ser Gly Lys Met Asp Arg Lys Lys Val Gly Asp Phe Val Ala Asp Met 740 745 750 Ser Thr Glu Ala Phe Gln His Ile Ile Gly Asn Ser Thr Lys Pro Gly 755 760 765 Asp Lys Ala Ser Trp Thr Lys Thr Glu Ile Leu Val Gln Lys Val Trp 770 775 780 Ala Lys Ala Leu Asn Val Pro Glu Glu Gln Ile Gly Lys His Ser Ser 785 790 795 800 Phe Leu Ser Val Gly Gly Asp Ser Ile Lys Ala Met Ala Val Met Ser 805 810 815 Glu Phe Arg Lys Leu Gly Val Thr Val Leu Val Ala Asp Leu Leu Arg 820 825 830 Tyr Asn Val Thr Glu Ile Ala Ser Lys Leu Glu Glu Ser Gly Ser Ala 835 840 845 Asp Val Leu Arg Arg Ser Glu Thr Asp His Leu Pro Ile Ser His Leu 850 855 860 Gln Arg Thr Phe Leu Glu Ala Ser Arg Ser Gly Asn Ser Ala Ile His 865 870 875 880 His Ala Met Leu Leu Glu Leu Asn His Lys Ile Ser Ile Gln Gln Leu 885 890 895 Asn Asp Ala Leu Asp Ser Ile Val Ser Ser Tyr Pro Ala Leu Thr Leu 900 905 910 Gly Phe Asn Glu Gln Asn Gly Thr Trp Leu Gln Thr Phe Arg Asp His 915 920 925 Asp Glu Leu Glu Asn Pro Val Tyr His Leu Asn Leu Asn His Val Ser 930 935 940 Ser Leu Glu Glu Ala Gln Glu Ile Met Gln His Arg Gln Ala Ser Val 945 950 955 960 Gly Val Ser Gly Asn Pro Asn Leu Met Ile Asp Leu Phe Ser Leu Arg 965 970 975 Gly Thr Gln His Ile Ser Leu Val Val His Ala Ala Ile Leu Asn Ser 980 985 990 Gln Ser Leu Ser Thr Val Ile Gly His Leu Asp Tyr Leu Leu Gln Ser 995 1000 1005 Asn Asp Glu Lys Glu Glu Asn Val Trp Glu Ser Lys Leu Leu Ser Arg 1010 1015 1020 Asp Glu Phe Ser His Ala Trp Leu Asp His Leu Arg Ser Asp Thr Gly 1025 1030 1035 1040 Met Glu Pro Met Asn Gly Pro Leu Gln Ser Val Ala Leu Glu Ser Leu 1045 1050 1055 Ser Gly Tyr Arg Asp Tyr Val Thr Glu Thr Leu Glu Leu Gly Ser Glu 1060 1065 1070 Ile Thr Gly Leu Leu Thr Gly Pro Cys Asn Leu Lys Phe Gly Thr Asn 1075 1080 1085 Ala Gly Asp Ile Phe Leu Ala Thr Leu Ile Leu Ser Tyr His Gln Ile 1090 1095 1100 Leu Glu Glu Asn Asp His Ser Thr Leu Ala Ile Val Glu Thr Thr Ala 1105 1110 1115 1120 Thr Glu Ser Thr Leu Ser Lys His Leu Arg Gly His Val Gly Gln Phe 1125 1130 1135 Ser Ser Leu Gln Pro Val His Val Pro Val Glu His Ser Asp Gln Val 1140 1145 1150 Thr Leu Gln Leu Ile Arg Asp Thr Lys Asp Lys Val Arg Gln Arg Gly 1155 1160 1165 Ile Gly Ser Lys Leu Gly Ser Ser Asn Val Ile Phe Asn Tyr Gly Leu 1170 1175 1180 Ser Ser Arg Leu Thr Glu Gly Ile Val Leu Lys Gln Ala Thr Leu Thr 1185 1190 1195 1200 Arg Arg His Gly Leu Gly Ala Ile Val Pro Thr Thr Phe Glu Val Ser 1205 1210 1215 Cys Leu Leu Glu Asp Gly Gln Phe Arg Ile Thr Val Gly Ser Asp Gln 1220 1225 1230 Val Ser Ala Gln Lys Leu Leu Leu Phe Thr Asp Gln Tyr Arg Gln Asn 1235 1240 1245 Ile Thr Gln Ala Val Glu Ile Leu Ala Ser Asn Thr Ala Lys Glu Phe 1250 1255 1260 Thr Leu Ala Asp Phe Pro Leu Leu Lys Leu Asp His Pro Ser Leu Glu 1265 1270 1275 1280 Arg Val Asn Arg Gln His Leu Pro Arg Val Gly Leu Arg Pro Asn Gln 1285 1290 1295 Ile Glu Asp Met Tyr Pro Cys Ser Pro Leu Gln Gln Gly Leu Leu Met 1300 1305 1310 Ser Gln Ala Arg Lys Gln Gly Ser Tyr Asn Val Phe Ile Gly Trp Glu 1315 1320 1325 Val Val Ser Cys Gly Ala Asn Asp Thr Val Asp Ile Gly Arg Leu Leu 1330 1335 1340 Ser Ser Trp Gln Lys Val Val Asp His His Pro Ala Leu Arg Thr Ile 1345 1350 1355 1360 Phe Thr Asp Glu Phe Gly Gly Lys Glu Pro Tyr Ala Gln Ile Val Leu 1365 1370 1375 Arg Arg Pro Arg Ala Glu Val Ser Val Lys Pro Cys Ser Asp Val Asp 1380 1385 1390 Val Ala Ser Val Leu Glu Glu Thr Ile Glu Leu Gly Thr Asn Pro Val 1395 1400 1405 Val Pro Pro His Arg Phe Ser Ile Asn Val Thr Ala Asp Asp Arg Val 1410 1415 1420 Phe Cys Arg Val Asp Ala Ser His Thr Ile Val Asp Gly Val Ser Lys 1425 1430 1435 1440 Ser Leu Ile Leu Arg Asp Leu Lys Arg Ala Tyr Ala Gly Thr Leu Leu 1445 1450 1455 Ser Pro Thr Gly Gly Ala Lys Tyr Arg Asp Phe Val Gln Tyr Val Ala 1460 1465 1470 Ala Asn Asp Met Ser Ala Ser Ile Ser Phe Trp Gln Asn His Leu Ala 1475 1480 1485 Glu Val Glu Pro Cys Tyr Phe Pro Lys Leu Ala Glu Ala Glu Gly Thr 1490 1495 1500 Pro Glu Leu Lys Tyr Val Gln Val Glu Leu Gly Ile Pro Thr Ser Val 1505 1510 1515 1520 Leu Arg Asp Phe Cys Val Ser Gln Ser Ile Thr Ile Ser Asn Leu Leu 1525 1530 1535 Gln Thr Val Trp Ala Leu Val Cys Arg Val Tyr Val Gly Arg Asp Asp 1540 1545 1550 Ile Cys Phe Gly Tyr Leu Ala Ser Gly Arg Asp Ala Pro Val Glu Asp 1555 1560 1565 Leu Val Asn Met Val Gly Pro Thr Ile Ser Val Leu Ile Cys Lys Ala 1570 1575 1580 Leu Leu Glu Asp Ser Thr Ile Ile Lys Asp Ala Leu Lys Ala Cys Gln 1585 1590 1595 1600 Glu Glu Phe Leu Asp Ser Leu Ala His Gln His Cys Ser Leu Ala Glu 1605 1610 1615 Ile Gln His Ser Leu Gly Leu Ala Gly Met Pro Leu Phe Asn Thr Gly 1620 1625 1630 Ile Ser Phe Gln Thr Met Thr Asp Val Asp Ser Asp Asp Val Ile Thr 1635 1640 1645 Asn Ile Asp Phe Glu Gly Leu Ile Val Arg Glu Pro Thr Glu Tyr Asn 1650 1655 1660 Ile Thr Val His Val Val Asp Ser Pro Asn Asn Val Asp Ile Lys Leu 1665 1670 1675 1680 Gly Tyr Trp Thr Asp Cys Ile Ser Glu Ser Arg Ala Gly Ser Val Ser 1685 1690 1695 Gly Thr Phe Ser Ala Ile Leu Lys Ser Val Val Glu His Pro Glu Ala 1700 1705 1710 Arg Val Asp Gln Leu Asn Val Ile Gly Arg Tyr Asp Leu Ser Gln Ile 1715 1720 1725 Val Ser Trp Asn Gln Asp Val Pro Glu Ile Ser Ser Tyr Thr Val Pro 1730 1735 1740 Asp Leu Val Leu Gln Gln Ile Thr Arg Thr Pro His Ala Ile Ala Ile 1745 1750 1755 1760 Asp Thr Met Ala Glu Lys Ile Thr Tyr Gln Asp Phe Gly Arg Met Trp 1765 1770 1775 Leu Ser Leu Ser Arg Tyr Leu Thr Arg Leu Gly Ile Gly Ala Gly Asp 1780 1785 1790 Tyr Val Pro Leu Thr Phe Glu Lys Ser Ala Trp Ala Ile Val Ala Met 1795 1800 1805 Phe Ala Val Leu Gly Thr Gly Ala Ala Phe Val Pro Leu Asp Pro Lys 1810 1815 1820 Thr Pro Val Glu Arg Leu Arg Glu Ala Ser Ile Gln Thr Ala Ala Asn 1825 1830 1835 1840 Val Val Leu Ala Ser Pro Lys Tyr Ala Ser Asn Trp Asn Asp Leu Ile 1845 1850 1855 Pro Thr Ile Val Ser Val Asp Gln Asp Leu Leu Asp Lys Leu Asn Glu 1860 1865 1870 Glu Gln Asp Asp Leu Asp Gln Gly Val Val Arg Ala Arg Pro His His 1875 1880 1885 Asp Ala Tyr Val Ile Phe Thr Ser Gly Ser Thr Gly Lys Pro Lys Gly 1890 1895 1900 Cys Val Val Gln His Ser Ala Phe Cys Ser Gly Ala Leu Ala Gln Gly 1905 1910 1915 1920 Lys Leu Ala Cys Leu Gly Pro Ser Ser Arg Val Leu Gln Phe Ala Ser 1925 1930 1935 Tyr Ser Phe Asp Val Ser Leu Leu Glu Ile Met Thr Ser Leu Met Phe 1940 1945 1950 Gly Ala Cys Ile Cys Val Pro Asp Glu Asp Leu Ser Lys Asp Ile Lys 1955 1960 1965 His Cys Ile Asn Val Leu Asn Ile Asn Trp Thr Phe Leu Thr Pro Ser 1970 1975 1980 Val Leu Lys Leu Leu Gln Pro Ser Asp Val Pro Ser Leu Lys Thr Leu 1985 1990 1995 2000 Val Leu Gly Gly Glu Ala Leu Ser Lys Gly Asp Ile Leu Asn Trp Ala 2005 2010 2015 Asp Arg Val Gln Leu Tyr Asn Gly Tyr Gly Pro Ser Glu Cys Ser Val 2020 2025 2030 Ala Ala Ala Ala Asn Pro Gly Leu Gln Pro Thr Thr Asp Pro Ala Asn 2035 2040 2045 Ile Gly His Ala Val Gly Gly Val Leu Trp Ile Val Asp Ala Lys Gln 2050 2055 2060 Pro Ser Lys Leu Leu Pro Ile Gly Ala Val Gly Glu Leu Leu Ile Ser 2065 2070 2075 2080 Gly Pro Ile Leu Ala Arg Gly Tyr Leu Gly Asp Pro Glu Lys Thr Ala 2085 2090 2095 Ala Ala Phe Val Lys Gln Lys Ser Phe Val Pro Gly Ser His Lys Arg 2100 2105 2110 Glu Arg Tyr Tyr Arg Thr Gly Asp Leu Val Arg Tyr Asn Ala Asp Gly 2115 2120 2125 Thr Ile His Phe Ile Gly Arg Ala Asp Gly Gln Val Lys Ile Arg Gly 2130 2135 2140 Gln Arg Val Glu Leu Gly Glu Ile Glu Tyr Asn Ile Glu Arg Asp Glu 2145 2150 2155 2160 Asn Ile His Gln Ala Leu Thr Leu Leu Pro Ser Gln Gly Pro Phe Lys 2165 2170 2175 Lys Gln Leu Leu Ala Val Val Ser Phe Lys Gly Phe Asp Leu Pro Val 2180 2185 2190 Glu Glu Gly Arg Leu Thr Leu Val Pro Glu Asp Arg Arg Pro Lys Ile 2195 2200 2205 Met Ala Val Val Thr Arg Ile Ala Asn Thr Ile Ser Ala Ala Leu Pro 2210 2215 2220 Ile Tyr Met Val Pro Ala Leu Trp Val Pro Val Asn Ala Ile Pro Leu 2225 2230 2235 2240 Leu Pro Ser Gly Lys Leu Asp Arg Lys Lys Val Arg Arg Trp Val Glu 2245 2250 2255 Thr Leu Asp Asp Ala Thr Tyr Glu Gln Ile Ala Asn Val Gly Ser Lys 2260 2265 2270 Ala Ser Gly Arg Gly Pro Ser Thr Pro Ala Glu Phe Glu Leu Arg Arg 2275 2280 2285 Ile Trp Ala Ser Val Leu Asn Arg Pro Glu Glu Gln Ile Ser Met Glu 2290 2295 2300 Arg Ser Phe Gln Ser Leu Gly Gly Asp Ser Ile Ser Ala Met Gln Val 2305 2310 2315 2320 Ile Ala Gln Cys Arg Asp Ala Lys Leu Ser Leu Ala Val Lys Asp Leu 2325 2330 2335 Ile Gln Cys Pro Thr Ile Ala Glu Leu Ala Pro Arg Val Arg Pro Leu 2340 2345 2350 Asp Glu Asp Ala Glu Pro Met Glu Asp Asp Ile Asp Val Pro Phe Asn 2355 2360 2365 Leu Ser Pro Gln Gln Thr Trp Glu Phe Asp Leu Lys Gln His Cys Asp 2370 2375 2380 Leu Asn Arg Tyr Asn Thr Thr Phe Leu Leu Ser Val His Ser Asp Ser 2385 2390 2395 2400 Asp Leu Lys Pro Val Val Asp Ala Thr Val Glu Arg His Pro Met Leu 2405 2410 2415 Arg Ala Arg Phe Gln Arg Asp Glu Ser Gly Arg Trp Met Gln Tyr Ile 2420 2425 2430 Pro Leu Asp Arg Ser Asn Ser His Leu Tyr Gln Glu Leu Glu Val Pro 2435 2440 2445 Asp Leu Glu Ala Ile Glu Pro Tyr Thr Ala Pro Asn Gln Ser Gly Phe 2450 2455 2460 Asp Leu Gln Arg Gly Pro Leu Met Arg Ala Ile Ile Phe Lys Leu Pro 2465 2470 2475 2480 Asn Gly Asp Arg Tyr Val Gly Leu Thr Val His His Met Val Ile Asp 2485 2490 2495 Thr Val Ser Trp Arg Ile Leu Leu Gln Asp Met Gln Gln Gly Leu Ser 2500 2505 2510 Thr Gly Thr Ile Thr Ser His Arg Ser Asp Ser Phe Ala Leu Trp Cys 2515 2520 2525 Arg Lys Leu Lys Glu His Ala Asp Glu Lys Trp Thr Pro Glu Tyr Val 2530 2535 2540 Leu Pro Phe Ser Ile Pro Ala Pro Leu Tyr Asp Tyr Trp Gln Met Glu 2545 2550 2555 2560 Glu Ser Met Asn Leu Phe Val Thr Glu Thr Ile Gln Glu Phe Ser Leu 2565 2570 2575 Asp Thr Lys Thr Thr Ala Arg Leu Met Gly Asn Cys Asn His Gln Tyr 2580 2585 2590 Gln Thr Lys Pro Leu Asp Ile Phe Leu Ser Ala Ile Phe His Ala Phe 2595 2600 2605 Ser Ala Thr Phe Thr Asp Arg Pro Val Pro Ala Val Phe Ile Tyr Ser 2610 2615 2620 His Gly Arg Asp Glu Phe Gly Gly Asp Leu Asp Val Ser Asn Thr Ile 2625 2630 2635 2640 Gly Trp Phe Thr Ser Ser Thr Pro Val Val Ile Asn Pro Ser Gly Asp 2645 2650 2655 Ile Val Asp Asp Leu Gln Ala Val Arg Asp Ala Arg Ala Ser Val Pro 2660 2665 2670 Gly Asn Gly Val Pro Tyr Trp Ala Thr Arg Trp Leu Thr Asp Asp Gly 2675 2680 2685 His Ala Ala Phe Glu His His Ser Pro Phe Glu Leu Ser Met Asn Tyr 2690 2695 2700 Leu Gly Gln Tyr Gln Gln Leu Glu Arg Asp Asp Ser Val Leu Arg Tyr 2705 2710 2715 2720 Val Asp Val Lys Lys Pro Lys Ala Thr Gly Ser Gly Ser Gly Leu Leu 2725 2730 2735 Arg Thr Ala Leu Val Glu Thr Gln Ala Ile Val Ile Gly Asp Ser Leu 2740 2745 2750 Gln Ile Lys Phe Val Tyr Asn Thr Arg Met Ala Arg Gln Lys Gln Leu 2755 2760 2765 Leu Asp Trp Gln Ala Lys Val Lys Asp Ser Leu Leu Glu Ile Ala Asn 2770 2775 2780 Arg Leu Gly Asp Gly Val Glu Thr Glu 2785 2790 <210> 2 <211> 2108 <212> PRT <213> Aspergillus niger <400> 2 Met Met Thr Val Lys Thr Pro Leu Ser Ser Leu Ser Ser Glu Phe Ala 1 5 10 15 Gly Leu Thr Pro Glu Ile Ser Arg Thr Ser Ser Val Thr Gly Asp Lys 20 25 30 Arg Ser Leu Ile Ala Ala Tyr Gly Ser Val Asn Thr Thr Ile Ser Leu 35 40 45 Pro Thr Leu Gln Leu Pro Leu Ala Asn Gln Leu Trp Lys Asp Phe Arg 50 55 60 Ser Asn Leu Pro Ser Ser Ser Pro Glu Asn Asp Leu Thr Ile Lys Glu 65 70 75 80 Gly Leu Val Ala Glu Phe Ile Asn Phe Cys Ile Thr Arg Ser Ser Asp 85 90 95 Asp Thr His Cys Asp Asp Pro Ala Ala Val Leu Gln Phe Leu Glu Ser 100 105 110 Leu Leu Glu Ala Phe Glu Gln Asp Phe Val Gly Gly Asn Asp Val His 115 120 125 Ser Ala Val Ser His Leu Asp Thr Gln His Lys Thr Pro Ile Ile Lys 130 135 140 Ala Tyr Phe Ala Ala Tyr Gly Thr Leu Ser Arg Lys Pro Arg Arg Gln 145 150 155 160 Pro Ser Ile Leu Leu Glu Ser Val Thr Asp Gly Pro Ser Arg Ile Tyr 165 170 175 Ala Ile Phe Gly Gly Gln Gly Asn Asp Glu Lys Tyr Phe Ala Asp Leu 180 185 190 Lys Asp Thr Tyr Ala Thr Tyr Thr Asp Ile Leu Arg Arg Cys Val Phe 195 200 205 Ser Gly Ala Arg Leu Phe Glu Asp Leu Ala Gly Ala Ser Val Phe Lys 210 215 220 Thr His Phe Gln His Gly Phe Ala Leu Met Asp Trp Leu Glu Lys Pro 225 230 235 240 Glu Ser Glu Pro Glu Thr Ser Tyr Leu Leu Ser Ala Pro Val Ser Met 245 250 255 Pro Leu Ile Gly Leu Leu Gln Met Cys His Phe Ile Val Ala Cys Gln 260 265 270 Thr Met Gly Leu Ser Pro Arg Glu Met His Gly Tyr Leu Ser Gly Val 275 280 285 Thr Gly His Ser Gln Gly Ile Val Val Ala Leu Val Val Ser Val Ser 290 295 300 Asp Thr Trp Glu Asp Phe Tyr Arg His Ser Leu Glu Ala Leu Arg Ile 305 310 315 320 Leu Phe Phe Ile Ser Cys Arg Ser His Glu Ile Phe Pro Pro Gln Phe 325 330 335 Ile Pro Glu Ser Ile Arg Gln Asp Ala Glu Asp Asn Gly Glu Gly Thr 340 345 350 Pro Gly Pro Met Leu Arg Val Arg Asn Leu Arg Gln Gly Gln Leu Gln 355 360 365 Lys Ile Val Asp Glu Val Asn Ser Tyr Leu Pro Asp Glu Ser His Val 370 375 380 Ala Ile Ser Leu Phe Asn Gly Pro Gln Asn Leu Val Val Thr Gly Ser 385 390 395 400 Pro Leu Ser Leu Tyr Gly Leu Ser Arg His Leu Arg Gly Leu Lys Pro 405 410 415 Pro Asn Thr Ser Arg Glu Ala Gln Ile Pro Ala Ser Lys Arg Lys Pro 420 425 430 Lys Ile Glu His Arg Phe Leu Pro Ile Thr Ala Pro Phe His Ser Ile 435 440 445 Tyr Leu Asp Arg Val Val Ser Leu Val Met Lys Asp Leu Gly Asp Ile 450 455 460 Asn Ile Thr Gly Asn Gln Leu Arg Ile Pro Leu Tyr Ala Thr Thr Asp 465 470 475 480 Gly His Asp Met Arg Ser His Gly Ala Ser Asn Leu Ile Gly Asp Ile 485 490 495 Val Ser Met Ile Ala Arg Glu Pro Val His Trp Glu Asn Ala Thr Ile 500 505 510 Leu Pro Asn Ala Thr His Ile Ile Asp Phe Gly Pro Gly Gly Ser Ala 515 520 525 Gly Val Gly Ala Val Thr Met His Asn Lys Ala Gly Gln Gly Val Arg 530 535 540 Leu Ile Leu Gly Gly Ser Met Lys His Ser Glu Glu Tyr Gly Asn Lys 545 550 555 560 Ala Glu Leu Phe Asn Arg Asp Cys Gly Tyr Pro Val Val Tyr Ala Glu 565 570 575 Asn Trp Ala Glu Val Tyr Arg Pro Arg Leu Val Arg Asp Ala Ala Gly 580 585 590 Lys Val Ile Val Gln Thr Lys Phe Ser Glu Leu Leu Gly Leu Pro Pro 595 600 605 Ile Met Val Ala Gly Met Thr Pro Thr Thr Val Pro Trp Asp Phe Val 610 615 620 Ser Ala Thr Met Asn Ala Gly Tyr His Ile Glu Leu Ala Gly Gly Gly 625 630 635 640 Tyr Tyr Ser Lys Gly Ala Leu Glu Glu Ala Ile Asn Lys Ile Val Gln 645 650 655 Asn Gly Ile Pro Gly Arg Gly Val Thr Cys Asn Ile Ile Tyr Ala Ser 660 665 670 Pro Asn Ser Val Arg Trp Gln Ile Pro Leu Leu Gln Glu Leu Arg Ala 675 680 685 Lys Gly Val Pro Ile Asp Gly Ile Thr Ile Gly Ala Gly Val Pro Ser 690 695 700 Val Asp Val Ala Asn Glu Tyr Val Glu Thr Leu Gly Leu Lys His Ile 705 710 715 720 Ala Phe Lys Pro Gly Ser Val Glu Ala Ile Gln Gln Val Ile Asp Ile 725 730 735 Ala Arg Glu Asn Lys Ser Phe Pro Ile Ile Leu Gln Trp Thr Gly Gly 740 745 750 Arg Gly Gly Gly His His Ser Tyr Glu Asp Phe His Ala Pro Ile Leu 755 760 765 Glu Met Tyr Gly Arg Ile Arg Ala Cys Lys Asn Ile Val Leu Val Ala 770 775 780 Gly Ser Gly Phe Gly Gly Ala Asp Asp Thr Leu Pro Tyr Leu Thr Gly 785 790 795 800 Glu Trp Ser Lys Ala Phe Asn Ala Pro Ser Leu Met Pro Phe Asp Gly 805 810 815 Ile Leu Leu Gly Ser Arg Val Met Thr Ala Lys Glu Ala His Thr Ser 820 825 830 Ile Glu Ala Lys Gln Leu Ile Val Asp Ile Pro Gly Ile Asp Asp Ala 835 840 845 Val Trp Asp Gln Thr Tyr Thr Arg Pro Thr Gly Gly Ile Ile Thr Val 850 855 860 Arg Ser Glu Met Gly Glu Pro Ile His Lys Ile Ala Thr Arg Gly Val 865 870 875 880 Leu Phe Trp Ala Glu Met Asp Arg Thr Val Phe Ser Ile Ala Asp Lys 885 890 895 Lys Ala Arg Ala Ala Phe Leu Gln Glu Asn Lys Lys Arg Ile Ile Asp 900 905 910 Arg Leu Asn Asn Asp Phe Gln Lys Val Trp Phe Gly Arg Asp Ser Asp 915 920 925 Gly Lys Ala Cys Glu Leu Asn Glu Met Ser Leu Ser Ala Val Ala Trp 930 935 940 Arg Met Val Glu Leu Leu Tyr Val Lys His Gln Ala Arg Trp Ile His 945 950 955 960 His Ser Leu Gln Gln Leu Thr Phe Asp Phe Leu Gln Trp Leu Glu Asp 965 970 975 Val Val Gly Asn Ser Ser Gly Asn Phe Glu Ser Arg Tyr Trp Gln Thr 980 985 990 Val Gly Gln Ile Glu Glu Pro Phe Ser Ala Ile Glu Gln Phe Leu Ala 995 1000 1005 Lys His Ala Arg Ala Thr Ser Glu Leu Met Asp Val Pro Ser Thr Gln 1010 1015 1020 Met Phe Ile His Leu Cys Gln Arg Arg Gly Gln Lys Pro Val Pro Phe 1025 1030 1035 1040 Ile Pro Val Leu Asp Glu Asp Phe Glu Val Trp Phe Lys Lys Asp Ser 1045 1050 1055 Leu Trp Gln Ser Glu Asp Val Gln Ala Val Pro Gly Gln Asp Val Gly 1060 1065 1070 Arg Thr Cys Ile Leu His Gly Pro Val Ala Ala Lys His Thr Thr Val 1075 1080 1085 Pro Asn Glu Pro Ile Lys His Val Leu Asp Gly Ile His Ser Gln Trp 1090 1095 1100 Val Arg Ser Leu Thr Gly Leu Gly His Asp Asn Glu Gln Ala Gln Thr 1105 1110 1115 1120 Pro Leu Thr Glu Glu Gly Met Ser Glu Val Pro Ser Glu Ile Ser Leu 1125 1130 1135 Thr Arg Thr Tyr Asp Pro Met Leu Phe Gly Glu Glu Leu Pro Ser Ile 1140 1145 1150 Glu Pro Trp Met Gln Arg Leu Ala Gly Thr Gln Lys Gly Trp Arg His 1155 1160 1165 Ala Leu Phe Ser Gln Val Ser Ile Val Gln Gly Asn Ser Leu Cys Asp 1170 1175 1180 Asn Pro Ile Arg Arg Leu Phe Ala Pro Arg His Asn Thr Phe Val Glu 1185 1190 1195 1200 Val Glu Glu Cys Thr Ala Thr Glu Gln Ser Val Val Arg Leu Phe Glu 1205 1210 1215 Arg Thr Ala Asp Val Leu Gln Ile Leu Ala Ile Glu Val Arg Ala Val 1220 1225 1230 Ser Glu Asp Val Ile Lys Ala Thr Leu Phe Glu Asn Arg Thr Val Thr 1235 1240 1245 Gly Ala Pro Ala Gly Leu Glu Leu Leu Phe Thr Tyr His Pro Glu Phe 1250 1255 1260 Pro Phe Ala Pro Ile Arg Glu Val Ile Ser Asp Lys Ile Glu Arg Val 1265 1270 1275 1280 Lys Asp Phe Tyr His Gln Leu Trp Phe Gly Glu Pro Leu Ser Ser Ala 1285 1290 1295 Ser Val Leu Ser Gln Glu Asp Glu Phe Tyr Gly Asp Val Val Ala Val 1300 1305 1310 Thr Ala Glu Asp Ile Asn Lys Phe Asn Gln Cys Ile Arg Ser Thr Ala 1315 1320 1325 Thr Ser Arg Ser Arg Ser Ser Ser Gln Gln Thr Leu Asp Ala Pro Met 1330 1335 1340 Asp Tyr Ala Ile Val Val Ala Trp Lys Ser Val Met Lys Pro Leu Phe 1345 1350 1355 1360 Thr Lys Ser Ile Ser Gly Asp Leu Met Lys Leu Val His Leu Ser Asn 1365 1370 1375 Gln Phe Arg Arg Leu Gly Ser Phe Glu Pro Ile Lys Glu Gly Asp Val 1380 1385 1390 Leu Ser Ser Arg Ala His Ile Gln Ala Ile Leu Asn Glu Asp Ser Gly 1395 1400 1405 Lys Val Val Glu Val Ala Ala Leu Ile Ser Arg Asp Gly Val Asp Ile 1410 1415 1420 Val Glu Val Val Ser Arg Phe Met Tyr Arg Gly Ser Tyr Gly Asp Tyr 1425 1430 1435 1440 Asp Ala Ala Phe Gln Gln Thr Lys Glu Pro Lys Val Glu Leu Gln Leu 1445 1450 1455 Thr Ser Leu Lys Asp Val Val Val Leu His Ser Lys Pro Trp Phe Leu 1460 1465 1470 Pro Arg Asp Lys Asp Leu Asp Leu Lys Gly Tyr Lys Leu Thr Phe Glu 1475 1480 1485 Leu Glu Thr His Ala Gln Phe Gln Ser Lys Ser Val Phe Arg Gln Gln 1490 1495 1500 Arg Cys Phe Gly His Val Tyr Gly Arg Ser Pro Ala Gly Glu Thr Leu 1505 1510 1515 1520 Glu Val Ala Tyr Ile Asp His Thr Val Gly Val Ser Thr Ser Asn Pro 1525 1530 1535 Ile Met Asp Tyr Leu Lys Arg His Gly Ser Val Val Glu Gln Pro Val 1540 1545 1550 Leu Phe Gln Ser Pro Ile Thr Leu Gly Gly Arg Glu Ile Gln Phe Lys 1555 1560 1565 Ser Pro Ser Ser Asn Gln Asp Tyr Ala Ser Val Ser Gly Asp Tyr Asn 1570 1575 1580 Pro Ile His Val Ser Lys Thr Phe Ala Ala Tyr Ala Glu Leu Pro Asp 1585 1590 1595 1600 Thr Ile Thr His Gly Met His Met Ser Ala Arg Val Arg Gly Leu Leu 1605 1610 1615 Glu Gln Leu Thr Val Pro Gly Asn Pro Ser Gly Phe Lys Arg Tyr Gln 1620 1625 1630 Cys Asn Phe Val Gly Met Val Leu Pro Glu Asp Asp Val Glu Leu Val 1635 1640 1645 Leu Gln His Val Gly Met Val Asn Gly Arg Lys Leu Val His Ala Glu 1650 1655 1660 Ala His Arg Ala Gly Ser Gly Asp Lys Leu Ile Val Ala Glu Ala Glu 1665 1670 1675 1680 Ile Glu Gln Pro Ser Thr Gly Phe Leu Phe Thr Gly Gln Gly Ser Gln 1685 1690 1695 Glu Lys Gly Met Gly Met Glu Leu Tyr Ala Ser Ser Pro Ala Ala Arg 1700 1705 1710 Lys Val Trp Asp Leu Ala Asp Lys His Phe Leu Glu Asn Phe Gly Glu 1715 1720 1725 Tyr His Leu Phe His Met Ser Ala Phe Leu Thr Leu Pro Gly Phe Arg 1730 1735 1740 Ile Thr Asp Ile Val Arg Asn Asp Pro Arg Glu Leu Lys Ile His Phe 1745 1750 1755 1760 Gly Gly Ala Arg Gly Arg Ala Ile Arg Glu Asn Tyr Met Ala Leu Arg 1765 1770 1775 Cys Glu Met Ile His Ala Asp Gly Arg Thr Thr Ser Glu Lys Ile Phe 1780 1785 1790 Lys Asp Ile Thr His Asp Thr Lys Ser His Thr Phe Arg Ser Ser Thr 1795 1800 1805 Gly Leu Leu Ser Ser Thr Gln Phe Thr Gln Pro Ala Leu Thr Leu Met 1810 1815 1820 Glu Lys Ala Ile Ile Glu Asp Met Lys Asp Lys Gly Leu Leu Ala Asp 1825 1830 1835 1840 His Cys Ser Phe Ala Gly His Ser Leu Gly Glu Tyr Ser Ala Leu Ala 1845 1850 1855 Ala Ile Ala Glu Ile Met Pro Ile Glu Ser Leu Val Ser Val Val Phe 1860 1865 1870 Tyr Arg Gly Leu Thr Met Gln Met Ala Val Glu Arg Asp Glu Gln Gly 1875 1880 1885 Arg Ser Asn Phe Ser Met Cys Ala Val Asp Pro Ser Arg Leu Ser Lys 1890 1895 1900 Ser Phe Asn Glu Lys Arg Leu Gln Tyr Leu Val Ser Leu Ile Ala Arg 1905 1910 1915 1920 Glu Thr Ser Trp Leu Leu Glu Ile Val Asn Tyr Asn Val Ala Asn Ser 1925 1930 1935 Gln Tyr Val Cys Ala Gly Asp Leu Arg Ala Leu Asp Thr Leu Ser Ser 1940 1945 1950 Leu Ile Gly Glu Leu Lys Arg Arg Gln Ser Asp Phe Gln Ala Ile Leu 1955 1960 1965 Gln Asn Thr Gln Pro Glu Glu Glu Ile Arg Ala Ser Val Leu Asp Thr 1970 1975 1980 Leu His Gln Cys Ala Asn Ala Ala Lys Ala Lys Pro Gln Pro Ile Gln 1985 1990 1995 2000 Leu Glu Arg Gly Val Ala Thr Val Pro Leu Arg Gly Ile Asp Val Pro 2005 2010 2015 Phe His Ser Ser Phe Leu Ser Pro Gly Ile Thr Ala Phe Arg Ala Cys 2020 2025 2030 Leu Tyr Lys Tyr Ile Asp Lys Asp Thr Leu Asp Ala Asp Arg Leu Val 2035 2040 2045 Gly Lys Tyr Ile Pro Asn Leu Thr Ala Arg Pro Phe Glu Val Ser Glu 2050 2055 2060 Gly Tyr Phe Arg Gln Val Phe Arg Leu Thr Lys Ser Pro Ile Ile Ala 2065 2070 2075 2080 Lys Val Leu Ala Glu Trp Arg Glu Tyr Met Asp Gly Glu Arg Gly Leu 2085 2090 2095 Glu Lys Val Thr Arg Ser Ser Glu Val Val Val Ala 2100 2105 <210> 3 <211> 2604 <212> PRT <213> Aspergillus niger <400> 3 Met Val Thr Thr Thr Arg Ala Thr Asn Pro Thr Asn Thr Leu Leu Leu 1 5 10 15 Phe Gly Pro Gln Ala Leu Ser Phe Ser Thr Ala Thr Phe Ala Asp Ile 20 25 30 His Ala Arg Val Val Gln Thr Ser Glu Asn Ala Trp Ile Lys Gln Thr 35 40 45 Ile Thr Ser Leu Pro Gly Leu Trp Asp Ala Leu Val Lys Glu Phe Pro 50 55 60 Gln Tyr Gly Ala Leu Glu Gly Lys Gln Leu Leu Arg Asp Leu Asp Arg 65 70 75 80 Trp Phe Glu Thr Gly Thr Met Glu His Ala Glu Pro His Leu Pro Asn 85 90 95 Ile Leu Leu Ser Pro Met Val Val Ile Thr Gln Leu Thr Glu Tyr Val 100 105 110 Asp Tyr Leu Lys Thr Met Pro His Ala Ala Asp Gln Gln Thr Glu Thr 115 120 125 Val Gly Phe Cys Thr Gly Leu Leu Thr Ala Leu Ala Ala Ser Leu Ala 130 135 140 Pro Asp Ile Lys Gly Ile Arg Gln Tyr Gly Ala Ile Ala Ile Lys Leu 145 150 155 160 Ala Met Ile Ile Gly Gly Val Val Asp Val Gln Asp Ile Thr Ser Pro 165 170 175 Asn Gly Pro Ser Lys Ser Leu Ala Val Ala Trp Asp Ser Ala Glu Thr 180 185 190 Gln Asp Arg Leu Asn Gln Ile Ile Asp Gln Ser Pro Glu Ala Tyr Ile 195 200 205 Ser Val Glu Tyr Asp His Asn Arg Pro Thr Ile Thr Thr Ala Ala Arg 210 215 220 Ser Ile Ser Ser Leu Gln Gln Arg Leu Arg Asn Ala Gly Leu Ile Ala 225 230 235 240 Ser Gln Ile Gly Leu Arg Gly Arg Phe His Cys Ala Cys Tyr Glu Lys 245 250 255 Asp Ile Glu Ala Leu Ser Lys Phe Cys Asp Ser Val Pro Ser Leu Cys 260 265 270 Leu Pro Asp Ala Ala Val Leu Ala Leu Pro Thr Arg Ser Asn Asp Ala 275 280 285 Gly Ser Phe Ile Leu Ser Gly Lys Leu His His Cys Ala Leu Arg Ser 290 295 300 Ile Leu Leu Asp Thr Ser His Trp Tyr Gln Thr Leu Glu Val Val Arg 305 310 315 320 Gln Ser Phe Leu Lys Ser Pro Ser Ser Met Val Val Ser Phe Gly Pro 325 330 335 Glu Arg Cys Ile Pro Pro Ser Ile Leu Lys Gly Leu Ser Ser Ile Val 340 345 350 Thr Thr Ala Ala Glu Tyr Gln Pro Ser Tyr Leu His Arg Asp Pro Glu 355 360 365 Leu Cys Asn Pro Asn Glu Ile Ala Val Ile Gly Met Ser Cys Lys Val 370 375 380 Ala Gly Ala Asp Asp Val Asp Glu Phe Trp Asp Leu Leu Cys Lys Ala 385 390 395 400 Glu Ser Gln His Gln Glu Val Pro Lys Glu Arg Phe Gly Phe Glu Ser 405 410 415 Ala Phe Arg Glu Val Asp Pro Thr Arg Lys Trp Tyr Gly Asn Phe Ile 420 425 430 Asn Glu His Asp Cys Phe Asp His Lys Phe Phe Lys Lys Ser Ala Arg 435 440 445 Glu Ile Ala Ala Thr Asp Pro Gln Gln Arg Gln Met Leu Gln Val Ala 450 455 460 Tyr Gln Ala Val Glu Gln Ser Gly Tyr Phe Thr Thr Pro Lys Ser Asp 465 470 475 480 Lys Asp Arg Lys Ile Gly Cys Tyr Ile Gly Val Cys Ala Ala Asp Tyr 485 490 495 Glu Tyr Asn Val Ala Cys His Pro Pro Asn Ala Phe Met Ala Thr Gly 500 505 510 Asn Leu Lys Ser Phe Val Ala Gly Lys Ile Ser His Trp Phe Gly Trp 515 520 525 Thr Gly Pro Gly Leu Cys Ile Asp Thr Ala Cys Ser Ser Ser Leu Val 530 535 540 Ala Val His Gln Ala Cys Gln Ala Ile Leu Thr Gly Asp Cys Thr Ala 545 550 555 560 Ala Leu Ala Gly Gly Ala Asn Ile Ile Thr His Pro Leu Trp Tyr Gln 565 570 575 Asn Leu Ala Ala Ala Ser Phe Leu Ser Pro Thr Gly Gln Cys Lys Pro 580 585 590 Phe Asp Ala Ser Ala Asp Gly Tyr Cys Arg Gly Glu Gly Phe Ala Ala 595 600 605 Val Phe Leu Lys Lys Met Ser Ala Ala Ile Ala Asp Gly Asp Met Ile 610 615 620 Ile Gly Ser Ile Lys Ala Thr Ala Val Asn Gln Asn Gln Asn Cys Thr 625 630 635 640 Pro Val Phe Val Pro Asn Ala Pro Thr Leu Ser Asp Leu Phe Arg Asp 645 650 655 Val Leu Asp Arg Ser Gln Leu Thr Ala Asn Gln Ile Thr Val Val Glu 660 665 670 Ala His Gly Thr Gly Thr Gln Val Gly Asp Pro Ala Glu Tyr Glu Ser 675 680 685 Ile Arg Asn Val Leu Gly Gly Pro Ser Arg Ser Thr Pro Leu Leu Phe 690 695 700 Gly Ser Val Lys Gly Leu Val Gly His Thr Glu Cys Thr Ser Gly Ala 705 710 715 720 Val Ser Leu Val Lys Thr Leu Leu Met Gln Gln His Glu Ala Ile Pro 725 730 735 Pro Gln Pro Ser Phe Asp Arg Leu Asn Pro Glu Ile Pro Val Ser Glu 740 745 750 Ser Asp Asn Met Gln Ile Ala Thr Arg Phe Ser Pro Trp Thr Ala Glu 755 760 765 Tyr Arg Ala Ala Leu Ile Asn Asn Tyr Gly Ala Cys Gly Ser Asn Ala 770 775 780 Ser Met Val Val Ala Gln Ala Pro Arg Thr Glu Gln Lys Arg Ser Ala 785 790 795 800 Thr Arg Arg Thr Ser Val Val Leu Asp Tyr Pro Phe Arg Leu Cys Gly 805 810 815 Ser Asp Asp Arg Ala Leu Arg Ala Tyr Ser Glu Arg Leu Val Arg Phe 820 825 830 Ile Ala Ser Gly Ile Lys Asp Gly Ile Ser Val Ala Asp Leu Ala Phe 835 840 845 Asn Val Cys Arg Gln Ser Asn Pro Thr Leu Asp Arg Ser Leu Ala Phe 850 855 860 Ala Cys Arg Thr Thr Gln Glu Val Glu Glu Lys Leu Arg Ala Phe Val 865 870 875 880 Ala Gly Asn Gln Gly Leu Ile Ser Ala Ser Arg Ser Lys Thr Pro Arg 885 890 895 Glu Val Ile Leu Cys Phe Gly Gly Gln Ile Ser Asn Tyr Val Gly Leu 900 905 910 Asp Arg Glu Val Tyr Asp Asn Val Ala Leu Leu Arg Lys His Leu Ala 915 920 925 Ile Cys Asp Ala Ala Cys Arg His Leu Gly Val Asp Ser Ile Phe Pro 930 935 940 Gly Ile Phe Gln Lys Ser Pro Ile Ser Asp Pro Val Lys Leu Gln Thr 945 950 955 960 Ile Leu Phe Ser Thr Gln Tyr Ser Ser Ala Lys Ala Trp Met Asp Ser 965 970 975 Gly Val Arg Pro Val Ala Ala Val Gly His Ser Phe Gly Glu Leu Thr 980 985 990 Ala Leu Cys Val Thr Gly Ile Leu Ser Leu Ala Asp Ala Met Lys Met 995 1000 1005 Ile Val Gly Arg Ala Thr Val Ile Arg Asp Phe Trp Gly Glu Asp Lys 1010 1015 1020 Gly Ser Met Ile Ala Val Glu Ala Asp Glu Asn Arg Val Gln Arg Leu 1025 1030 1035 1040 Leu Ala Glu Ala Ala Lys Gln Cys Glu Leu Ile His Val Arg Ala Pro 1045 1050 1055 Thr Ile Ala Cys Val Asn Gly Pro Thr Ser Tyr Thr Leu Ala Gly Pro 1060 1065 1070 Val Lys Ser Ile Asp Ile Val Thr Glu Val Ile Ser Arg Leu Ser Asp 1075 1080 1085 Ser Gly Pro Ser Ile Arg Ser Lys Arg Leu Lys Val Thr Asn Ala Phe 1090 1095 1100 His Ser Thr Leu Val Glu Pro Leu Met Glu Glu Leu Glu Lys Val Gly 1105 1110 1115 1120 Gln Gln Leu Thr Phe Asn Ala Pro Thr Ile Gln Leu Glu Arg Ala Ile 1125 1130 1135 Glu His His Ser Asp Ala Thr Leu Thr Ser Asp Tyr Val Pro Asp His 1140 1145 1150 Met Arg Asn Pro Val Tyr Phe Asn Gln Ala Val Gln Arg Leu Ala Gln 1155 1160 1165 Gln Tyr Pro Asp Ser Val Trp Leu Glu Ala Gly Ser Asn Ser Thr Ile 1170 1175 1180 Thr Ser Met Ala Ser Arg Ala Leu Gly Ser Pro Lys Ser Leu His Phe 1185 1190 1195 1200 Gln Ala Val Asn Ile Thr Ser Asp Asp Ser Trp Ser Met Leu Ile Thr 1205 1210 1215 Ser Thr Leu Ser Leu Trp Lys Gln Gly Ile Ser Thr Asn Phe Trp Ala 1220 1225 1230 Tyr His Ala Lys Gln Thr Tyr Glu Tyr Asn Pro Val Leu Leu Pro Pro 1235 1240 1245 Tyr Gln Phe Glu Pro Ser Arg His Trp Met Glu Leu Lys Val Pro Ser 1250 1255 1260 Phe Met Ser Asn Gly Asn Val Gln Cys Gly Pro Arg Asp Glu Glu Glu 1265 1270 1275 1280 Pro Pro Lys Thr Leu Trp Ser Leu Ile Glu Ala Ser Asp Lys Val Ala 1285 1290 1295 Arg Phe Gln Ile Asn Thr Ala Ala Pro Lys Tyr Val Glu Leu Val Ser 1300 1305 1310 Gly His Val Ile Ala Asn Thr Ala Pro Ile Cys Pro Ala Thr Val Glu 1315 1320 1325 Val Asp Ile Val Val Glu Ala Leu Arg Ser Leu Arg Pro Asp Phe Met 1330 1335 1340 Asp Ser Asn Leu Gln Pro Gln Val Leu Ala Val Thr Asn Gln Ser Pro 1345 1350 1355 1360 Ile Cys Ile Asp Pro Asn Arg Ser Val Trp Leu Glu Cys Gln Ala Met 1365 1370 1375 Asp Ser Asn Leu Val Trp Glu Trp Arg Ile Val Ser Asp Ser Leu Gln 1380 1385 1390 Glu Pro Gly Thr Ser Ser Ser Ala His Val Leu Gly Lys Leu Ala Phe 1395 1400 1405 Leu Ser Gly Gln Asp Glu Val Lys Gln Gln Glu Ser Glu Phe Met Arg 1410 1415 1420 Leu Glu Arg Leu Ile Gly His Gln Arg Cys Val Asp Leu Leu Asn Thr 1425 1430 1435 1440 Thr Glu Ala Asp Asp Ile Ile Gln Gly Arg Asn Ile Tyr Thr Thr Phe 1445 1450 1455 Ala Gly Val Val Asp Tyr Gly Glu Gln Tyr Arg Gly Leu Lys Lys Ile 1460 1465 1470 Val Gly Lys Gly Leu Glu Ser Ala Gly Arg Val Gln Lys Lys Pro Ser 1475 1480 1485 Glu Glu Ser Trp Leu Asp Ala His Leu Gly Asp Cys Phe Ser Gln Val 1490 1495 1500 Gly Gly Ile Trp Val Asn Cys Met Thr Asp His Asn Pro Asp Asp Met 1505 1510 1515 1520 Phe Ile Ala Thr Gly Phe Glu Lys Trp Val Arg Ser Pro Ala Leu Arg 1525 1530 1535 His Gly Gln Pro Arg Pro Glu Ile Trp Asp Val Leu Ala Cys His His 1540 1545 1550 Arg Ser Ser Glu Gln Thr Tyr Leu Thr Asp Ile Phe Ile Phe Asp Ala 1555 1560 1565 Glu Gln Gly Ala Leu Thr Glu Val Ile Leu Gly Ile Asn Tyr His Lys 1570 1575 1580 Val Ala Lys Ala Ser Met Ser Lys Ile Leu Ser Arg Leu Ser Gly Thr 1585 1590 1595 1600 Glu Ala Ala Pro Ser Ser Ser Thr Arg Ala His Pro Thr Ser Ser Ser 1605 1610 1615 Ser Pro Arg Leu Pro Gly Pro Phe Val Pro Glu Asp Lys Ser Gln Asn 1620 1625 1630 Glu Thr Gln Thr Ala Gly Thr Asn Ala Val Ala Lys Lys Lys Ser Glu 1635 1640 1645 Lys Ser Ala Gln Gln Asn Val Leu Asp Lys Thr Arg Ala Leu Leu Ala 1650 1655 1660 Glu Ile Ser Gly Leu Glu Pro Ser Glu Ile Glu Ala Glu Thr Gly Leu 1665 1670 1675 1680 Ala Asp Ile Gly Ile Asp Ser Leu Met Gly Met Glu Leu Ala Arg Asp 1685 1690 1695 Leu Glu Ala Leu Phe Lys Cys Pro Leu Leu Gly Asp Glu Leu Ala Asn 1700 1705 1710 Val Thr Thr Phe Gln Gly Leu Val Glu Tyr Val Gln Ser Ala Val Gly 1715 1720 1725 Val Pro Ala Asn Gly Asp Glu Pro Asp Asn Thr Asn Ala Asp Glu Val 1730 1735 1740 Phe Glu Glu Asp Asn Leu Ala Ala Ser Pro Ser Ser Ser Ser Ser Ser 1745 1750 1755 1760 Thr Asn Leu Thr Glu Asp Ser Ser Leu Asp Pro Thr Glu Thr Thr Thr 1765 1770 1775 Asn Ile Ser Ser Tyr Pro Gly Gln Thr Lys Thr Glu Lys Pro Ala Met 1780 1785 1790 Pro Pro Ala Ser Ser Lys Thr Leu Glu Leu Ser Pro Ala Trp Val Leu 1795 1800 1805 Glu Ala Phe Glu Glu Ser Lys Gln Leu Thr Asp His Phe Ile Glu Gln 1810 1815 1820 Tyr Arg Cys Ala Asn Tyr Val Asp Thr Ile Leu Pro Lys Gln Thr Gln 1825 1830 1835 1840 Leu Cys Val Ala Leu Thr Val Glu Ala Phe Glu Lys Leu Gly Cys Pro 1845 1850 1855 Ile Arg Ser Ala Val Ala Gly Gln Lys Leu Glu Arg Ile Leu His Ile 1860 1865 1870 Pro Lys His Ala Gln Leu Ala Gln Tyr Leu Tyr Arg Leu Leu Ser Ala 1875 1880 1885 Asp Ala Arg Leu Ile Asn Leu Thr Glu Asp Gly Arg Ile Thr Arg Thr 1890 1895 1900 His Met Ala Leu Pro Lys Pro Ser Asp Gln Ile Leu Gln Asp Leu Leu 1905 1910 1915 1920 Arg Leu Tyr Pro Asp His Glu Trp Ala Asn Arg Leu Ala Ala Phe Thr 1925 1930 1935 Gly Ala Arg Leu Ala Glu Val Leu Lys Gly Glu Thr Asp Gly Leu Gly 1940 1945 1950 Leu Ile Phe Gly Thr Asp Glu Gly Arg Glu Leu Val Ala Gly Leu Tyr 1955 1960 1965 Gly Asp Ser Leu Leu Asn Lys Leu Ser Tyr Arg Gln Met Glu Asp Ile 1970 1975 1980 Ile Thr Arg Leu Ala Ser Arg Ile Pro Arg Asp Ser Gly Pro Leu Lys 1985 1990 1995 2000 Ile Leu Glu Met Gly Ala Gly Thr Gly Gly Thr Thr Arg Gly Met Ala 2005 2010 2015 Pro Leu Leu Ala Arg Leu Gly Ile Pro Val Glu Tyr Thr Phe Thr Asp 2020 2025 2030 Leu Ser Gly Ser Phe Val Ala Ala Ala Arg Lys Lys Tyr Gln Lys Glu 2035 2040 2045 Tyr Pro Phe Met Lys Phe Gln Val His Asp Ile Glu Lys Pro Pro Ser 2050 2055 2060 Asp Gln Leu Arg His Ser Gln His Ile Val Ile Ala Ser Asn Ala Ile 2065 2070 2075 2080 His Ala Thr His Ser Leu Thr Asp Ser Ser Arg His Val Arg Glu Phe 2085 2090 2095 Leu Lys Thr Asp Gly Phe Leu Met Ile Val Glu Met Thr Gln Pro Val 2100 2105 2110 His Trp Val Asp Ile Ile Phe Gly Leu Phe Asp Gly Trp Trp Leu Phe 2115 2120 2125 Ala Asp Gly Arg Asp His Ala Ile Ala Ser Ala Gly Trp Trp Glu Lys 2130 2135 2140 Val Phe Gln Ser Val Gly Tyr Gly Gln Val Asp Trp Thr Asp Gly His 2145 2150 2155 2160 Arg Pro Glu Val Gln Ile Gln Arg Val Ile Ile Ala Leu Ala Ser Gly 2165 2170 2175 Pro Arg Tyr Gly Arg Gln Pro Leu Pro Pro Ala Pro Pro Pro Asn Leu 2180 2185 2190 Val Pro Gly Ser His Ala Ser Arg Gln Ala Ala Val Asn Glu Tyr Leu 2195 2200 2205 Asp Lys Tyr Thr Lys Gly Phe Thr Leu Pro Ala Gln Thr Ser Asn Thr 2210 2215 2220 Asp Ile Ser Asn Ser Thr Ser Tyr Trp Glu Lys Gln Cys Val Leu Ile 2225 2230 2235 2240 Thr Gly Ala Thr Gly Ser Leu Gly Val His Leu Val Ala Ala Val Ala 2245 2250 2255 Ala Leu Asp Asp Val Gln Thr Val Ile Cys Leu Asn Arg Arg Ser Pro 2260 2265 2270 Met Asp Pro Asp Leu Arg Gln Gln Gln Ala Phe Glu Arg Arg Gly Ile 2275 2280 2285 Leu Leu Glu Ala Ala Ser Met Ser Lys Ile Arg Val Leu Gln Thr Asp 2290 2295 2300 Ser Ser Lys Pro Gln Leu Gly Leu Thr Asp Glu Val Tyr Ser Ser Leu 2305 2310 2315 2320 Val Thr Ser Thr Thr His Ile Ile His Asn Ala Trp Pro Met Thr Gly 2325 2330 2335 Lys Arg Pro Leu Ser Gly Leu Glu Gln Gln Phe Leu Val Met Arg Asn 2340 2345 2350 Leu Leu Asp Leu Ala Ala Gln Cys Ser Ser Thr Arg Pro Ala Asn Ala 2355 2360 2365 Pro Arg Ile Val Phe Gln Phe Ile Ser Ser Ile Ala Thr Val Gly Tyr 2370 2375 2380 Tyr Pro Leu Trp Ser Gly Gln Thr Leu Val Pro Glu Ala Arg Met Gly 2385 2390 2395 2400 Ile Glu Ser Val Leu Ala Asn Gly Tyr Gly Glu Ala Lys Tyr Val Cys 2405 2410 2415 Glu Gln Met Leu Asp Arg Thr Leu His Gln Tyr Pro Asp Arg Phe Arg 2420 2425 2430 Ala Met Ala Val Arg Leu Gly Gln Ile Ala Gly Ser Arg Thr Ser Gly 2435 2440 2445 Tyr Trp Asn Pro Met Glu His Leu Ser Phe Leu Phe Lys Ser Ala Gln 2450 2455 2460 Thr Leu Gln Val Phe Pro Asp Phe Thr Gly Asp Phe Asp Leu Leu Leu 2465 2470 2475 2480 Arg Ser Thr His Ser Asn Ser Thr Thr Asp Gln Pro Ile Tyr His Ile 2485 2490 2495 Asp Asn Pro Val Arg Gln Ser Trp Ser Glu Met Val Pro Val Leu Ile 2500 2505 2510 Asp Ala Leu Gly Ile Pro Ala Gln Asn Val Ile Pro Phe Ala Asp Trp 2515 2520 2525 Val Cys Arg Val Arg Ala Phe Pro Gly Gln Val Glu Trp Asp Asn Pro 2530 2535 2540 Ser Ala Leu Leu Ile Asp Phe Leu Asp Asp His Phe Leu Arg Met Ser 2545 2550 2555 2560 Cys Gly Gly Leu Leu Leu Asp Thr Lys Arg Ala Cys Glu His Ser Pro 2565 2570 2575 Thr Leu Ala Ala Val Gly Pro Val Thr Ala Glu Leu Ala Arg Lys Tyr 2580 2585 2590 Ile Gln Ser Trp Lys Glu Met Gly Phe Leu Asn Pro 2595 2600 <210> 4 <211> 2473 <212> PRT <213> Aspergillus niger <400> 4 Met Ala Ser Glu Thr Ala Ser Ala Met Asp Pro Gln Tyr Arg Met Glu 1 5 10 15 Leu Glu Val Val Tyr Glu Ala Leu Glu Ser Ala Gly Ile Pro Met Glu 20 25 30 Ser Ile Lys Gly Thr Asn Thr Ser Val Tyr Gly Gly Val Met Phe Arg 35 40 45 Asp Tyr His Asp Thr His Ser Arg Asp Leu Asp Thr Leu Pro Arg Tyr 50 55 60 Phe Met Thr Gly Asn Ala Ala Thr Met Ala Ser Asn Arg Ile Ser His 65 70 75 80 Phe Tyr Asp Leu Arg Gly Pro Ser Met Thr Val Asp Thr Gly Cys Ser 85 90 95 Thr Ser Leu Thr Ala Leu His Leu Ala Cys Gln Asn Leu Arg Ser Gly 100 105 110 Glu Ser Asn Met Ser Ile Val Thr Gly Ala Ser Leu Met Ile Asn Pro 115 120 125 Asp Val Phe Leu Ser Met Ser Asn Ile Gly Phe Leu Ser Pro Asp Gly 130 135 140 Ile Ser Tyr Ala Phe Asp Ser Arg Ala Asn Gly Tyr Gly Arg Gly Glu 145 150 155 160 Gly Val Gly Ala Leu Leu Val Lys Arg Leu Asp Asp Ala Leu Arg Asp 165 170 175 Gly Asp Ser Ile Arg Ala Ile Ile Arg Glu Thr Gly Val Asn Gln Asn 180 185 190 Gly Lys Thr Pro Ser Ile Thr Ala Pro Gln Gln Ala Ala Gln Glu Ala 195 200 205 Leu Ile Arg Gln Cys Tyr Glu Arg Val Asn Leu Asp Pro Ala Gln Thr 210 215 220 Thr Tyr Val Glu Ala His Gly Thr Gly Thr Pro Ala Gly Asp Pro Leu 225 230 235 240 Glu Val Gly Ala Leu Ala Ala Ala Leu Gly Gly Ser Arg Ser Ala Glu 245 250 255 His Pro Leu Tyr Leu Gly Ser Ile Lys Ala Asn Ile Gly His Thr Glu 260 265 270 Ala Ala Ser Gly Val Ala Ser Ile Ile Lys Val Ala Leu Ala Leu Glu 275 280 285 Lys Gly Gln Ile Pro Pro Asn Thr Gln Leu Asn Thr Pro Asn Ser Glu 290 295 300 Leu Arg Leu Asn Asp Arg Asn Met Glu Val Pro Val Ser Thr Gln Arg 305 310 315 320 Trp Pro Val Gly Lys Gly Pro Arg Arg Ala Ser Val Asn Asn Phe Gly 325 330 335 Phe Gly Gly Ser Asn Ala His Ala Ile Leu Glu Ser Pro Pro Val Gly 340 345 350 Ser Thr Asn Gly Thr His Val Asn Gly Thr His Thr Thr Asn Gly Thr 355 360 365 Ala Gly Met Asn Gly Thr Thr Gly Glu Asn Gly Ala Asn Arg Thr Asn 370 375 380 Gly Gln Leu Lys Ser Asn Gly Pro Val Val Asn Gly Asn Lys Thr Asn 385 390 395 400 Ile Thr Lys Arg Glu Thr Pro Trp Val Phe Arg Leu Ser Ala Lys Asp 405 410 415 Ala Gln Thr Cys Gln Gln Met Ala Ala Asp Leu Ser Thr Tyr Ile Glu 420 425 430 Ser His Pro Pro Val Asp Glu Glu Ala Phe Leu Gly Arg Leu Ala Tyr 435 440 445 Thr Leu Gly Ser Arg Arg Ser Val Phe Ser Trp Thr Ala Ala Val Ser 450 455 460 Ala Arg Ser Leu Ala Glu Leu Thr Arg Ala Leu Asp Asp Asp Glu Arg 465 470 475 480 Leu Val Pro Ser Arg Ala Ala Pro Ser Leu Arg Leu Gly Trp Val Phe 485 490 495 Thr Gly Gln Gly Ala Gln Trp Tyr Ala Met Gly Arg Glu Leu Ile Ala 500 505 510 Thr Tyr Pro Val Phe Arg Ser Thr Ile Leu Glu Cys Asp Arg Tyr Met 515 520 525 Thr Glu Met Gly Ser Thr Trp Thr Leu Met Glu Glu Leu His Arg Glu 530 535 540 Glu Ser Thr Ser Gln Val Asn Asn Ile Val Tyr Ser Leu Gly Leu Ala 545 550 555 560 Thr Ala Ile Gln Ile Ala Leu Val Glu Leu Leu Trp Ser Trp Gly Ile 565 570 575 His Pro Thr Ala Val Thr Gly His Ser Ser Gly Glu Ile Ala Ala Ala 580 585 590 Tyr Ala Ser Lys Ala Leu Asp Met Lys Ser Ala Ile Gly Ile Ala Tyr 595 600 605 Leu Arg Gly Val Leu Ala Glu Lys Phe Asp Asp Lys Ile Leu Gly Lys 610 615 620 Gly Gly Met Met Ala Val Gly Leu Gly Arg Lys Pro Val Glu His Tyr 625 630 635 640 Leu Ser Arg Val Thr Ala Gly Tyr Cys Val Val Ala Cys Val Asn Ser 645 650 655 Gln Tyr Ser Val Thr Ile Ser Gly Asp Ile Pro Ala Ile Asp Gln Leu 660 665 670 Glu Gln Leu Leu Gln Glu Asp Gln Val Phe Ala Arg Arg Leu Arg Val 675 680 685 Asn Gly Ala Phe His Cys Glu Gln Met Arg Pro Met Ala Asp Leu Phe 690 695 700 Asp Trp Ser Leu Arg Tyr Leu Leu Thr Pro His Pro Asp Phe Gly Ser 705 710 715 720 Val Leu Phe Ser Ser Pro Lys Thr Gly Ser Arg Ile Gln Asp Gly Thr 725 730 735 Ile Leu Ala Thr Ser Ser His Trp Val Gly Asn Met Leu Gln Ala Val 740 745 750 Glu Phe Glu Ser Ser Phe Arg His Met Cys Phe Gly Asp Pro Ser Pro 755 760 765 Lys Gly Ala Lys Gly Thr Gln Asp Val Asp Leu Val Leu Glu Ile Gly 770 775 780 Pro His Gly Ala Leu Gly Gly Pro Ile Gln Gln Leu Met Thr Leu Pro 785 790 795 800 Glu Phe Glu Gly Ser Gly Ile Ser Tyr Leu Pro Thr Leu Val Arg Lys 805 810 815 Gln Asp Ala Val Phe Ala Met Gln Arg Leu Ala Ile Asp Leu Thr His 820 825 830 Arg Gly Tyr Pro Val Asp Leu Asn Ala Val Asn Phe Pro His Gly Thr 835 840 845 Leu Ser Leu Ser Ile Leu His Asp Leu Pro Ser Tyr Pro Trp Asn His 850 855 860 Ser Thr Arg Tyr Trp Leu Glu Pro Arg Arg Asn Arg Ala Asp Arg Gln 865 870 875 880 Arg Gln Ala Pro Pro Ser Asp Leu Val Gly Tyr Ser Gln Pro Ser Ile 885 890 895 Thr Pro Leu Ala Arg Thr Trp Arg His Ile Ile Arg Leu Ser Asp Leu 900 905 910 Pro Trp Leu Gly Asp His Arg Val Gln Ser Ser Ile Val Phe Pro Gly 915 920 925 Ala Gly Leu Val Ser Met Ala Ile Glu Gly Met Arg Gln Val Ala Ala 930 935 940 Gly Arg Gln Gln Thr Val Ser Ala Tyr Glu Leu Arg Asp Val Asp Ile 945 950 955 960 Ala Lys Ala Leu Thr Val Pro Glu Ala Asp Glu Gly Val Glu Val Gln 965 970 975 Leu Asn Ile Arg Pro Cys Asp Glu Gln Met Leu Gly Thr Lys Asp Trp 980 985 990 Leu Ala Phe Gln Ile Phe Ser Val Ser Gly Asp Ser Arg Trp Thr Glu 995 1000 1005 His Cys Ser Gly Arg Ile Ser Val Ile Thr Thr Ser Asp Ser Thr Pro 1010 1015 1020 Leu Pro Ser Ala Ile Pro Ser Gln Ser Glu Asp Leu Tyr Asn Arg Arg 1025 1030 1035 1040 Ile Asp Pro Arg Tyr Met Trp Ala Ala Met Arg Ser Val Gly Ile Tyr 1045 1050 1055 His Gly Pro Leu Phe Gln Asn Ile His Gln Val Leu Ala Lys Pro Ser 1060 1065 1070 Ala Ser Arg Thr Ile Phe Ala Ile Ala Asp Thr Ala Ala Val Met Pro 1075 1080 1085 Lys Lys Tyr Gln Thr Pro His Val Leu His Pro Thr Thr Leu Asp Ser 1090 1095 1100 Val Phe Gln Ala Ala Tyr Thr Leu Leu Pro Glu Ser Gly Ala Arg Leu 1105 1110 1115 1120 Pro Ser Ala Met Val Pro Arg His Ile Arg Ser Val Arg Val Ser Ala 1125 1130 1135 Gln Ile Ser Asn Ser Pro Ala His Asp Leu Glu Ala Tyr Ala Thr Leu 1140 1145 1150 Asn Arg Asp Tyr Asp Ala Gln Ser Phe Glu Thr Ser Leu Thr Val Val 1155 1160 1165 Asp Ala Lys Asp Gly Asn Ser Pro Val Leu Glu Val Asp Gly Leu Thr 1170 1175 1180 Cys Gln Ser Leu Gly Arg Ala Leu Asp Arg Glu Ala Asp Pro His Glu 1185 1190 1195 1200 Asn Glu Ile Cys Ser Arg Trp Glu Trp Ala Pro Asp Ile Gly Thr Leu 1205 1210 1215 Asp Ala Ala Ala Cys Lys Asp Arg Ile Arg Cys Ala Pro Glu Ala Ala 1220 1225 1230 Glu Ile Glu Thr Met Arg Asp Leu Arg Arg Ala Thr Ile Leu Tyr Ile 1235 1240 1245 Leu Asp Ile Val Ser Ser Leu Thr Val Ala Asp Val Gln Gln Leu Arg 1250 1255 1260 Gly His Leu Lys Lys Phe Tyr Val Trp Met Val Glu Gln Leu Lys Lys 1265 1270 1275 1280 Ala Ser Arg Asn Gln Phe Ala Pro Asp Ser Ala Gln Trp Arg Asp Ile 1285 1290 1295 Ser Ala Ala Asp Lys Ala Ala Leu Tyr Glu Lys Val Gly Arg Thr Ser 1300 1305 1310 Val Asn Gly Glu Met Leu Cys Arg Leu Gly Pro Leu Gly Ala Ser Phe 1315 1320 1325 Leu Arg Gln Glu Met Ala Pro Leu Glu Val Met Leu Glu Asn Arg Leu 1330 1335 1340 Leu Phe Arg Tyr Tyr Leu Glu Ala Leu Lys Trp Asp Arg Ser Thr Arg 1345 1350 1355 1360 Gln Val Ser Glu Leu Val Arg Leu Cys Thr His Lys Asn Pro Arg Ala 1365 1370 1375 Lys Ile Leu Glu Ile Gly Ala Gly Thr Gly Gly Gly Thr Gln Val Ile 1380 1385 1390 Leu Glu Ala Leu Gly Lys Glu Asn Gly Ser Ser Thr Gly Ala Arg Phe 1395 1400 1405 Gly Arg Tyr Asp Phe Thr Asp Ile Ser Ala Gly Phe Phe Glu Ala Ala 1410 1415 1420 Lys Glu Arg Phe Gln Asp Trp Ala Asp Leu Met Asn Phe Gln Lys Leu 1425 1430 1435 1440 Asp Ile Glu His Asp Pro Val Ala Gln Gly Phe Glu Glu Gly Ser Tyr 1445 1450 1455 Asp Val Val Ile Ala Cys Gln Val Leu His Ala Thr Lys Ser Met Asp 1460 1465 1470 Arg Thr Leu Thr His Val Arg Lys Leu Leu Lys Pro Gly Gly Lys Leu 1475 1480 1485 Ile Leu Met Glu Thr Thr Arg Asp Glu Leu Asp Val Phe Phe Ala Phe 1490 1495 1500 Gly Leu Leu Pro Gly Trp Trp Leu Ser Glu Glu Glu Glu Arg Arg Thr 1505 1510 1515 1520 Thr Pro Ser Leu Thr Leu Pro Phe Trp Asn Gln Val Leu Ser Arg Asn 1525 1530 1535 Gly Phe Ala Gly Leu Asp Leu Glu Val His Asp Cys Asp Ser Glu Glu 1540 1545 1550 Phe Tyr Ala Phe Ser Thr Ile Leu Ser Thr Ala Gln Ala Pro Ala Leu 1555 1560 1565 Ser Ile Thr Ser Pro Val Thr Ile Val Thr Gly Thr Ser Pro Pro Pro 1570 1575 1580 Thr Ser Trp Met Ser Glu Leu Gln Thr Ala Val Ala Ala His Ile Gly 1585 1590 1595 1600 Cys Gln Pro Val Ile Ala Thr Leu Glu Thr Val Thr Pro Gln Gly Asn 1605 1610 1615 Ile Cys Ile Phe Leu Gly Glu Ala Asp Glu Pro Leu Leu Asp His Val 1620 1625 1630 Ser Asn Pro Val Glu Phe Asp Arg Ile Ile His Leu Ala Thr Arg Cys 1635 1640 1645 Lys Gly Leu Leu Trp Ile Thr Arg Gly Gly Ser Leu Asp Val Asp Lys 1650 1655 1660 Pro Ala Met Ser Leu Ser Gln Gly Leu Leu Arg Thr Leu Lys Ser Glu 1665 1670 1675 1680 Tyr Gln Gly Lys Ser Phe Val Ser Leu Asp Val Asp Pro Arg Arg Ser 1685 1690 1695 Pro Trp Thr Ala Glu Val Val Gln Ala Ile Ser Gln Ile Phe Pro Ala 1700 1705 1710 Ser Phe Ser Glu Thr Thr Asp Pro Ala Thr Cys Glu Phe Glu Tyr Ala 1715 1720 1725 Glu Arg Asp Gly Val Leu His Ile Pro Arg Thr Val Lys Asp Ile Pro 1730 1735 1740 Met Asn Arg Asn Ile Phe Pro Glu Ser Asp Thr Thr Glu Lys Thr Ile 1745 1750 1755 1760 His Cys Arg Phe Arg Asp Ala Ala Arg Pro Leu Arg Met Lys Ile Gly 1765 1770 1775 Thr Pro Gly Leu Ile Asp Thr Leu Val Phe His Asp Asp Leu Asp Ala 1780 1785 1790 Lys Ser Asp Pro Leu Pro Ala Asp Trp Ile Glu Phe Asp Pro Thr Ala 1795 1800 1805 Phe Gly Leu Asn Phe Arg Asp Val Met Val Ala Met Gly Gln Leu Glu 1810 1815 1820 Ala Asn Ala Ile Met Gly Phe Glu Cys Ala Gly Thr Ile Val Arg Leu 1825 1830 1835 1840 Gly Ala Thr Ala Ala Ala Lys Gly Phe Ala Val Gly Asp Arg Val Cys 1845 1850 1855 Thr Leu Leu Arg Gly His Trp Ala Thr Arg Pro Arg Ala Pro Trp Thr 1860 1865 1870 Ser Val Met Arg Ile Pro Gln His Leu Ser Asp Gln Glu Ala Ala Ser 1875 1880 1885 Phe Pro Thr Val Phe Ala Thr Ala Tyr Ile Ala Leu His Glu Thr Ala 1890 1895 1900 Arg Leu Gln Arg Gly Glu Ser Ile Leu Ile His Ala Ala Thr Gly Gly 1905 1910 1915 1920 Val Gly Gln Ala Ala Ile Gln Leu Ala Gln Leu Ile Gly Ala Glu Ile 1925 1930 1935 Tyr Ala Thr Ala Ser Thr Pro Ala Lys Arg Gln Leu Leu His Glu Thr 1940 1945 1950 Tyr Gly Ile Pro Glu Asn Asn Ile Phe Ser Ser Arg Asp Pro Ser Phe 1955 1960 1965 Ala Thr Asp Val His Leu Arg Thr Asp Gly Arg Gly Val Asp Val Val 1970 1975 1980 Leu Asn Ser Leu Ala Gly Arg Leu Leu Gln Glu Ser Phe Asn Cys Leu 1985 1990 1995 2000 Ala Glu Phe Gly Arg Met Val Glu Ile Gly Lys Arg Asp Leu Glu Gln 2005 2010 2015 His Ser Gly Leu Asp Met Tyr Pro Phe Thr Arg Asn Val Ser Phe Ser 2020 2025 2030 Ser Val Asp Leu Leu Thr Trp Gln Ser Arg Arg Gly Ala Asp Ile Ser 2035 2040 2045 Cys Val Leu Gln Ser Leu Ser Lys Leu Leu Gly Glu Lys Lys Ile Met 2050 2055 2060 Pro Val Tyr Pro Leu Thr Leu Tyr Pro Ile Thr Gln Ile Glu Lys Ala 2065 2070 2075 2080 Phe Arg Thr Met Gln Thr Gly Gln His Met Gly Lys Ile Ile Ile Ser 2085 2090 2095 Val Gly Glu Gln Asp Thr Val Pro Val Val Glu Arg Pro Pro Pro Phe 2100 2105 2110 Ser Leu Arg Ser Asp Ala Ser Tyr Val Ile Val Gly Gly Leu Gly Gly 2115 2120 2125 Ile Gly Arg Val Leu Cys Glu Trp Met Met Ala Arg Gly Ala Arg His 2130 2135 2140 Leu Ile Ile Ile Ser Arg Asn Ala Arg Pro Gly Pro Phe Val Thr Glu 2145 2150 2155 2160 Leu Glu Gln Gln Gly Cys Glu Val Arg Thr Leu Ala Cys Asp Ile Ala 2165 2170 2175 Ala Glu Asp Gln Leu Ala Ala Ala Leu Ala Gln Cys Ala Asp Met Pro 2180 2185 2190 Pro Ile Lys Gly Val Ile Gln Gly Ala Met Val Leu Lys Asp Thr Val 2195 2200 2205 Leu Glu Gln Met Thr Val Gly Asp Phe Glu Ala Ala Val Arg Pro Lys 2210 2215 2220 Ala Gln Gly Ser Trp Asn Leu His Gln Gln Leu Gly Asp Val Asp Phe 2225 2230 2235 2240 Phe Ile Met Leu Ser Ser Leu Met Gly Val Met Gly Ala Ala Ser Gln 2245 2250 2255 Ala Asn Tyr Ala Ala Gly Gly Ala Phe Gln Asp Ala Leu Ala Thr Tyr 2260 2265 2270 Arg Arg Asn Arg Gly Leu Pro Ala Val Ser Leu Asp Leu Gly Ile Val 2275 2280 2285 Arg Ser Val Gly Phe Val Ala Gln Thr Asp Gly Val Gln Glu Arg Leu 2290 2295 2300 Val Gln Met Gly Val Thr Ser Leu Ser Glu Glu Thr Val Leu Arg Ile 2305 2310 2315 2320 Leu Glu Gln Ala Ile Thr His Pro Thr Gly Pro Pro Gln Ile Ile Thr 2325 2330 2335 Gly Ile Asn Thr Ala Pro Gly Lys His Trp Asp Glu Ala Ser Trp Ile 2340 2345 2350 Gln Asp Pro Arg Phe Ala Ala Leu Arg Tyr Arg Asp Ser Thr Gln Ala 2355 2360 2365 Gly Ser Ser Arg Ala Thr Thr Gly Thr Ala Lys Gln Gly Lys Ile Arg 2370 2375 2380 Asp Gln Leu Ala Glu Ile Ala Ser Pro Val Asp Ala Ala Ala Leu Ile 2385 2390 2395 2400 Cys Gln Glu Leu Met Gln Lys Leu Ala Ser Met Phe Gly Leu Val Val 2405 2410 2415 Glu Glu Met Ser Ala Thr Gln Asp Leu Ser Ser Tyr Gly Val Asp Ser 2420 2425 2430 Leu Val Ala Val Glu Leu Arg Asn Trp Leu Val Ala Gln Val Gly Ala 2435 2440 2445 Glu Val Ser Ile Phe Asp Leu Met Gln Ser Pro Ser Leu Glu Asp Leu 2450 2455 2460 Ser Leu Arg Val Ala Thr Lys Arg Thr 2465 2470 <210> 5 <211> 67885 <212> DNA <213> Aspergillus niger <400> 5 tcactcagtc tcaacaccat ctccgaggcg attggcaatc tccagtagcg aatccttcac 60 cttggcctgc cagtcgagga gctgcttctg ccgggccatg cgcgtgttat aaacgaactt 120 gatctgcagc gagtcaccga taacaatggc ttgcgtctca accagcgcgg tgcggaggag 180 accggaccca gatccagttg ccttgggctt cttcacatcc acataacgaa gtaccgaatc 240 gtcgcgctcc agctgctgat actggccaag gtagttcata gacaactcga aggggctgtg 300 atgttcaaag gcggcatggc catcgtcggt cagccagcgg gttgcccagt agggaactcc 360 attgcctggc acactggcgc gcgcatctcg cacagcctgc aggtcgcga caatatctcc 420 agatgggttt atcactaccg gagtggagga ggtaaaccat ccgatagtgt tagatacatc 480 aagatctcca ccaaactcat cccgaccatg actgtagata aagaccgcag gtacgggacg 540 gtcagtgaac gtcgcgctga aggcatggaa gatcgccgac aagaagatgt caagtggctt 600 agtctggtac tgatggttgc aatttcccat caagcgggcg gttgtcttgg tgtccagaga 660 gaactcctgg atagtttcgg tcacgaaaag gttcatggat tcctccatct gccagtaatc 720 gtacagtgga gcaggaatcg agaaaggtag tacatattcg ggagtccatt tctcatcagc 780 gtgctccttg agtttacggc accacaaggc aaatgagtcg gaacggtggg aggtgattgt 840 cccggtggat aagccttgtt gcatatcttg gagaagaatg cgccatgaaa cggtgtcgat 900 aaccatgtgg tgtacagtaa gtccaacgta ccggtctccg ttcggaagct tgaaaataat 960 cgcacgcatc aatggaccac gctgcaaatc gaagccagac tggttgggcg cagtgtatgg 1020 ttcaatagct tccaagtcag gcacttccaa ctcctggtac agatgactat tggaccggtc 1080 aaggagata tactgcatcc agcggcccga ttcatcgcgc tggaaccgag cacggagcat 1140 gggatgacgc tctactgttg catccacgac cggttttagg tcggaatcag aatgaaccga 1200 tagggaac gtagtgttgt aacgattgag atcgcagtgc tgtttcaggt cgaactccca 1260 1320 ttcgtccaaa ggtcgcactc tcggtgccag ttcggcgata gtgggacact gaatgagatc 1380 cttaactgcg agcgagagct tggcatcacg gcactgggcg ataacctgca tagcggagat 1440 ggaatcgcca ccaaggcttt ggaaggaccg ctccatactg atttgctctt cgggtcgatt 1500 tagcacggag gcccaaatgc gtctcagctc aaattcggca ggtgtactcg ggccgcggcc 1560 gctagctttg gatcctacgt tggcaatctg ctcatatgtg gcatcgtcca gggtctcgac 1620 ccatcgccgg accttcttcc ggtccagctt tccggaaggc aacaagggga tcgcgttcac 1680 tggtacccac agggccggca ccatgtaaat cggaagggct gcagatatgg tgttagcgat 1740 tctggtcacg accgccataa tcttgggtct cctgtcctcc ggcacgagag ttaatcgtcc 1800 ctcctcaaca gggagatcga agcctttgaa agaaacaacg gccaatagct gcttcttgaa 1860 tgggccctga gatgggagca aagtgagtgc ctggtggata ttctcatcac gctcgatatt 1920 gtactcgatc tcgcccaatt caacccgctg gccgcgaatc ttcacctgcc catcagctct 1980 gccgatgaaa tgaatcgtgc catctgcatt gtatcgcaca agatcaccag tgcggtaata 2040 ccgctcacgc ttgtgagatc ctggaacaaa gctcttctgt ttgacaaagg cggctgctgt 2100 tttctcagga tcacccaggt agcctctcgc cagaattgga cctgaaatca agagctcacc 2160 aacagcacca atgggaagta gcttcgaagg ttgcttggca tcaacaatcc agagcacacc 2220 accaacagca tgtccaatgt tggctggatc ggtagtaggt tgcaatcctg ggttagctgc 2280 tgctgccaca gagcactcac taggtccgta tccgttgtac agctgtaccc tgtcagccca 2340 gttgagaatg tcaccctttg agagagcttc gccgccaaga accaaagtct tcaatgaggg 2400 cacatcggac ggttgtaaaa gcttaaggac agatggcgta aggaatgtcc agttgatatt 2460 gagcacgttg atgcagtgct tgatgtcctt gctcaagtct tcgtccggca cgcaaatgca 2520 tgcaccaaac atgagtgatg tcatgatttc cagcagacta acgtcgaagc tgtaggaagc 2580 aaactgcagt actcgtgatg agggacccag acaggcaagc tttccttgag ccagggctcc 2640 agagcagaat gcagaatgct gcacaacaca tccttttggc ttgcctgtgc tgcccgaagt 2700 gaagataacg taggcatcat gatgggggcg ggctctcacg acaccttggt cgagatcgtc 2760 ttgttcttcg ttcagcttgt cgaggagatc ctggtcaaca ctgacaatgg ttggaataag 2820 gtcgttccag tttgaagcat atttcggaga tgcaagaaca acgttagcag cagtttgaat 2880 agaggcctcg cgtaggcgct ccaccggggt tttaggatcc agagggacga acgcagctcc 2940 agtacccaga acagcaaaca tggcaacgat agcccatgct gacttttcga acgtcaaagg 3000 cacgtaatcc ccggcgccaa taccaagcct ggtcaagtat cgagacaacg acagccacat 3060 ccgtccaaaa tcctgatagg ttatcttttc ggccatggtg tcaatcgcaa tagcatgagg 3120 ggtacgcgta atctgttgga gcacgaggtc cggtactgtg tacgatgaaa tctccggcac 3180 atcctgattc caagatacaa tctgcgacaa atcataccgg ccgataacat tgagctgatc 3240 aactcgggcc tctggatgtt cgaccacgga tttgaggatg gcagagaatg ttcccgatac 3300 ggatccagct cgggattctg agatacaatc agtccaatat ccgagtttga tgtccacgtt 3360 gtttggcgaa tcgacaacgt gaaccgtgat attgtactac ttcaggtcag ttattgcaaa 3420 gtagttcaga gtaaagtcat acctctgtcg gctcccgtac aatgagaccc tcgaagtcaa 3480 tattggtgat aacgtcgtca ctatcaacgt ctgtcattgt ctggaaggaa atgcctgtgt 3540 taaacagagg cattcctgcg agtccaagtg aatgttgaat ctcggcgagc gagcagtgct 3600 ggtgtgcgag agaatcaagg aactcttcct ggcaagcctt caaggcgtcc ttgataatag 3660 tggagtcctc cagcaatgcc ttgcaaatga ggacactgat agttggcccg accatgttga 3720 ctaggtctt gacaggtgca tctcttccgg atgccaagta accgaaacaa atatcatctc 3780 ggccgacata cacccgacag acgagtgccc agactgtctg tagaaggtc gagattgtga 3840 tactctggga gacacagaag tcacgcagaa cactggtagg tattcctagc tctacctgga 3900 cgtacttcag ctcaggtgta ccttcagcct cagctaattt gggaaagtag cagggctcga 3960 cttcggcaag atggttctgc cagaaagaaa tgctagcgga catgtcgttt gcagcaacat 4020 attggacgaa gtccctgtac ttagcgcctc cagtgggaga aagaagagtc cccgcataag 4080 ctcgcttcag gtcccttaga atcaatgact tggatacacc atcgacaatg gtatgactag 4140 catcgactcg gcagaacact ctatcatccg ctgtgacatt gattgagaac cgatgtggag 4200 ggacaacagg gttagtgcct agttcaatcg tctcctccag cacagatgca acatcgacat 4260 ctgagcaagg tttcaccgaa acctccgcccc tgggacgtct cagaacaatt tgagcgtatg 4320 gctccttgcc gccaaattcg tcggtgaata tagtcctcag ggcagggtga tggtcgacta 4380 ccttttgcca tgaggagagc aatctgccaa tatcaactgt gtcgtttgcg ccgcaagaga 4440 ctacctccca tccaatgaat acattgtaac tgccttgctt cctagcctgg ctcataagga 4500 gaccttgctg caacggagag cagggataca tatcttctat ctgatttggt ctaaggccaa 4560 ctcgtggaag gtgctgacgg ttgactctct caagggatgg atgatcgagt ttgagcaatg 4620 gaaagtccgc caaagtgaat tccttagctg tattggacgc gaggatctcg actgcttgcg 4680 tgatgttctg tctgtactgg tcggtaaata gtaatagctt ctgagcgctg acctgatcgg 4740 atccgactgt gattctgaac tgtccatctt cgagcagaca tgaaacctcg aaagttgtgg 4800 gaacgatggc gcccaagccg tgtcgccgag tgagggtggc ttgcttcaac acgatacctt 4860 ctgtaagtcg gctagacaag ccgtagttga agatgacatt ggaagagcca agcttgctac 4920 ctatccccct ctggcgaact ttgtccttag tatcccggat tagctgaaga gtgacttggt 4980 ccgagtgttc cacaggcaca tgcacgggtt gcagcgaaga gaattgaccg acatgacctc 5040 tcaagtgttt gctaagagtc gactcagtag cggttgtttc gacaatagct aaggtagagt 5100 ggtcattttc ttctagaatt tggtggtatg acaggatcaa ggtcgccaga aatatatccc 5160 ccgcgttggt gccaaatttt aggttgcacg gtccagtgag aagcccagta atctcggagc 5220 cgagttctag cgtctctgtg acatagtctc gatacccact aagagattca agggcgacag 5280 attgcaatgg accattcata ggctccatgc cagtatcgct cctgaggtga tcaagccagg 5340 catgagagaa ctcatctcgg gacaaaagct tactttccca gacgttttcc tctttttcat 5400 catttgactg aagcaagtaa tcgagatggc cgatgacagt acttagagac tgtgaattaa 5460 ggatagctgc atgcacgacg agagaaatgt gctgtgtgcc gcgaagagag aacagatcga 5520 tcatgagatt tgggttacca ctcacaccaa ctgacgcctg acgatgctgc ataatttctt 5580 gtgcttcttc caagctagag acatggttca gattcaagtg gtaaacgggg ttttcaagct 5640 cgtcgtggtc tctgaatgtt tgcaaccatg tcccgttctg ctcattgaag ccgagggtga 5700 gtgctgggta ggaagaaacg atactatcca aggcatcatt cagctgctgg atactaatct 5760 tgtgattgag ttccagtaac atggcgtggt gtatggcgct gttgcccgaa cgggacgcct 5820 cgaggaaggt gcgctgtagg tgtgatatgg gaaggtgatc tgtctcgctt ctccggagca 5880 catccgcaga gccagactcc tccagcttag aggcgatctc agtaacattg taccgcaaca 5940 gatcagctac taggacggtg actccgagtt tgcggaactc tgacatgacg gccatcgctt 6000 tgatagagtc accgcccaca gacaggaagc tgctgtgttt gccaatttgc tcctctggta 6060 cgttcagagc tttcgcccaa accttctgca cgagaatctc tgtcttggtc caagacgcct 6120 tgtcgccagg ttttgttgag ttacctatga tgtgttggaa ggcctcggtg ctcatatcag 6180 caacaaaatc cccaaccttc ttccggtcca tcttgcccga cgacatgagt ggaagactct 6240 ccaggacaat ccagatggtt gggaccatat acgtggggag caggtgtgac agcctttctg 6300 caatcgcttg ggtgtctgct gcggcctcgg catgatttac caggcgaata ctcgtgtgtg 6360 tctgtattga agcggttgta aaatgcttgt ggagagacac gataccaacc aagcgggatt 6420 tgcagtgacc agtgtaggga atgagagcca ttccgttacg gatgactgga tctgtccaga 6480 ggtggtgctc tacctcccct aattccatac gctgtccatt gagcttgact tggtggtcct 6540 tacgcccaat gaagtgaatc atgccgtccg catcatatcg taccagatca ccagtcttgt 6600 aaaagcgatt ccagattgag gatgtacccg gcaccatcca agccgggttt ctaaagaagg 6660 ctgatgaagt cttcacagga tccttgatat atccacgcga gacaatagga ccttggatga 6720 tcagctcacc tgtggcgcca acagggagaa gcacgttgtg gttttgcgga tcaacaatcc 6780 agcatacgcc accaattgct ctaccaatat tccgtgggct ggactggggg ttgagctgag 6840 tattggcagt ggctgcgacg gagcattcac ttggcccata gccattcatc aactggagcc 6900 ttcccgccca tgtcatcata tccgactctg acatagcttc gcctcccagt accagagtct 6960 tcaaaagcgg cagttgatca ggagtcagca ccttcaacac cgacggtgtc aagaaggccc 7020 agttgacacg aaagtcattc atggactgga caaagtctgc acgaccaagc tgctcgttcg 7080 ggatacagat gcaggcgccg aatataaggc ccgtgaggat ttccaaaata ctcacgtcaa 7140 agctgtatga agcgaactgg aatatccggg aatgaggtcc catgttggcc gcttcaccct 7200 gtcgcagaga cccactgcag aaggactggt gctggatcat cgtacctttg ggctttccag 7260 tgctcccaga ggtaaagatc acatatgcga gattctcagg cgtgacggtg actggagtaa 7320 gacacccgtc accactagtt attgcccga ctgtgtctttc attgactatc attgctttga 7380 cagaaaagtc atcgaagaga tcgtgatgtt tggctgtcgt caaaataatc ggtgcatcaa 7440 cttggctgac gatctctcgt agacgtgatg agggatgcgt cgggtcgagg ggaacaaagg 7500 caccaccggc tttgagtact gcaagcaacg ccacgacata ccatgcagac ttttcaaaac 7560 acagaggcac aatcacctcg gccttgattc ccagttccac caacaccgta gccagatggt 7620 ttgacagaga atccagctct gcatacgtca tctgtgcatc ccatgaatca atagcaggag 7680 cctgtggttg caaggacgct gtctttctaa ctagctcatg agcgcactca atcaccgcat 7740 taggcgcgtt ggaattccag ttgaaaatcc tcttatggct gagtgggccc aagtagcttg 7800 cactcgaaag ctcagtctct gagtggttgt tgaccaattc gaatgtccga tggagagtat 7860 cggccaaatg gactgcaagg tcctcagata ctgtagacgg agtatagagc atgtggatct 7920 caatgccatt tctactattt gtggcatgaa gttcgaggtc aaactatatg cggtaagtct 7980 ctatctttcg gttgctgatg aacaggtggg ttttgggtct tacatttggt gatggcgaga 8040 tgtgtttact atttaaactc tcttcacgaa agattacggc gctgcggaag gactggtagg 8100 attgagtttg ctgtttctcg tctggtgtct ccaaggctga tcgagcacta tgcaatgttg 8160 ccttcactgc ttggagaagg tcctcaactt tgcggaactc gtccaactcg aacaatagat 8220 ctgtggttat cgttagcttg tgctgatttt ggggctctgc atacaatact cactcgtgga 8280 gctggaacca gttgctgtct cgtgatggta ttcaaaatgc acgccttcgg tattatagca 8340 acgaagggca aggccaaagg ctgtgcagag tactccagat acctctgctg tgtctcctgt 8400 cttcacttca acaggcaaaa agccgtctat cgaagcaggt tctcttgtga ctatggtggt 8460 gtcatctttg atgctcttga aagacttgga agggatgcta agatgagatg gttgaggagt 8520 ggataaagcg atcggagcca ttttgcagct gattgatgag tgaaatgtcg gcttgcaaac 8580 tgatagatgc agactgcagg ccaccttaat taaggaaagt tatggaatga aatgaaagct 8640 tctcttccgg gtgatatgtc catgtcgaga aagccttgaa caaagatgtt cggccttcga 8700 aggcttagat cgaacccggg agccaatact ttgctcccct gtcagcttcg tgttgtaggt 8760 ctcaagcaac aggggctgaa ttgatctatc gtgcgactga agttgatacg atccttctgc 8820 atgccctcat cgcgtaattg tcgagacgtc tgtatagacc tgtctcga taccacgtct 8880 gataccaaac cgtcatgtaa taccatgtaa cgctggtacg gtaatggtga cggtaatgat 8940 actgcaatcg gggggagatg tcagacttga ccggaacggg aggacccacc ggccacatgg 9000 atagcaaaga tccttcctgc attgttcaag ccttcgcacg tcgcgccgca acgccgatcg 9060 tgtggggtcc aaaacatggg agtgcaagag ccgctgcagg atccgacctg caccaacggc 9120 gtggttagat ttacgcaccc gccaatgagc agcacccacg cttaaacagc gtctccacgg 9180 tcgtctgtgc ctatgattat gcccctcctg gacatacacg cgctcatgtt ggcgcattgc 9240 cccgactatg tcgggcacag acaggcagca gctctctgct gggattccca cctcaggatt 9300 tcggatagca ttcgagcacc ccaagcagtg ctatcgactt aggcagtgct agtctacgat 9360 tgccagactg aatccgagca gtgtagggtc taaagcacaa actgtatcgc tttgctccag 9420 atggcacaca tccgcgcttg ctctcagtac tccgcagaac aggcgaagtc gcaatgaact 9480 gcaccatcgc ctctgaagat gctttcgggc ctcaagctcg ggcatgttat ggcggatacg 9540 actttacttt gttgttcgag gagtctgtga tgacgatcgg cccgatcgga cttttactcc 9600 tatgtctttt ccctagagtg gtctacctat acagaaagtc actgagagat ggaagtaaag 9660 atggcgctta tacagccaaa ctcgtaggtc acaattttta tcccactgta cccgagtcgt 9720 cgtcgcatcg ttgacttgtc gcagatcagc tacggatggt tcgctgccat ccaaacggct 9780 attctagtat cctggacatc gggcgatgaa cccacaacac gcacgactgt cccagctgcg 9840 gcgctcagtc tggtcgccgc gttgcgctc ggtcgcag cgcacttcga acacacgcgc 9900 tccgtgaaac cgccccttgt tatcgaaatt taccttctat tcactatcgc cttcgatgcg 9960 gttcgactac gcacgctatg gggtatggga tatcatggtc acatattcaa gttatcgact 10020 atcttcgtca tatggaaagt ggtgttgctc gtgcaggagg cctggcctcg ggagcttggt 10080 ttgttggaac agtcgtacag tcctgaagag aaggtcggct ggatcaatcg gcgcatgttt 10140 tggtgggtga acccgttgct gtttctaggt tccaagaaag atttgcaagc ggatgatctc 10200 ttcaccctta accgctccct gcaatctgaa gtatgttcga agagcttctc ggaggtttgg 10260 caaaatggta agcgtgtttc ccctttcctg ttctttgaaa ccattgccaa ctatgaacag 10320 caaagccggc aagaaagggc aaaaaaaatg gacttctttg gattatggct tgggagcaca 10380 aattggtcct tctgactgcg gttcttccgc ggttgtgctt cacagccttc accttcaccc 10440 agccgttcct tatcgacgct gtcgttaaat acctgcagac tgtgtcggag aatcacaacc 10500 gtaaagatgg atacggatta cttggtgcat atataattgt atacgtcggc ctgggagtga 10560 gtttgagtag atctggacta tcttgcagag tgctgatact ggctcagatt tccgaagcat 10620 gctatcagca tatcacgtac cgcactattg tcctcatgag gggttgcctt gtgccgctaa 10680 tctatgaaaa gacgctctct atggacccca aaaaggctga agaatatgcg ccagtaacgt 10740 tgatgagcgc tgacattgag aagatagctt tcggaatgcg ctacatgcat gaggcttggg 10800 gaaatattat tgagatagct ttggctctgt ggctgctata tcgggagctg aactatggag 10860 ggctgagtcc cattcttatt gccgtcggta agactcttca atggagcaga caagatgaag 10920 cttggtactg acattcaact agtttgcggg gctaccgctg ctacaatggc accagctgtc 10980 gggaggcggc aggcgacttg ggttgagggc atacagaaga gaatcgatgt cacaacatac 11040 atgatcaact caatgaagag cttgaagttg gaaggactga ccccttggtt catggacttt 11100 gtacagaatt tgagggtcaa ggaaatagaa tacgctaata gattcagatc ttttctcata 11160 tacgctgtca gcttgtgtta gtacattgtt cccattgttc atgattagct gctcactaac 11220 ctgcgctagc attcggaact tcagtcatat cgcccgtcgt cggcttcggc gtattcatcg 11280 ccttatcgcg atctgacaat ggcccggagc tgacgacctc aagggcattc accacattgt 11340 ctctgttttc agtccttcag aacccaatgt caatgctact gcaatctgtt cccaacctca 11400 tctcagccgt cggatctatc gagcgcgtgc gcgtgttctt gatgaccgag aacaccagag 11460 aggtgggctt ccttagctcc tttacctctg agcaaaccct gttcccagaa atggacaacc 11520 tccgggacat ggagatgaag gataccgaaa tggagaatgc agtcgaggca ttgaactgga 11580 gcgtgggatg ggtatctact cgcgaaccgg tcgtgaaacg aatgacattt aagatccggc 11640 cgtctacact tacgatcatc acgggcccaa ctggttgtgg aaaatcgaca ttacttgctg 11700 gtttgagggg agagacagcc gtggcaaagg ggtacatgag atgccgcttt ccctctgctg 11760 cattttgcag tcaggatcca tggctgcaga atggaaccat actgagtaat attctcggcc 11820 cggctacgta cgaaccgcga tggttccggg aggtcacgca ggcttctgga ttgaggcagg 11880 acttgaaatc gatgccgttg ggtgttcaga cggttgtggg aagtaaggga ttgtcattga 11940 gtgggggtca aaaacatcgg gtggtgggtt attcgtttcg acatagagct catggttgtc 12000 gaatactaac aacaacaatt acaggcactt gcaagggctc tttattcgcg acagcccttg 12060 ttgcttcttg atgacatatt cagtggattc gatgcagaga cagagaagct ggtcatcgca 12120 cggctatttg gccagagtgg cttttgtagg aaacacaacc tgaccactat ccttgcaacc 12180 cattccagta agtggaaaac ctgcgcactc aagaaggata gctactaatt tggctgttga 12240 agctcgcctt tcctctctctg cagactacac catcacactt gatagcaaag tccatttttc 12300 ggaaaggag ggatcgattg tttctagtct tcctgggcct agcggtggtg actttctctc 12360 agagcccata ttgagcgatg caagattgag ttggattgtg aggcagtcca accccgatgt 12420 accgcacgag cagcttgccg cacagcttga aacatctgat gctagccgcc ggacaggaga 12480 tgtgacgata tacaagtact acctggaaat ggtcggaata ctcaattctg tggttttctt 12540 tatcgcggtt gcgattttca cattcagtct tgcatttccg agtgagtgcc gcaatccgct 12600 atatctgatc aaagctcaca gatgcaggtg tatgggtcca atggtgggct gccgcaaacg 12660 aaaagcaccc atacaaggac ctcggcatgt atttgggtgt atatgctttc ctggcagtta 12720 tggcggagct gtcgttattc ttaggatgct gataagtcta tacgctgtct ctacaaaagg 12780 cgatggctaa cctaaagcac gcatatgatg agcaatatga ttccccgagc gtcaaggaag 12840 ttgcataaga tcctccttcg cacggtgcta aagtaaggtt ttttgaccat accgaagact 12900 tctgctgact cctgcagtgc tcccatatcc ttctttcaca caaccgattc tggtgtgacg 12960 accaacaagt aagatcctat cgctgtatga tgatatgaag ctcaagaggc attaggttca 13020 gtcaagatat gcaactggtg gacatggagc taccccttgc tcttgttgaa acaagcgtcg 13080 gtatgtccaa tatatcgttc tctggtgttg aatagatgac ctttgctaac ctaactaagc 13140 tctcatgtct gcaatagcac agctcctaat agtattcatc actggaaagt accttgcagc 13200 tatcattccg ctatgcctgg footattcta cttcctgcaa aagttctacc ttcggacatc 13260 acgacagcta cggttcatgg agcttgaagc caaatctccc ctctattcca acttcatgga 13320 gaccgtgaac ggactggtaa ccattcgtgc tttcggatgg cagtctaact ttcttacgaa 13380 tatatgttgc ctaacagatg catcgcaacg gccttattac ctgctcttcg ttatccagcg 13440 atggctcact ctcgtccttg acatggttgt ggcaggaatg gcgacgctga ttgttggcat 13500 tgctgtcggc gctcccgggt ccattggagc cggttctgca ggactggggt tgctgaatat 13560 catcaacgtg agcgaaagtt tgaagcagct aatttcgaat tggactgtgt tagaaacttc 13620 aattggagca gtgtcacgtg tcaggcagtt tcaggatcag gttcagcctg aagatagcc 13680 ggatgggcac attgaagttc cagcgcgctg gccggagaga ggcgacgtta agttcttcaa 13740 tgtttttgct tcttacaggt aatgcctccg cacggaattc agaagccata ctgactggta 13800 atttgataga cctgatgacc aacccgttct ccatgacgtg acattatcaa tagcagcggg 13860 tgagatggtc gctatttgcg gtcctagtgg caggtacgtt gccttggtct ctcctacaga 13920 tcgctacata atactaaggt cgtgaagtgg caagagctca ctagttcaac tcctcttccg 13980 ccttctcgaa cccgacaagg gtcagattac catcgacgga ctggacatta gctctatctc 14040 ttgccaggac atccggtcat ctctctcgtg catatcccaa gcagtcacca tccttcccgg 14100 tactgtacga caaaacatag accctcttgg gaaggaacct gacgagagta ttatcaatgt 14160 cttaaggaag gtcaagctat gggacattgt atcaacgcaa cttggtggct tggacggcct 14220 ggttcaggaa gaaagcttct cgcagggaca gaagcagctt cttcgcttag ctgctgctat 14280 gttgaggaag agcaaagttg ttgttctaga tgaagcgacg agtaggtatg tttatcatcg 14340 catctcaaat ctagtgatcg ggtatgcctt aaggtcgaat atagctgata ttgtgccgca 14400 cagtgtcgac cccgaaactg acgatctcat gcagcggctg attcgcacag cattcgcaga 14460 ttgcacagtc attgcagttg tgcatcgaat ccatacaatc cttgattttc acaaggtggt 14520 tgtcattgaa agtggacgca ttatcgagtg tggccctccg aaagagttgc tggccgaaaa 14580 tggcctgttc agccaacttt acggaaaagg aaccaccact accaccatag acatggaccc 14640 cgagaagtcc tggctcagat tttgagcgat tagaagctca cgagatttc cctttcgatg 14700 tggccttctc aaggtgctat tatcaggcgt tgtgtttata gacggtgctt ggatgtgatc 14760 ggtgatgttt gacgtcggtg catctactgc cgctcagtgg catacagcac cggcgatata 14820 ctgataatca catttctcat ttgttctgct gcattccata acatccccaa tgcgttttgc 14880 cgccttattc tgcttatgca gcagtgactc attaaacgga attaattcct tcataattc 14940 aatacttcc ggatcaacca ttccggctta tcaggccacg tctcccaacg gttgagcata 15000 aatataaga tttatgtttt ttctatacaa ttctctacat tagatcagca taacacttca 15060 ctttgggtgt atcaagacca tactcagcat ggcgtcccac tatctcagca cgtcgccgga 15120 taccaacatc catttcacca ttactaaagc acagtctcca tcttcaccta cgccccttct 15180 tctctttctc cactactggg gtggttcgag cgcaacatgg cacaagcaaa cctcacctac 15240 ttccccacac accctgagca acatatacaa cactgttaca gttgatcttc gcggctgggg 15300 tcagtccaca ggacctgcag actccggcgc cagctcgaaa gactactcca ttacgcccat 15360 ggcatctgat atcgtctcca tgctatcgca cctccaaagc accacctccc tcctcgacaa 15420 cggcgtgatc ctagtcggtc actccatggg agcaaaggtt acactggcta ctcttgccaa 15480 gctttccgac aatcaattat ccctcgtgaa gggcctcgtg cttgtcgctc cggcgccccc 15540 aactccattg gtactccctg ctgagatgag cgaacagcaa cgcaaagcat atgataacga 15600 aggctccgtt cggtggacgg tcgagaacgt tctatccagc gtacagaata ttcgggtta 15660 cgatatggag ttgatcgtca aggatagtct ggcaggaagt actcttgctc gggacggctg 15720 gattctacat ggtatgcaag aagatattac atctgcttta gatgaggtgt ctagtcggct 15780 tgagggccgg aaggttaagg tgggtgtgct tgctggggcg gatgacattg ttgagaacaa 15840 ggatcgggtc gagaaggagg ttgctggggc gcttaccaaa agaggctttg aggtgcagtt 15900 tgaggtacta gggggcgtaa agcatttgat tccattggag agcccggaga gcgtggctag 15960 ggcgattcgt tcgactgttg aatagtgttt atatttagat gtcttgactg aagaagacga 16020 gatccagtca gtattcttgt aatattcatt ctacccatgt atgtaaagcg aggtggcaat 16080 gcaaatctca ttgactttca cgtctttgag acttgaagct ctttcactaa tttgaaattt 16140 atatgtacga aagccacgtc tgggtgcata taagatgggg ctgtggttta gtggtataat 16200 attcccttag catgggagag gtccggggtt cgattccccg cagctccatt ttttgctcat 16260 attttttagt atcactaaga ggtacatgaa catgatttac acacatgcct taaacccata 16320 acactaaatg atgttttgg cgaagacact cgagactcat actcatgtgt ttctacaccc 16380 tacagtgtca tacaaggtat gttaacattt gtagccatca tcgatcctac tctcatgtta 16440 gggtgtgcca aagtcacccg acaatgaaat gttcccgaat cccagttctc cgcatcattg 16500 acgatactag gtttccgctt gtgatcagct ggtgtctgga tggaagattt tagtcaaggg 16560 actggctagc tcacaagctc ccgccaagaa actcagctgc ggagttgaaa agactgcccc 16620 tggatataca ataatacaat attcgcatta accaataagc tccttcacct tcttatcagc 16680 gactgtaaat gagtagccgc tgctgcacag gtggcaccat aagctcggaa atagagtttc 16740 gcaaagagca gcataagaca cgtgcatgcc acgtttcgaa gccgattagc gttggggctg 16800 gaatgttcca ggataattgc attaatcaat gtactaggta tctgtattaa tctcagtctc 16860 gaaattccac tcacatatga catcatgggc tgtcgccatc ggtcaggggc tgaataatc 16920 atccaaaagg atccgaggag agccggctgc agccgcaatt ggaggatctc ccgcaaggga 16980 ttcagttcct gactcaagag cagcgatcct cccctgatga taggctagga ttaccccgga 17040 gggatgtgtc cattctttgt ctggctccac tgaagcttcg cgttccaaca cttgaggcac 17100 tacatttgat acaccaattt gggacattta gcgccgttgt gtttggtatc catatgttat 17160 acctgtaaca attggtaata ctgttttacc aacaatacgt ttgtgtatgg aacgtcaaga 17220 acacccatca ccgtcacgta tgcagcctcc cattcagctc cgcctcgtcg gatcctgcaa 17280 catgcatttt tacgctgcat attagattat ccttctgttg tacactcggt gccaaatcgc 17340 tcggtcttgc cactgctgac cgagatctcc gatctggccc agtccgttat cgtgagatga 17400 tcgcattgcc aggcttatct ccccgtgcca ccgtgtgtgg tatgatatat cacttcacag 17460 ggcaactgtt ctgagtccag attctggagt gcagttgagt ctcctttcac tttattccct 17520 catgatgacc gtcaagacgc cgttgtcgag tttgagctcg gaatttgcgg gcctcactcc 17580 cgaaatctcc cggacgtctt ctgtcactgg cgacaaacgg tcattgatcg ctgcatatgg 17640 atctgttaat acgacgatct ccctacctac attacagctg ccgttggcaa accaattatg 17700 gaaggacttc agatcgaact tgccttcgtc ctccccagag aacgacttaa ctattaaaga 17760 aggactcgtc gccgagttca tcaacttctg catcacccgg tctagcgacg acactcattg 17820 tgacgacccc gcggcagtac tccagttcct cgaatctctg ttagaagctt ttgagcaaga 17880 ctttgttggt ggaaatgatg ttcactccgc cgtatcacac cttgacactc aacataaaac 17940 ccccataatc aaagcatact tcgcagcata cggaaccctc agtcgcaagc ctaggcgtca 18000 accttctatt ctcctcgagt ccgttacaga tggaccgtca cgtatatacg caatatttgg 18060 tggtcaagga aatgacgaga aatatttcgc ggacctaaaa gacacctacg cgacttacac 18120 cgacatcctc cgtcggtgtg tcttctccgg agccaggttg ttcgaggacc tcgctggggc 18180 atctgtattc aaaacccact ttcaacatgg cttcgcgctc atggactggc tggaaaagcc 18240 cgaatctgag ccagagacct cgtatcttct gtctgcaccc gttagcatgc ccctgatcgg 18300 tctcctccaa atgtgccact tcatcgtggc ctgtcaaaca atgggactct caccgagaga 18360 gatgcatggc tatttgtccg gtgtaacggg tcattcacag ggaattgtcg tagccctcgt 18420 ggtgtctgtt tccgatactt gggaagactt ctaccgacac tctctcgaag ccctccgtat 18480 acttttcttc atctcctgtc gcagccacga aattttccct ccccagttca ttcctgagtc 18540 aataagacag gacgcagaag ataacgggga aggaacacct ggacccatgc tgagagtaag 18600 aaacctccga cagggtcaac tacaaaagat cgtcgatgaa gtcaacagct atctcccaga 18660 tgaatcccat gtcgctatat ctctctttaa tggcccccag aacctcgttg ttaccgggtc 18720 gcctctcagt ttgtacggct tgagccgcca cttgcgtgga ttaaagcccc caaatactag 18780 ccgcgaagca cagatcccgg ctagcaagcg aaagcctaag atcgagcatc gcttccttcc 18840 cattacagca cccttccaca gcatatacct cgatagagtc gtttctctag ttatgaaaga 18900 cctcggggat atcaacatca caggcaacca actacgcatt ccgctttacg ctacaaccga 18960 tggccacgac atgcgttctc acggagccag taacctaatc ggtgatatcg tgagcatgat 19020 tgcgcgcgaa ccagtacact gggagaatgc aaccatcctt ccgaacgcta ctcacataat 19080 cgacttcggt ccgggtggca gtgctggtgt gggcgctgtt actatgcata acaaggcagg 19140 ccagggtgtg cgactgatcc taggtggctc catgaagcac agcgaggagt atgggaataa 19200 agctgaactt ttcaaccgtg actgcggcta tcctgttgtg tatgcggaga actgggcgga 19260 ggtgtacagg ccgcgcctag tcagagatgc agcgggtaag gtcattgttc agaccaagtt 19320 tagcgagttg ttgggtctgc ctcctattat ggttgctggt atgactccca ctactgtgcc 19380 atgggacttt gtctctgcta ccatgaacgc gggctaccat atcgagcttg ctggcggtgg attackacagt aagggtgcct tggaggaggc aatcaacaag attgtgcaga acggaatccc agggcgaggg gtaacatgca atatcattta tgccagcccc aactctgttc gctggcaaat tcccctgctg caggagctcc gtgccaaggg tgttcccatt gatggaatta caattggagc aggtgtcccc tcagtcgacg tagccaatga gtacgtcgag acacttggac taaagcatat cgccttcaag cccgggtctg ttgaggctat tcagcaggtc atcgacatcg cccgcgagaa caagtccttc ccaattattc tccagtggac tggtggcaga ggcggcggcc accactctta tgaggacttc catgccccaa tcttggagat gtatggtcgc atcagagcgt gcaagaacat cgttcttgtc gctggtagcg gctttggtgg tgccgacgat actctgcctt atttgactgg 19920 cgagtggagc aaggcattca atgcgccttc cctgatgcca tttgacggta tcctactcgg tagtagtt atgacagcca aggaggccca taccagtatt gaggccaagc agctgattgt ggatatcccc ggtatagacg atgcggtgtg ggaccagact tacacacgac ctactggagg aatcattacc gttcggtccg aaatgggcga gccaattcat aagatcgcaa ccagggggagt 20160 actcttctgg gccgaaatgg accgaaccgt gttcagcatt gcagacaaaa aagcaagggc 20220 tgccttcttg caagagaaca agaagcgtat tatcgaccgt ctcaacaatg atttccagaa 20280 ggtgtggttt ggacgagact cagatggcaa ggcctgcgag ttgaacgaaa tgtccctatc 20340 cgcggttgca tggcgaatgg tcgagctact ctacgtcaag catcaggccc gctggataca 20400 tcacagcctg cagcagttga cgttcgactt tcttcaatgg ctcgaggacg tcgttggaaa 20460 ctcatccggt aacttcgagt cccgatactg gcagacagtt ggccagatcg aggagccctt 20520 ctcggccatt gaacagttcc tagccaaaca tgctagagca acttcggagc tgatggatgt 20580 accaagcaca cagatgttta ttcacctctg ccagcgtcgc ggccagaagc ctgttccatt 20640 tattcctgtt ctcgacgagg acttcgaggt atggtttaag aaagattcac tgtggcagtc 20700 ggaagacgtt caggcagtgc cgggccagga tgttggcagg acctgcattc tacatggacc 20760 tgtcgccgcc aagcacacca cggtaccaaa cgagccgatc aagcatgtgt tggatggtat 20880. ccctctcag tgggtcaggt cccttacagg gctgggacat gacaatgaac aagctcagac gccactgacc gaagagggga tgagtgaagt gccaagcga atatctctaa cccgaacata cgacccaatg cttttcggtg aggagctgcc ctcaattgag ccctggatgc aacgactggc tggcacgcag aagggctgga gacatgctct cttctcgcag gtttctatcg tccaaggga cagtctgtgc throwcccca ttagacgact ctttgctcct cgacata cctttgtcga smuggling tgcactgcta ccgaacagtc tgtcgttcgc cttttcgagc gaacagcaga tgtgctgcag atcttggcta ttgaggtgcg tgctgttagc gaggacgtaa tcaaggccac actgttcgaa aatcgcactg tcactggagc accggcgggg ctcgagcttc tatttaccta tcatcctgag tttccgttcg cgcctatccg cgaggtaatc tccgacaaga ttgagcgcgt 21360 tagcttt tatcatcaac tttggttcgg tgagccgctg agctctgcta gtgttctctc ccaggaagat gaattctacg gtgatgttgt tgcagtaacc gctgaggata tcaataaatt caatcagtgc atccgcagta cagccacatc aagatctcga tcgagctctc agcagacact 21540 ggatgctccc atggattacg cgatcgtcgt tgcctggaaa tcggtcatga agccgctgtt 21600 cactaagagc atcagcggcg atctcatgaa actggtccat ctttccaatc aatttcgcag 21660 acttggaagc tttgaaccca tcaaggaggg cgatgttctt tcgtcaagag cacatatcca 21720 agccatcctg aatgaagact caggcaaagt tgtggaagtt gccgccttga tcagcaggga 21780 tggtgtggat attgttgagg tcgttagccg cttcatgtac cgcggatcat atggcgacta 21840 tgacgcagcc tttcagcaga caaaggagcc caaggtcgag ctccagctca caagtctcaa 21900 ggatgtggtt gttcttcact ccaagccgtg gtttctgcct cgggacaaag atctagatct 21960 caagggttat aagctcacat tcgagctgga aacgcatgcc cagttccaaa gcaagtcggt 22020 tttccgccag cagaggtgct ttggtcacgt ttatggtcgc tccccagcag gcgagactct 22080 cgaggtagct tacattgatc acaccgtggg cgtctcgacc agcaatccca tcatggacta 22140 cctcaagcgt catggaagtg tagtagagca gccggtcctg ttccagtcac caatcacact 22200 cggcggcaga gagatccaat tcaagtcacc ctcatccaat caagactatg cttctgtctc 22260 cggtgactat aacccaatcc atgtgtccaa gacctttgcc gcctacgcag agcttccaga 22320 cacaatcacc cacggcatgc acatgagcgc aagggtccgt ggcttacttg agcagcttac 22380 tgtccctgga aacccaagtg gctttaagcg ctatcagtgt aattttgttg ggatggttct 22440 tccggaggac gatgtggaac tggtactgca gcacgttggc atggtgaacg gacggaagct 22500 tgttcatgca gaggcccatc gcgccggtag cggggacaaa ctgatcgttg cggaggcaga 22560 aattgagcag cccagtactg gtttcttatt taccggccaa ggaagccaag aaaagggaat 22620 gggcatggaa ctttacgcct ccagccctgc agcgcggaag gtgtgggatc tggctgacaa 22680 gcacttcctt gagaatttcg gtgagtatca tctctttcac atgagtgcat ttctaacgtt 22740 accaggcttc cgcattaccg acattgtccg caatgatccc agagaactca aaatccactt 22800 cggtggtgct cgcggacgcg ctatccggga aaactacatg gctcttcgct gtgagatgat 22860 ccatgcggat gggcgcacaa cctcagaaaa gatcttcaaa gacatcacac atgataccaa 22920 gtcacacaca tttcgatcaa gcaccggact tctctcttct actcagttta cccagccagc 22980 actgaccctc atggagaaag ctatcattga agacatgaaa gacaaggggc tgttggcaga 23040 ccactgtagc ttcgctggcc attctctggg agaatactct gcattggcag caattgcaga 23100 gattatgcct attgagagtt tggtatcagt ggttttctac cgcggtttga cgatgcagat 23160 ggctgttgag cgcgacgagc aaggaaggtc gaacttctct atgtgcgccg tcgacccgag 23220 tcggctgtcg aagtgtatgt ccctttactt cctttcgcga ctggttactg actacgtcta 23280 gctttcaacg aaaaacgcct gcagtacttg gtatcgctca ttgctagaga gacaagctgg 23340 ttgcttgaaa ttgtaaacta caatgtcgcc aactctcaat acgtctgcgc gggtgatgtt 23400 agtcaatcaa tttcactcct aaagaaactc ctactaatac atcctagctt cgagccctcg 23460 acaccctctc ctcattgatc ggcgaactca agcgccgtca atctgacttc caggctattc 23520 tccagaatac tcaacccgaa gaggagattc gtgccagcgt actcgataca ctccaccaat 23580 gcgccaacgc cgccaaagca aagccccaac ctatccagct tgagcgggga gtggctaccg 23640 tgccactccg gggaatcgac gtgcccttcc actcatcctt ccttagccca ggcattacag 23700 ccttccgtgc atgtctctac aagtacattg acaaggatac gctggatgcg gaccgcctgg 23760 tgggcaagta tatcccgaac ctgacggcgc ggccatttga agtcagcgaa ggatacttcc 23820 ggcaggtgtt ccggttgaca aagtcaccga tcattgcgaa ggtcttggca gaggtacgtt 23880 gtctatcttg aatctctagg cttttatgtg ctgatttggt tgtttacagt ggcgagagta 23940 tatggatggt gagcgtgggt tggaaaaggt cactcggtct tcggaggttg tcgttgcata 24000 gctgcatcag atttacatag ggttttggcg ttgttgatac cagaatgtct ttcatgataa 24060 gcaattctat gattatatat gacattctgt cctttttcta caacccgtga tctgtcagga 24120 gaattatcaa attgaagtta agtctaaacg gtgccatgct ataaggtatt tctctcaatt 24180 cctgcaacta tctggctaca ccaatgaaga tcacgaggta cattgaacag tccattaacg 24240 ctcagctttt tcacaaattt gaattatgtt cacgaagcat atgctcattt ctccttaagt 24300 tggctatgtc tatatgcctc gagatctaag aatattccac tcatgttctg caaagcccat 24360 gctacttttg actaagacca agctacccac gtctatcagt cacatcgacc tcacttcaga 24420 atacaccaat cgccttcccg tcccccctaa acatcgcatg caatccatcc aacgcaagga 24480 atgactttgc aacttccaaa cccctcttcc tctcctcctc gacctgctcc cacgaccaga 24540 acccctcctt catcttgacc cctttctccg gcaacaatcc agcccgcgag agcactgtct 24600 ccggactcac tgatactctc cccttcttga aaacatacct cgtgacagca gaagccaaaa 24660 tacacttctc atccttgaca cccgtcgcgc tgccccaaa cactacctcc cctttgtctt 24720 tcttcccgcc ggtcttgaca aaatgactaa cgagatacac ccatttctca tcccaactta 24780 gcactctcgt atggatctca tacgcttggt atggttttat ctctctctta aagacacatg 24840 ctgtggatcc gagtgcgata ttcagctctc cgccggagaa ccccagcgcg agctggtctt 24900 tgaataaact ggagatcaag tgcacgcggt tgatgtctag gtcggtgaag taggttgcgt tggacttgtg catattgtag tcaatttcga gtagcggggt ggaggatgtc gtaatgaccg ggtagaagag ggattgggggg ccatggggtgg ttatgaccac tttacgggca cggttgagga 25080. tgatgtgttc tttgaggcag ttgaggaggc ggacctgtgt gtttgattgt ttttttagta 25140 tgtcgaggtg ggttgattga gtaattgaat tacgaggcgt ctgaagtgag ggtaatggga aggggacga acgtgccaga caaaagggag gcccttcagg tttaggatga tgtaggcccc 25260 tcctgcgaag agcagggtgt tttgccaggt tattgagttg aggagggctg ctgtgtccat 25320. tttgccttga aaatggttgt ggtgtcaatg tgcgctgga aggatggatg gctttggtcg 25380. 25440. 25440. 25440. 25440. 25440. 25440. 25440. 25440. 25440 ctcaatcaat gactgtcgga accactgtca gcacggtgcc tcctattgtt acactcggtg ctcgagtgtt gcttgaccca ttccctgcag aggaagcct tgctagctct tgctgatacc 25560 atggtccagg tgcagcttca gctccctgca cctgcaggcc ggatccttcg gcgttttgct ttcttgggtg ggtaatacaa cgtacattat tgtatcgaga tacatgacat gcatacatgc 25680 ttgtttaata cattacatac cggtattaca aacataccaa acatacatac gccaaacctc 25740 caaggatgta tacagcacca tttggacacc cgcactcatc tcatatatac gcagcatacg 25800 atatatatca ttcgctgtta atgcccacgt gggaccaagc ctcccgacac gctgcggcag 25860 gatgcgcaga tccacctatg gagtcgcatg gcctgccatg tgccaggaac catcgtggtg 25920 gagatgggag accatttttc cccgagatgg tgtccgtgac atggttcatt taccacgttt 25980 ctcttagctc ctagctgacc gatgaatcat ttgggatgct catgacgagg ggcgttacac 26040 gtggtggctt tactggtgta tagacatggc ttgggcggga ggccatggtc agggggatc 26100 atcggtgttc gggtgttgta aaaacaatgg agggtgcctt ttcagtagct taggagcttg 26160 gtgatatttt ggtggattgc gtgtgcatag tgcctagaca ttctggtcgg ctcccatataat 26220 footattggt ctgaaatgtc ggctgttgat tatgcaatac tgagttcgcg tggcatcatt 26280 ctgaatgggc ggcgactgtg ccgtagggca ttgccatctt tgactcatat gggcatatga 26340 tgtggctcat gtataacact tgtcctattg accaggagac ttcaacttgt aactacgatt 26400 ggataatact catcgtacta atactaccaa gtacgtcgta gatgttactc tcagtaggac 26460 agtatcaaaa tatatatccg aacattgaac attcgatagt attcactcat gatggatatc 26520 caattccacg tgagtgagcg tcagagttac agaaagaaaa tatatagatttt attcatgaga 26580 tgcttttaac ataggacaca tcgtaatctt ggcattgtcg atgttggggt cgccatggtg 26640 gtgaggacat cagggtaggt aaatctcgat actgggtgaa aaatgtgtcc cctcattaag 26700 ctagaacg accgtccata tataaactgg gtcccatagc tcagttggtt agagcgtaac 26760 gctaatagcg ttaaagtcga gggttcgagc ccctctggga ccatcttttt tcctatccgc 26820 cgggaccggc aacttatttt tcgtggccca cctacctaac aaacagcaga ctttagatgt 26880 tgtgagcacg aaagaaagcc cctgcgtttg ttccatttct tgtttctctt tcgtcaatag 26940 gtcaaatagc acgaacgagg aacaccactt atgtctcgta ggtctgggac ggttaatcac 27000 ttatcatct tacatgaatg aagatgacag gcaagacagc cgtgaagacc caccgcggtt 27060 agacaaaaac aaaagaaagt ccttggagca caattaggta gcgaagtgta tgtagcttac 27120 cttcccctt catattgact taaaacaa tatgggcaaa tgcgtctaaa tatcgcgcat 27180 acagaatagt gtgcagtcaa attcacacaa gcaggaacga ttcagattta tcacaaacag 27240 gttttccaag aataagccaa gaataatttt ttctatgagc aatggttatc agcagaacat 27300 cagactatct gccacaagga gtggcctaaa acaatcatat atgaagggtg aatcagaatc 27360 atcccctaac gatccagaag aagggcgccg cagttagaag tccctccacc ttgtatcgtc 27420 ggaggtacca agcaacccgt cttctgtgca atcttcaaga tctctctccac caccacccga 27480 gcatccttca ttttgatggg aacccgatgg tttccctccc actctattaa agaagctgtc 27540 tcacgattac aatatctctc cagaagttcc cgatgatgtt ccaacccggg atcctgcagc 27600 ccgtgtacat gtatggtggg aagatagaga accgccgaag acaccgacgc catagcatcc 27660 atatcaagag gaatcatggg gcctctcccc gcaagcaaga cagcgaagcg gaagttgact 27720 ccaccataca ccccaaccat cccagatccc ccattctgtt cctgaaagag caggctagca 27780 gccagccttg cgccctgact gaaaccaagc actgcaaccc actccccatc tgcccctgat 27840 cggtcatcct cttctattgt atgacggagg gagttgagga ttttctttgc aacgttggta 27900 ggtgagatcg aggggttcgg cggcattggg atccagctct tgaagtcacc gaagtgtcga 27960 tagaccgaga cgacatctgg gcctggctgc gaggggaaag gcgcttcggc gaagcagagc 28020 cggaagtgag ggcttagtag tgtgctcagg actcggcatt gggcgcggaa aatgcgggcg 28080 ttggtgccgc ctccatggag gcagaggatg cgtggtaagc ggagtgttgt gtcgtctggt 28140 agtttggcgg gcatactgtg tactaggcta tgatttatcg agagtggaaa gttaaagaga 28200 tgatggattt ggtgatgaag ggagttatga acccgaggaa tggaggctca tgctacaaaa 28260 gcaggacagg gttctacata caaagacaca tgggtgccac ctgtcaggtt gggacagaag 28320 aactatcgat gtgccagacg tcttagattg atcgccagtg agatcttgct tttctttctg 28380 atacgagatg aagatcgcgt taatagctct tgattggtca tccgctactc tgccacgcgg 28440 agtttcaaat agcccttatt cggagttttg gctagtgcct cttacccaat gtactccacc 28500 atggaagtcc actccgaatc ttgattgaga gcctccatat cggcatcagt cagacaggtc 28560 aacgagtcac attctccctg agggttctgt ggaaagaagt atagcagact gggaaacgga 28620 atgccaggga aattaggtcg ccgaatgaag tccggtcgcc ccagcttccc aaacaccgga 28680 aacagatcta cgcgtagtac cgacgacgac cccacatctg tgtctgggtt gaagcgcccg 28740 ccatatgcaa tcgtggactt gtcaggctcc tgcgcgatga atgtcgcaaa gcttcgcacg 28800 tggtaagcgg tattcgcccg gttcaattcc cggcgcatgt gacgcgcaat ctcaccgagc 28860 ggcagatcaa ccagctctcg gaatgtgagc caagtggtga cattgtgaac tagatctccc 28920 atgtattcgg cagggatccc taggaccttg cggccgtcca tggcccggga gaatcgggac 28980 cgcgcgtccg gggtgttccg tcgacggtgt cgcaccgtta tgtacttctt ccaacaaaaa 29040 gcactgacag catcgtcggt cgtgatgtag ggcacctctg ggtctggggt catggtttga 29100 ctggcgaggt ccttcagctc ggccagcttt gctgcggtga agcgcaagaa atgccaccgg 29160 gcgggatcag ggcgtaccac tggctgcgcg tctgtgatcg ggggtcgaat gtggtgggaa 29220 tgatctagca tgggttcgtc cggggtcaat agcgggatga tgttccgtcg atcgcgattg 29280 acctgttcca ggagagaatc ggagaattgt tcaccccgca tagccatggc gaccaggcgg 29340 gcgaagccga aaagaccgcc tgcatcggtc atgttatgtt gggtgacaaa gtcgagaaag 29400 actccccccgt cgacaaagct ggcctggagg cggatcaccc ggaatgggtc ctcctccgag 29460 tcctcgaagc gctgaggaaa ggcatttgtc ggggctagca gacgcgagtc cagcatcgaa 29520 ttgggtgcct ttagacgttg gatctccttg taggaggga aggcgtgggt aacatcgcgc 29580 accacgatga gaggatccgg aggcggagacag ggaacaa tgtatcgccc tgaattgtct 29640 gggcctttgc cgacgttgac gactttgccg gccatccagg gtacatgagc tcggatctgg 29700 cgcactgcag cttctagatc ggcgattatg gtgtcttgtg aaatccttg gggcatggca 29760 aatggaagca tgatgtgact gtagacttgt agcatcggca gctgaccgag gatatcctca 29820 taccgaccga ggtcttcata ctctggtcgg aaaccgttcg ttgccatggc ttttactgat 29880 gagcttggct gctcggtgga agggcttgag ccggagggaa gtacatttag atatagacat 29940 tgatgcttgt tttcatgtat ttgtatgata ttatactata gccctactac ttattgtttt 30000 tactgccgga cattcaattc gatgaagaat aagtgagcat ctgatcgggc acacccggag 30060 tttctcgtgc ggacttgcga tgcccttggg tatccgcacg tgtgcggaga tctctgccag 30120 ctcatccttc tggactaatc agcccacgac gttttgtccg agccgaagat ggacgtttcc 30180 tcaccttgag cacagctctc tatttcaaca ccctggatag ttgatcctgc gctgttagag 30240 taagaaaaaa gaaaacaaga attcgagagc tcacgatggg gtcccttgca aagaaggttc 30300 tgcttacagg tcagtgcaat ccattgttac ccgatacccg cagctgatga actaatgtct 30360 gttgagtaat gtaggcggca caggcttcat tgcctcgcac atttgactg agctactgga 30420 cgccgggtaa gcaatcctcg cgcgagacag cagatccttc taatagaatt tcaggtatca 30480 ggtggtagtg accgtccgca cacacgataa gggccgccag cttctggaat cgttgtcgaa 30540 caccggcgac aaggcggcat cgtacgtggt ggtggaggat atcgccaaag atggggcgta 30600 tgacgaggta aacctcacat ctatgcaggc tttagcagat gtccagagtg ctgactgact 30660 gtgcaaaatt aggctattca atccatctca gacctggacc tcgtagttca cacggcctcg 30720 ccgtatcatt tcaactacac tgatcctaaa acagacttcc ttgacccagc gatcaaaggc 30780 acagccggac tgctcgcctc catcaaggct tacgcgccga cagtcaagcg cgtagtttta 30840 acttcctcat ccgctactat tgtcactcct cccaaccacc cggaggtata cgatgagacc 30900 agctacgggt cggtgacctg ggaagaggcc atggtgccgc aagttacata tcgcgccagt 30960 aaggtatgag ctacttccat ccctgcacta caacagaccc tcattgacac acgcgacccg 31020 gaagatattc gccgagcgcg ccgccttcga cttcatcgag aacgacaagc caaacttcga 31080 ccttgtcaca atcaaccccc cgctagtatt cggaccgaag ccgcgccatg ttactgatct 31140 caaagcacta aacacctcga accacatcat ccgtgacacg atgctcggca agtggaaaga 31200 tgggggcgca cctatcgcca tacctttcac ctgggtggat gtgcgtgatg ttgccttcgc 31260 tcaccgtcaa gcgttggagc tgcccgaagt cagcggccag cgctttttca ccgtggcagg 31320 ccatttctcg aacaagcgaa ttgcggaggc tatccgtgcc acgcacccgg agctggccga 31380 caggctgcct cctagtgatg tgcccgacga tttgccagag ggtatttatg gatttgataa 31440 tagcaagtct cgtaaggtgc tggggatgac ttcgcgac ttgaacactt gcgtgggcga 31500 tgcggtgacc tcaatgctag aacaggcttg agtgtagcta ttcaagataa gttttcgatg 31560 gcatacgggc caccaaatgt caagaagcat gataaggata gcttaggcaa gattgtgaaa 31620 gccggttctc aatgcaagga gatgatgatg tgatcctagg aagacatagc cgcgtagcag 31680 gcaagatatt agattgagac tgtctactgt agatgctcat aattcaactg cccattcatc 31740 tacaacttct caatcctttt gtcctccaac aaccgcttca gaatcttccc actcgcattc 31800 cgcggaatcg catcaataaa acgcactcca ccatccaacc ttttatactt cgccagccgc 31860 agcgccacaa actccttcac ctcctcctcc cgtagtgact gatcctgtct caccacatac 31920 gccctcggat actccgtctt cgcctcagca tcgtacaccc ccacaaccgc gacatcctga 31980 atgcgcggat gattcctcag cacgccctcc acctccgccg gcgcaacttg gaaccctcgc 32040 accttgataa tgtccttttt ccgatcaagc agataccact ttcttgtctt gccgtcgcag 32100 tacccgaggt cgcctgttct atagaacccg tctttatcga aggcgctctc attggcggaa 32160 gggagattga agtatccacg cacgatggta ggtccccgta gacagacttc acccgtagtg 32220 tccggggcag tgatatcgtc tccgtccgtg ttgattatct tcgcttccat acccggaagt 32280 aaccgcccta cacttcctgt gtcgtcggat tcagggtatg gtatcatggt cgctatacag 32340 gaggactctg tcatgcccca gacttgggtg aaggttgcgt cgggggcgag gagagattga 32400 aagcgcgcct gggttgtttt gtctagtggg gcggcgccgc agcggacgat tttgagggat 32460 tgtagtgagt gggagtattt cggggttgtg cagtgggaga ggattgtaag aataattggc 32520 gggacggctg tgcattcggt gatgttgtgt ttggcgacgg tggttagaaa aaggtctggc 32580 tcgaagcggc gaagcacata ggatgggtca ccaagacgga gggtgctgaa gtgtgcagat 32640 gggggctggg cggcgtggaa gaaggggaga gagaggatcc gacttgtcta caggacgtca 32700 gctagatgtg gggtactacc tgggcatgca gttctgacct cgaacgggac ggggtgtgca 32760 tcgtacacca gcgtatgttg cgcaataata ttgtagtggg taaagtctac tggtttaggg 32820 agacccgtag tgccggaact gaagagtctc gctactacgg tcgttttggc atggttggga 32880 tcatcgagac ggtgccagtc agcctcgcca tgtttcagca gggcggtcca atgagcgagt 32940 cccgtcgtcg ctacaacctg ggactcgcgc gttgtgaaag cccagatgcg gtctttcgga 33000 atgccgttgc cctcagcggc agggatcatg cgatctagca tctctggatc actgatcacg 33060 aatttggcct gcgctactcg gaggtgatag ttgagttcgt gggatgtata gcccggattg 33120 gtgcccatgt atattcctcc agctgcgatg atgcctagca caaggatgct gtaatagatc 33180 tggtcgatat gtaagcggag atacgtagga aaaataatt atctttggtt ggggtgcaaa 33240 catcattgaa agaatgaagg cacacgcaat cgccgtcctt cagaccggct gctcgcaac 33300 cagcggccaa ctggcgcacc agagtgcggg cttgtcgcca cgagatggaa cggctcgtat 33360 cggaggcatc catatagacc tgaggtgtt tgttgcataa gtgtccagct ctcattggt 33420 tgggatgat tgactcaccg gcttttcgat ctcatatcgt gcctggtcga atatccagga 33480 ataatatcc tggactggga tgtcaatgct gcctcgccc tggaaaaaca tgattaaaat 33540 atgagaagcg gggaaaata cgaagaaagg atctgtgtca tgtagtacga gaggggtaga 33600 cttcaaaggg ctcttggctg ggcctataag ccctagtcac ggtcatgtga attagtccga 33660 ggcttgagca tcgagaggcg cggctagcat tgaatgccga tccctcag gttcaaagtc 33720 cgatcgtaaa ctccgagtga agatcgcccc gccttacgc agcacacct gagcagagga 33780 gcctcacaaa aaatatgcct cgttattggt caagtgga tcgactgtat atacagcag 33840 tgacactaca gctccaagga ctcctccaat acggaacag ggttactcca gctgaatcgc 33900 tcttcaggat cccatttttg cttcagccga gtgagatttt ccagcttttc agcggtatac 33960 cacacatctg gcccctcatc gccatgagcg tagttaacat agacggccag attgtcgaat 34020 ccactggact gctgaaagtg ccggcgagac tgtttcagga aggcgttgac cgggtcttca 34080 aggtaggggt cggtatacca gccctcaaga ttgctgcatg ttaagagtta gtgtctacac 34140 aaattttggt ggctgaggag ctccttacat atgcatcttg atctgccggt ggggataggc 34200 cgtattggag tccagcgtgg acagggctcc ttgcgtggga tagcgttgca caaggaagcg 34260 gccatcgtag gtgggattct ggcgatagaa ctgcagtagt tgctccatgt aggactccca 34320 ggcaggtacg tcggtgtggt tcagcccgat gctgtatata ttgaccgctt ggttcttagc 34380 gcaggcccct ccttcctgag ctgctaggcc gaagaaggtg gtgtcgagca gggcattcca 34440 gggcacagtc acactgcggg acatgattgg ggtgagactg gtgaaggggc tcagtagctc 34500 cagcgcctgc tcctcagggc cgtagtatat agcgttgacc agcacaagtg gcttttgaaa 34560 tgatcagtac ccacaaggaa attcagttga tggatgctta cctggtcgat cgtccggtta 34620 taggccaccg ccagggtcaa tgctagctca ggcggaagcg tatcatcgaa cgcgctcagt 34680 gcctgccaaa tggaagcgtg ctcagtcgcc gcgaagatga agtctacatt agtgacaagg 34740 cccccatggg tggcttggtg tgtgcgatag gtagccgagg tgacgactcc aaagttggac 34800 ccagcaccac gcaggcccca gaaaagctct gggtactgtg tctcagaaac ctcgataagg 34860 tctccaatcg gggtgactag gcgaacggac tcgagcgaat ctagcagtaa cccatgcagc 34920 ccctgcagac ttccgatacc gcctcccagg gtcatgccca caactcctac gcaaggagct 34980 gacggtgtca ctatttcccg ttgtgctgag tacagaggat cgatgatgtc cgagaacacc 35040 gtcgctccgg ccacagtcag tcgctcagcg tccacatcga gatgaacatc cttgaactgt 35100 tgcaagtcga tggcaatacc gtggcggaac cgctcaaagg taactgtggc tccgtggcca 35160 gcaccgacta ccaggaaatt gatggcatgc tctcgtgaga ttttgatcta aactccactg 35220 cattatcagt tcatgtcgca caggcttggc gaggcatacc gacaatatgt tggatgtcat 35280 gctcggttgc gggtataatg gcaccgagat agtggggtgc gccatgcaca gtccaacgtt 35340 gtgtggtgac atgcgtgtag ttggcatcag atgctaagac aatttttgca ttaggtgata 35400 ggctggggccattackatccc tggaggtcaa gtgtagtctg ggaggcccag agccatgata 35460 gaagccaaga caggatgcct gacagctgca tggcgtaaaa gattctggat tgatatgacc 35520 tagcgcaagt tagagccaat agttcttagt tataatactg tccaatttta tcaaagaaaa 35580 ctacgggtct cctgcgtatc tgatcatcag tgcgataagc agatggcaaa aaccccgagg 35640 ggaaactccg gtagttggaa gggcactaat tcaaccccga atcctgacta tcacagctct 35700 ctcatactat aatgactagt agctgctgat acaacaaaca aaaacagata tctgaagtgt 35760 cttcgcccct gtccttgccg catacaccaa gtcatcagag caaaacctct aaccaggcgg 35820 tagcctctat atccactctc aacgtctaca aataggcaat aaatcaaact ccaatatcta 35880 aatcaattga ttgcatcaaa taaatgctcc cgttatcttg gcagtaagca ctaaaatctt 35940 atgtctgagt aacggccatc ctgcacccca cctcaaaaca atgaagagat agatagaccc 36000 cttccaatcg actcacacct caccaactca caatctacca tcatgagtac agactcgatc 36060 gaagttgcca ttataggcgc cgggatcacg ggaatcaccc tggccctggg cctcctgtct 36120 cgcggcattc ccgtccgcgt ctacgagcga gcccgcgact ttcacgaaat tggagccggt 36180 atcggtttca cccccaacgc cgaatgggcg atgaaagtcg tcgacccgcg cattcaagct 36240 gctttcaaac gcgtcgctac ccccaatgcc tccgactggt tccagtgggt ggacggattc 36300 aacgagtccg gtaccgaccc gcgcgagacc gaggaacagc tactcttcaa gatctacctc 36360 ggcgagcgtg gatttgaggg ctgccaccgt gccgacttcc taggtgagct ggcacgtcta 36420 ctaccggaag gtgtggtgac attccagaag gcgctggata ccgtggagcc tgcagcagat 36480 aatagcctcg gccagcttct tcgattccaa gatggcacga cagctaccgc ccacgcggtg 36540 atcggctgcg atggcattcg gtcgcgcgtt cgtcagatcc tcctaggtga agaccatccg 36600 acagcatcag cccattacag tcataaatat gcagcacgcg gccttattcc catggaccgc 36660 gcccgggagg cgctgggcga agataaagtg gcgacacgct tcatgcatct cggtccggat 36720 gcccatgccc tgaccttcc cgttagccat gggtccttgt tgaacgtcgt cgcttcgtc 36780 acggacccta acccttggcc atatgctgat cgctggacgg cgcaggggcc caagaaagac 36840 gtgacggctg ccttttccg ctttggtccg accatgcgca ccataattga cctcttgcct 36900 gatcctattg atcaatgggc cgtttttgat acatacgacc atcccccaaa tacgtattcc 36960 cggggagctg tctgtatagc aggggatgct gctcatgccg cggctccgca tcacggtgca 37020 ggtgcaggtt gtggtgtgga agacgcggct gtgctgtgcg ctgtgcttca tatggctgcg 37080 aaaaaagtta acaccgcaaa aactggttct gaggggaaag ccgctctttat cacggccgca 37140 ttcgaaacct atgattcggt ttgtcgcgag cgtgcgcagt ggctgggtgga aagtagtcgc 37200 gttatcggta atctgtatga gtggcaggat aaagggtag ggtcggatgc ttccaggtgc 37260 cacgatgagg tgtattggcg ctctcatcgc atttggact atgatattga tgcgatgatg 37320 agagagacag ctgaggtgtt tgaggcgcag gtagctgggg tggcgagaaa ttagtatacc 37380 tttgactact tattacaata tgaggtctgt agtgctagga atattctttc gtgtatgtaa 37440 aatttattcc accgcctatt gctgaactca tggggaccga gtctatgtat ctgcaggtga 37500 ctccaacccc cttgcaccat atagctgtta tttcccggct accctcttga gcatgtcggc 37560 tcaggcttcg actagcaagc ggtgtcctct tggagccatg atcaaatact gttcctgagc 37620 cagatcctca tcccgcatgc cgttctggcg cgacatctga acctctggaa accggcgtag 37680 gaatagagtt gtcgtaagtc ggatttccat gagcgcaaga ctattgccga tgcacactaa 37740 taacggtaac aattagtgat atttacccta tgatattgtg attaggctgt ggaagtcaat 37800 cttactcctc gaggcgccac caaagggcat gtaagcgtct ttcatctcct gggtcggaga 37860 ttcccagcga ctggggtcga atctacattc aattagccac acagacagca tcgaaagata 37920 tgaatacttg gggttgggtt tgtacttctc cgggtttgga aagatggcct catttcgatg 37980 taatgtatac gcttgagtcg aaaccgtagt tccaccaggc aggtaatatc cagcaagcgt 38040 agccccgcca gcaggcacca cacgaggcaa agcagacgag acaacagcat atttacgtag 38100 agttctgtg atgacctgat tgaggtacgg cagatgtcga agctcatcgt ggcggaaatt 38160 ctcaggcaaa gtacggattt ctgccagcag cttctgccgg atctcattgt ccctacagat 38220 cgcatagatg agatataata acgtgacggc agttgtatgg ctccctgcgg tatatatga 38280 gccagccaag tatgtcagct ggaagtcgtc aaacttgtct gccttaaaga gtttagtaaa 38340 taaggtaggc ttcggactct ctggctgttc ctggatatgc tgtctatatt gctgaataga 38400 ccgcctcgcc catttaccaa gaagccctat tgcctgaatg gcatgcttga aaggatgaac 38460 gggcatgaat cgtcccagta cgaagacaaa ggggaaggtc gtccgccagg cgtgcatgat 38520 acccactttg tccaggtagt ccatgaccca ggcggttttcc tgtcaaggcg tgtcaagatg 38580 catagtcaag tctctaatac caaagtccgc cactcacctt gggacgcgcc agcaggccaa 38640 tgggttcgcc aaagctcaat tcgctcacga cgtcggtcgc catatacgtc caccatttgt 38700 agaggtctga taccccctgt gactgcaaat cggaggcaat ccgatcaatg gtagcatgta 38760 catggttggt gaccatgggc tcaactttgc ccaagctggt atcagacaat ggcccggcta 38820 gtaatcgccg ccattcagag tactgagtgg gctcgatgag ggtgagcaag ctagtcaccc 38880 cgggtggtgt caaagacttg taccactcgg acttgagata tccgccgcgc acgcgatgga 38940 tctcacggac tgctggcaga tcacagatgt cgacttctgt aggagtcacg cggacaatag 39000 ggcctaccgc caggagcgcc tcataacatt agctataagc ttagtaatga tgaaagacgg 39060 gtatcgctca ccgtatttac ggtgtagata gtcgacatac tcaacctttg tccccgcaaa 39120 ccagtagtat tgctcgatca atgaactcca tttcgaaatt tgaggaccag ggatatgtga 39180 tagaggattc cttactgccc gatacacgat tgtctatagg ggctgagtca gctctaccaa 39240 ttgtggtgtt tgagatgagg gttgcctact gagagactga gagctaagac acagccagcc 39300 agaggcaaga aaatgcccgg tagctgggca aggctttcca tgtcgcagta acctacagct 39360 atggagatct tctcaaacat aggtgtcaag aaacaggata ctgaatttca agagccgtgg 39420 ctatctgcct gtctggcagc tgtagaactg aaggggcgtt gatatggtgt tttgcagctt 39480 tgatgactgg cagtagaaat tccatacaat aacggaatga tggattagag atagaaacga 39540 ctcgcaaaaa ctgttcattg gccacgaaat tgcaaagata atgagcaaag gcagctgcgg 39600 gagaaactcc attgatataa gtccgagcga cgcggggtca gaaaatcccc tagccacata 39660 ctcggacttc ggtggatcac agaatttcca cccctggaag aggaagcata cgatttgtgc 39720 aagctgatga gtactatcaa ggctttctcg taagagctat cttgaggaaa cgtgaagagt 39780 cctgcccggg cgaaactccg gctcaacagg atgcctcaca tcagcgagat aatgcctgca 39840 gagaataaga aaaagtccga gaggctacca tgtgtttctg ctgcagcatt ttttacgtgg 39900 cagtgaattg ttctggatct atacttgttt tgatattgca ttacaagtca tgcatttatc 39960 tggccatcac agcatcggaa aagcgccggt agggattgat tgattgcttg attackgtt 40020 cttcgacctt gcaattattc cttacatctc ggcttgccct tcagccttat ccgccatggt 40080 cacaacgaca agggctacta acccgacaaa tacccttctt ctattcggcc cgcaagccct 40140 ttccttctcc accgcgacgt ttgccgatat acatgcacga gtagtgcaaa cctcagagaa 40200 tgcttggatt aagcagacca tcaccagtct accagggctg tgggatgccc ttgtgaagga 40260 atttccacag tatggagcgc tggaaggcaa gcagttgctg cgggatctcg ataggtggtt 40320 cgagacgggg acgatggagc atgctgaacc ccatcttcct aacatccttc tcagccctat 40380 ggtagtcata actcagttga cagaatatgt ggactacttg aagacgatgc cgcatgcagc 40440 agatcagcaa acagaaaccg ttggattttg taccggcctt ttgactgctc ttgccgcatc 40500 acttgcccct gacatcaagg gtatccgcca gtacggcgcc attgcgatca agctggcgat 40560 gattattgga ggggtggtag atgtgcaaga tatcacaagc ccaaatggcc cttcaaaatc 40620 attagccgtg gcttgggatt cggctgaaac acaggaccga ctgaaccaga ttattgacca 40680 gtctccagag gtgagtgcac acagcaagac acaatgaaaa aagacaatct aattctaatc 40740 taaacataaa ggcgtatata tcggtagaat acgaccataa ccgaccgaca ataacaacag 40800 cagccagaag catctcttcc ttgcaacagc ggttacgcaa tgccggcctc attgctagcc 40860 agattggatt gcgcggtcgc ttccactgtg catgctatga gaagatatt gaggccttga 40920 gcaaattctg cgattctgta cccagcctgt gtctgccaga tgccgctgtg ctggctctgc 40980 cgacacgttc caatgatgcc gggtctttta ttctcagcgg gaagctccat cattgcgcac 41040 ttcgatccat tctcctggat acatcccatt ggtatcagac tctagaggta gtccgacaat 41100 cattcttaaa atcgcttcc tcgatggtgg tttcttttgg acccgagcga tgcattccac 41160 cgtctatact gaaggcctt agctcgatag ttaccacagc ggctgaatat caaccatcat 41220 acttgcacag agatccagag ctttgcaatc cgaatgagat tgctgtgatt ggcatgtcct 41280 gcaaggtggc tggtgccgac gatgtggatg agttctggga ccttctatgc aaggccgagt 41340 ctcagcatca agaggtgccc aaggagcgat ttggattcga aagtgccttt cgcgaggtcg 41400 accccacacg gaagtggtat ggtaacttca tcaacgaaca tgactgcttc gaccacaaat 41460 tctttaagaa gagcgcccgc gagattgccg caaccgatcc gcagcagagg cagatgctgc 41520 aagttgctta tcaggcagtg gagcagtcgg ggtactttac gacgcccaag tcagacaagg 41580 accgaaagat cgggtgttac attggggtgt gtgcagcgga ttatgagtac aatgtggcat 41640 gccatcctcc gaatgctttc atggccacgg gcaacctgaa gagttttgtg gcaggcaaga 41700 tcagccactg gtttggctgg actggaccag gactctgcat cgacacggca tgctcatctt 41760 ccttggtggc tgttcatcag gcgtgtcaag ctatcttgac cggcgactgc accgcagcat 41820 tagctggcgg tgccaacatc atcacccacc cgttgtggta ccagaacctg gctgctgcct 41880 ccttcctcag cccgacaggt caatgtaaac cgttcgatgc cagcgcggat ggttattgca 41940 gaggcgaggg cttcgccgcg gtattcctca agaagatgtc ggccgccatc gccgatggag 42000 acatgatcat cggcagtatt aaagccactg cagtcaacca gaatcagaac tgcaccccag 42060 tcttcgtacc caatgcgcct acactatcgg atctcttccg tgacgtgttg gaccgatccc 42120 agctgacagc taaccagatc acggtagtgg aggcccatgg cacgggaacc caggtcggcg 42180 atccagccga gtatgaaagc attcggaatg tgttgggcgg tccgtcacgg tcaacgccac 42240 ttcttttcgg atccgtaaag ggcctggtcg gtcatacgga gtgtacatcc ggagctgtct 42300 cgctggtgaa aacactcctg atgcagcagc atgaggcaat cccacctcag cccagctttg 42360 acagattgaa tccggaaatc cctgtgtccg agtcggacaa tatgcagatt gccaccaggt 42420 tttcaccctg gactgcagaa taccgtgctg ctctcatcaa caactacggg gcctgtggat 42480 ctaatgcgtc aatggtcgta gcccaggcgc caagaacgga gcagaagcgg agtgctacaa 42540 gacggacctc cgttgtcctc gactacccct tccgtctctg tggatccgat gatcgggcct 42600 tgcgagccta cagtgagcga ttagtgcgct tcatagctag tggcataaag gacggaattt 42660 ccgtggcaga tctcgctttc aacgtttgtc ggcagtccaa ccctactctg gatcgatcat 42720 tggcgtttgc ctgtcgtaca acacaggagg tggaggaaaa gctgagggct ttcgtcgccg 42780 ggaaccaagg actcatctca gccagtcggt cgaagacacc tcgcgaggtc atcttgtgtt 42840 tcggaggcca gatctctaat tatgtcggcc tggatcgaga agtctacgat aatgttgctc 42900 tactccggaa gcacctcgcc atctgtgacg ctgcatgtcg ccatctcggg gttgacagta 42960 tctttcccgg catcttccaa aaatcaccaa tttccgaccc ggtgaagctt caaacgattc 43020 tcttctccac ccaatactcc tcggcgaaag cgtggatgga ttccggggtc cgccctgttg 43080 ctgcggtggg ccatagcttc ggtgagctaa ccgcactttg cgtcacgggt atactgagcc 43140 ttgccgacgc aatgaagatg attgttggac gtgccaccgt tattcgtgat ttctggggag 43200 aagacaaggg atccatgatt gcagttgagg cagacgaaaa cagagtccag cggctgcttg 43260 ctgaagcagc caaacagtgc gagctgattc acgtaagagc acctaccatt gcatgcgtta 43320 acggaccgac cagttacaca ttagcgggac cggttaaaag cattgacatc gtcacggaag 43380 tcataagtag attatctgac tctggaccct caatccgctc gaagcgatta aaggtgacca 43440 acgcattcca ttccaccctc gtcgagcctc tgatggagga gctcgagaag gtcggacagc 43500 aattgacgtt caacgcacca accatacaac ttgaacgagc tattgagcac cactcggatg 43560 ctacgcttac aagcgactat gttcccgacc atatgcgcaa cccggtatat ttcaaccagg 43620 cagttcagcg gctggctcag cagtacccgg atagcgtgtg gctggaggcc gggtccaact 43680 ccaccatcac ttcgatggct agccgagccc taggatctcc caagtccctt cacttccaag 43740 ccgtgaacat cacctcggac gattcgtggt ccatgctgat tacgagtaca ctgagtcttt 43800 ggaaacaagg catatccact aacttctggg cctaccatgc gaagcagaca tacgagtaca 43860 atcctgtttt gctcccaccg taccagttcg agccatccag acattggatg gagctcaagg 43920 ttccaagctt catgagcaat gggaatgtcc aatgtggccc acgggatgaa gaagagcctc 43980 ctaaaacgct atggagctta atcgaggcaa gcgataaagt ggcccgtttc cagatcaata 44040 cggcggctcc gaagtatgtg gagctcgtct ccggtcatgt catcgccaac acggcgccta 44100 tctgtcctgc aactgtggaa gttgatattg tggtggaagc attgcgtagt ctgcgtccgg 44160 atttcatgga ctctaatcta cagccacagg tgctggcggt cacgaatcaa tcgcccatct 44220 gtatcgatcc gaatcgctcc gtgtggttgg aatgccaagc gatggactcc aacttagtat 44280 gggagtggag gattgtcagt gacagcttgc aagagcccgg aacatcctcc tcagctcacg 44340 tgctcggaaa gttggctttc ctttcagggc aagatgaggt gaaacaacaa gaaagtgagt 44400 tcatgcggtt ggagcgattg atcgggcacc aacggtgtgt ggacctgctc aacactaccg 44460 aggcagacga tatcattcaa ggtcgtaaca tatacacgac cttcgctggt gttgttgatt 44520 acggagagca gtatcgcgga ctgaagaaga ttgttggcaa aggtcttgag tctgccgggc 44580 gcgtacagaa gaaaccatcg gaagagtcat ggctagacgc acacctcgga gattgcttca 44640 gtcaggttgg cggcatctgg gttaactgca tgacggacca caatccagat gacatgttca 44700 ttgcgactgg gtttgagaag tgggtacgtt cccctgcgct gcgacatggc cagccaagac 44760 ctgagatatg ggatgttctg gcttgccatc accggtcgtc ggaacagacc tatttgacgg 44820 acatcttcat attcgacgct gaacaaggtg ctctgacgga agtcattttg ggcatcaact 44880 accacaaggt tgcaaaggcc tcgatgagta agattctctc gcgtctttct ggaactgagg 44940 cagccccttc gtcaagcaca cgtgcccacc caacatcatc ttcgtccccc cgactgcccg 45000 ggccattcgt gccagaggat aagagtcaga atgagacaca aactgcaggc acaaatgccg 45060 tcgcaaagaa gaagtcagag aagtcggccc aacagaacgt gctcgacaag acgcgagccc 45120 tcctggccga aatatcgggg ctagagccta gtgagatcga agctgagact ggtctggctg 45180 acatcgggat cgattcgttg atgggtatgg agctggcacg agatcttgag gcgctcttta 45240 aatgcccctt actcggtgat gagctggcga atgtgaccac cttccagggt ctggttgagt 45300 acgtgcaatc tgcggtcggg gtgccagcca acggcgacga acccgacaat actaatgcgg 45360 acgaagtctt cgaagaagac aacctcgccg caagtccctc gagcagcagc tcttccacca 45420 atcttacgga agacagctcg ctcgacccaa cagagactac aactaacatc tcttcctacc 45480 ctggtcagac aaaaacagaa aagcctgcca tgccgcccgc ttcatccaag actctagagc 45540 tttcccctgc gtgggtcctc gaagccttcg aggagtcgaa gcagctcacc gatcacttca 45600 tcgaacagta tcggtgtgct aactacgtcg ataccatctt gcccaagcaa acccagttgt 45660 gtgttgccct gaccgtggaa gcctttgaga aactggggtg tccaatacgc agcgccgtcg 45720 ccggccagaa gctcgagcga atcctccaca ttccgaagca tgcgcaattg gcacagtatc 45780 tataccgact actaagcgct gacgcacggc tcataaatct gaccgaagac ggtcgcatca 45840 cccgcacgca catggctctt cctaagccaa gtgatcagat cctccaggac ctcctccggc 45900 tatatcccga ccacgaatg gccaaccgtc tcgccgcttt cacgggcgct cgactcgcag 45960 aagtgctgaa gggagagaca gatggcctgg gattgatctt tggcaccgac gaaggtcgcg 46020 aactcgtagc gggcctatat ggagactctc tgctgaacaa gctttcatac cgacagatgg 46080 aggacattat tacacgactc gcctctagga ttcccagaga cagcggtcca ctgaagattc 46140 ttgaaatggg agcaggcacc ggaggtacca cgaggggaat ggctcctcta ttagctcgac 46200 taggaatccc cgtggagtat accttcaccg acctgtccgg atcgtttgtt gccgcagccc 46260 ggaagaagta ccagaaggaa tatccgttca tgaaattcca ggtgcacgat atcgaaaagc 46320 cccccagcga tcagctccga cactcgcagc acattgtgat cgccagtaac gccatccacg 46380 caacacacag cttgactgac tcctcccgcc atgttcgcga atttctgaag accgacggct 46440 tcctcatgat cgtggagatg acccagccgg ttcactgggt ggacattatc ttcggattgt 46500 ttgatggctg gtggctcttc gccgacggtc gggatcatgc gattgcctcc gcaggctggt 46560 gggagaaggt ctttcagtca gtgggatatg gccaagtaga ctggacagat ggccatcgcc 46620 cagaggtgca gattcaacgc gtgataatcg cccttgcctc agggcctcgc tatggccggc 46680 agcctctccc gccggcacct cccccgaatc tcgttccagg aagccatgct agtcgacagg 46740 ccgcggtaaa tgagtatctc gataagtata cgaaagggtt cacgctgccc gctcaaacct 46800 ctaacacaga catctccaac tcaaccagtt attgggagaa gcaatgtgtg ctaatcaccg 46860 gcgcgaccgg cagcttaggg gtgcatctcg tggccgccgt agcagcattg gacgacgtcc 46920 aaaccgtgat atgcctaaac cggcgcagcc caatggaccc agaccttcgc caacaacagg 46980 cattcgagcg acgcggtatc ctcctcgaag ccgccagcat gtccaagatc cgcgtcctac 47040 aaaccgacag cagcaagccg caactaggac tcaccgatga agtctacagc agcctcgtaa 47100 cctccacaac gcacatcatc cacaacgcat ggcccatgac cggtaaacga cccctatcgg 47160 gcctggaaca acaattccta gtcatgcgaa acctcctcga tctagcagca caatgctcct 47220 ctactcgacc agccaacgcc ccgcggatcg tcttccaatt catctccagc atcgcaacag 47280 tgggatatta ccctctatgg agtggacaga ccctggtgcc tgaggcccgc atggggatcg 47340 agtccgtact ggccaacggc tacggcgaag ccaagtacgt ctgcgagcag atgctcgacc 47400 gcactctgca tcaatatccc gaccggttcc gcgctatggc tgtccgactg ggtcagattg 47460 cgggatcccg gacaagcggg tactggaacc cgatggagca cctgtccttc ttgttcaaat 47520 cggcgcagac attgcaagtg ttcccagact tcacaggtga tttgtgctgg acgcccgtta 47580 atgacgtcgc tgctacgctg agtgaccttc tacttcgatc tacgcattca aatagtacga 47640 ccgaccaacc aatataccac atcgataatc cagtccgaca atcctggagc gagatggtac 47700 cggtgttgat cgatgcgctc ggcatcccgg ctcaaaatgt tattccattt gcggactggg 47760 tctgtcgggt gagggcattc cccggtcaag tggaatggga taacccttct gcattgttga 47820 ttgatttttt ggatgatcat ttcttgcgca tgtcgtgtgg agggctgttg ttggatacga 47880 agcgagcgtg tgagcattcg ccaactttgg cggctgttgg gccggtcact gccgagttag 47940 cgaggaagta tattcagtca tggaaggaga tgggattttt gaatccttga gtctaagatt 48000 tggttgattt gtttggtagt tgctagataa tttggacaat gaactgttgc ttagtgattg 48060 tggactatca cgcagagtct agattacgta ggctagagag aacgataatt tgcagaagtc 48120 gctgctatgg atagctgaat tggcgctcct gatggctgat gatatgtatt gcatttcatt 48180 tagctttagt attcaaattg attagatagg actacacgag ggtaacttgc tagtcttgtg 48240 taccacaagc atggcaatat atatactacc aatctgtcta ggcttaaatg ccgctgagtc 48300 gttgcctaat atactaccat cacaatggat taggagaacc ggacactata aatagaaacc 48360 atagtcaat atctaaaaag ttacaaccat gttcaactgg catcttatca tctaatctag 48420 tgagcagagg accaaaacgc gcgtacgacc atgacagcct cccgggccgc ctcgagcttc 48480 gagaccagga tcatgaccgg cggatgtcct gagctgctgc ttcctgtcga gccaatcgat 48540 tcacagtcat caatattc aaggcgtcct gcacctcccc taacagctt ggggtatcat 48600 tatgggcaac agggtgctgc ataccctg cacataggcc tagtagaata ggagtgaagg 48660 gatgcacctt gaagcagctg aaatggcgta gattacgtgc tagacgaca gtctcgcggg 48720 ccgcctggag cgcgcatgcc tcgtagtcgg gtaggaggct gactgtttcg ggctcccagg 48780 gcgtggactg gaggatgtca tacaggga ggacacccgc ttgccgatc atcttggcaa 48840 acaggagcat agggtcgctg gattccgcgg ccgtgggaag cgaaagctt agctgcaggt 48900 gggctgtgag cgtcgtgtcg atccactgat ggcgagtcca gatacctga agcggatttc 48960 ctcggaggag ctcagccatg gatagatggc ggtgtgacag agtgcggcca ctgatagtgg 49020 ctagcaggat acatttagca aaggatta ggggactggt atatccgta cccgcgagtg 49080 cttcggccaa aaagcccatt gaaatagggt gaccacattg gaattcactc tcgggcatgg 49140 ggagtcgggt cagcgcttgc tcggtcaggg tgagagagcc cttatggcgt aaatttaggg 49200 aacagtccat tatgtaggcc atccacacgg tgcgtcgttt ctcctcggcc accacccatt 49260 tagaatctgg gtctaggtcc aggtcgagca gcgaaagagg cgaagcgtct atctccggca 49320 ggcgcagcag ctgcactagt cggaatacac gaccggcgct catccagccg cgagagtatg 49380 agacctgcat aaactcgtaa atggcgatga ggatccacgc ttgcacctgc tcgaaatccg 49440 gggctgtcaa caaagtgtcg tccgactcga gggactccag tgtgtggcgc gtggtccgat 49500 acagtgctac gcgtacctcg ggcaactggc tactcacgcc agcggccagc gtgcgaatag 49560 cctgctgcag cccccgtcgg tttgccgatt gctgaggttc ccgtgcccag gaaaagtatc 49620 gatggcgttg caatatgggg accatcggct ggacgcgctc gaaaaacacc tggtccctga 49680 gccggtgagt ctgatgactt caatctcgta atggaaggga agataaagca tacaggtctt 49740 ccagaatgtt ttcggatact tcctcagagg ggtccgggga ctgggaagct ggcgaagagg 49800 cggacagcgc tttatccagc agctcatcgt ccgagatggg atcaacgagc atgccatcgc 49860 gctgctccag aagacaccgc tcgagggtgg ctgttcccag vakacataat cagcacaag 49920 ctcaatccctc aggaaggaaa ttcaggccc taccgatgcg gcctcag gcctttagat 49980 gaccctctt tggaccacgc gccgggcgtg caagttggt gacataa acaccat 50040 caacgcagtt ctgacactgc ggctggcggt cgcaccgtag cttccggcga cggcattcct 50100 cacaggccga accaggggccc tggcgcgagg tcttgtggtt tgtagtgttg ttttggtgg 50160 ttgtggttcg ggagtctgac atgatcagtc accgtcacag ttacgatcac aatcacgatc 50220 catcaagcgc gataggctga cttcttcagg gaagaagg cccttctt gataccagc 50280 gtacttacc aactcaagct gctgcctcat ggccacaaaa ggctcgccga tccagtgtgc 50340 aggaggccaa tggcatgcca aacacggcac gtcccacacc aagcgccacc aaatcgtcgt 50400 ttatctcccc ctcatcatga gtctctcatg gctgaccgat aaatttcaca gcccccaga 50460 aggattgaag gattgaagga tgaaaggcca taccacatcc acttgcggct tccaaaaggg 50520 cggcgtgttt gccatcggct cacctcatac ttccccatct tttcccttcc cgtgcgtcag 50580 cagcgtgcag gaatggattt gtgtatatac gaggcgaccg gggggagatt cctggtagtg 50640 gctgaccaaa gagaatcccc gagacgaaca taagcccgcc gataagcaga gccttgttgg 50700 ctccccaatc aacatcgatg gccggcagag gaatgctgaa acggccaaca tagtattgcg 50760 aagaatatag gattgtctct tatcagcgct ggaagggtct gatcactcgc tgccatatgg 50820 cagagtgcag tatgcaggat aggtgccgga gatcctgctg ctcgttcagc tcgcctaccc 50880 tgcgttctgt cggcccagac catgtggaag gacgggaagc acggtgcgtg cacagacgcc 50940 ggcgggtttt gctcatagta tggggccaat cgcagggcgc tggaagagag gagcaggtcc 51000 accataggcc gaaatgaagc tttgtggctc ttccgggccg ccaggttgat ttacttcttc 51060 aacaccccat atgagcttgg cctgcgaggc tggaagcgat tggatcatct agtcaactaa 51120 cgaatcctgc tgttgccacc cccatttccc ctcattgatc agtggaacgt gggagaaaac 51180 tccggtggaa agtccgggta attaagatga ctgaccccgc agtatcattt ttttttctga 51240 ctagtactac tacggaatat aatcgaaggc atcgataaag aataatcaca ccaatcaacg 51300 agccctgcac atcatgattc atcactcctg aaagccgatg tctcatagcc ggcttacggc 51360 aacgtcacca ccagcttctt agccgaaacc ccttgtttca aggcattcaa ggccccttga 51420 accgcctcca atccagtgcc caccacctca gcctcgggcg cagcaatgaa gcttccgtgt 51480 gctagagctt caccgaggta tccgtagagc tgatggctga cctcattatc cttgacggtg 51540 ctggcgaaga tgaacttgca ggacactccg tccggcacgt tcgggggtgc aggcatcgac 51600 aaagcgacaa acttgtcacc ctcggactcg gcaaccaggt taagacaagc ctccgccgct 51660 tcattgcggg tctgaggaac cacgccagca atagctaggg ctcctgcgca cgtcttgcct 51720 cgaaaggcct ctcgaagatc gtcacggatt gacggtttct tgtagtcgaa caccgccgta 51780 gcccctagac tcttgaccaa gtcgaaatta tgcggggaac aagtagtgat gacctcgtat 51840 cagccgcga cagccagctg gatggcgttg cagcccac tggtggctcc acccaatc 51900 agtagggtct tccccgtccg ggttggggtc agcgacggga tctgcagacc cagatagtcc 51960 ttttggaata gaccaacgat cgcggtggag aaggccagag ggacacagc tgcctgttga 52020 tattcgagag cgtggggcag aggaagcc atgttctcta gcacaccac atactgttgg 52080 aaggctcctt cagcgctgtt gttgctgttg gttcccactg cgtggccgac cacgcgatca 52140 ccaggcttaa agcgggtgac tcccggacca atcgcaacga cttcacccgc gatatcctct 52200 cccagcacca gtggataagt gacccacgga agagagtct cgcccatcat ctgagtcgca 52260 tagtcaccg cattgaccgc cagggcatgg acccgcacga cgatgcggtt ctctcccgct 52320 gttgggtacg gcgcttcctg gacgaccagc gactggttct tgccgggaac gatgcggcg 52380 aagttggtgg gcatattgat ggttgagatt tgaaagata gatacaac aaaattgatg 52440 tcagatatgc caagagta ttaagta agacggata actgcttatt tacggctgat 52500 ggggcagtgg gggaatcgtc tggcaagagg ggcagtgaac tatacaaa gcagatcggg 52560. tttggtgcaa tcccgacggc caggagata gataggctct cttaactccg aaagcttgcg catatttatg cccttccttt caatttgtcg gatgatcaac cttcttgcgt aatgtttctc aggggcgttc caaactcccc gtcgggattc aatgccggcg gagtttctgt tattgacggc 52740. catgaatgtg ttggtttatt tgtttcccct catggaggg cgatgaagta catctttctg ttcagatcta cacaggatca gcggccgtct gtaacgatta cgttgaacta ccagaaagga gagctgtctc tcgatagctt tgcgagcgcg gctagtcatt cgaccatggc ccctctgcga 52920. cccaccgatg ctgacgccag gaatgcatct tccgataccc ccgaaaagga tttaccatca gcaggtacca gcagtcgtg gtggccgtgg tcgagaacag ccgcgccttc tcgcgcccct 53040 agaggtgtgt gattacggcg atggtctagt cttcctcgct gatttaggca gctgagatgt 53100 attggccagc tgtgttaatt cggaggacga tgagtatcat ctaaagtcc catcgaaggg caaaagcaaa gagagatatg aggacttttg gaacgacgtt gtgaattttt tccatccatt ttctcgttcc aatggtgagt taaccctaca tagcatacag gtagatggag tatatattgc 53280 tatttgagcc acatgctgcc tacgtagaac aagacaaatg tgtttgtcat acaggatgga 53340 aaatatattg tatcatcatt aggaagcatt cgcaccactg tcttcttttt ttccatctag 53400 gaattcccgt atagcagcaa caatgacctc gtggtcggca gcctcctcct gcagcccact 53460 cacagcaatt acccccacga ccccttcgac tccctccacg cgaatcggaa acccacccgc 53520 ctgcaaggca tactcgtatc gttccaccgg tgacagagca aactgctcag ccaacacctc 53580 tatcccctcc ataactgaag gcccgacaca cacggactca tttgcacgag ttggcttcaa 53640 ccggcagtgg aggtaccagg tagacacctg ccagcggagg actgtcttcg tcttgcgctt 53700 aagccaaaga acatggtcca ctcgggttcc cggctctgta gcacagtgga aactaactg 53760 cggagctgat gagccagaca ggcagatcga gatcacggct ggtcgtctct tgtgggcggg 53820 caaagccagt atgtgggatc gtagcgccac cccaagtgtc catgccgtgg ttgtcgtaaa 53880 gctgggaaag accaaggctt gttcttgggc caaaacaagg gcgggctatc ccatctagct 53940 gattagccag aagttatcat attgtcggcg gggtgtgggc aaaggaagatt gattttctca 54000 cgtcagccgg gttatgcgat gaacaggct cttggggggc catggtagct gccggaatac 54060 ctgggtaatc cgagcgagaa tttgaagcgg tgaaagtcct taaggtcctt caagagcgct 54120 tccttcgact taagcagag ttcagagata gattgcaaac tgtcagtggg ccatcgtccg 54180 ggatcgtctg gctggagttt ttgatactcc gcattggagc aggtttagcg agaccatgct 54240 aaaagcttat tctgctgctc ctggtcctat cacgaattgc caagatgaac cgtcttaagg 54300 agatcgtgcc ctgccaagct gcctttgtgt your footcc acttacacta cccactatct 54360 cgtttcgtca tgactgtaca tcctcccgcc gtggcagacg agacctcccc actactgcca 54420 tcgcaagatg ggcctggaca taatggcatc gttcctgcag caaccaagcc caaagaactt 54480 caatcatcaa tgagtcaggt cgcgttactt tgttgtgctc gagcaattga cccgctggcc 54540 tttttcacta ttttccccgtt cgtcaaccag atgattgccg acacggcagg tatcgacgaa 54600 gcggacgtcg gcttctacag cggcatcatt gtcagccctc tacctcctct cttagtacca 54660 54720. ttcgctac aacttccacc ttcaggaatc gttgttttct gtgactcaga tgatgttaat gattccctgg gcacgagcag cagatcggat gggacgcag ccggtcctga tcctctccct 54780 ggccggttta tccgtctcat ctgcgctgtt cggattcagt cggactttag gccaaatggt 54840 tttcttcaga tgcttggcgg ggacgttcgg tgggaccgtt gtgtacgtga tatgctgaca cgttgggacg ggtgagacgc tgatcatgaa ttgtatagta cggtccgagt gatgatttcc 54960. 55020. ctccagacac ccaggcccgg gcattcagct acttctcctt ggcgaacacc attggaacgg tgattggccc tctgcttggt aagctaatgt ttatccctgc cggcagcacg 55080. cctgacacat gctgatgtgg gctccgtagg cggagcactt tgtcgtcctg gtggtgtgtt 55140 ccgtcactac ccgtatgcgt tgcccactgt cgcggccgga gcttttggag taactgtcac 55200 ggttgcctgt ctgatgttcg tcaacgaggt gagaggccat gggtaaggaa caatgacaca 55260 attcgcggaa gctaagatgg gtcctccaga cgcggaaacc cgcagatcat acacctcatg 55320 aaacagcctc gccaacatgg actagtgcca agatcctccg atcccaaggc gtgctaccag 55380 ttctctacat tcacggccat tcgatgatgt tggcattcgc gtacactgcg ggtaagctct 55440 ctttcgagcg tcccaaatat ctacaaaccc taaccagctc tcttattgaa gtttccccgg 55500 tattctactt cacatcacca cgtctaggcg gctacggttt ctctccattc tatatatcac 55560 tcttccttgg cggatcgggg atcgctcaaa ccatctggct agtgctagta tacccgccac 55620 tacaaaacg tctcggcacg ggtaatatct tacgaggctt atgtttcgtt tggattattt 55680 tcttggctgc tactgtagga gccagtgttc tccaccgtca tggtgagatg gtagcgttct 55740 ggattttggc gccgctagct ttggtactag gcagcagtgt tgcaatgcag ctgagtgaga 55800 cacatcccaa atttcctcac tactctatgg aggtggaggg ctgatagcta atggtattaa 55860 gcggctatgc aactcgcgct cgactccgtc tcaccgtcac ccgccgcctt aggcacgctc 55920 aatgccatgt ccttggccat catcagcttt cttcgcgctg tcgccccagc aatgttcact 55980 tccatgtacg caagtaccct taagcttagt agtcctggat tttacacatt ttggctcgtg 56040 ctgggtgggc tagttcttgt cctggcattt acactgcgct ggctgcctga gcaggtggag 56100 aaggctcctc gcaagctggg tcggtcgagc gcctgaggac agaagatatt atgatcgtgt 56160 tgtcgcatca gtgtcgacaa tgctgtccct tactatttat taacttgacg acaaatcact 56220 gtcccgggtt attagacatg ggtgttgaca tcttcgagtt tattgaacat agtaccaagg 56280 atacgcggta attagcggaa ataggtgcaa gccggaaccg tggctgaacc tcagagttca 56340 gaatgctgct agattggaac atactataca agcgaataac gtaccaatca cggtactatt 56400 tatgaacaag tatccaagaa tcaaatttgc gcaaggggga cagtaccact cgaccctaca 56460 tgatacagct caatgcgttt acgcagcgag atttaggatt tgaatcagac atcctacggg 56520 cgttctcagg cgctctacaa gggttttccg gtagtgaaaa gtactatgga atacccttcc 56580 cccttttga agcaaggtta ttatggcact cgatgcatgg gagataccag agaagggctt 56640 ctgatgcagg tgacgtgttc cattcatagc cctcgtccta atattggt gatatcgagc 56700 ttctggcatg ttatgacgat gcttactctg ttcacttgc taggtgggga attccagacc 56760 tcgaatacgg aggacccaag agttatcgta cttgagaag gatctgtaca tcccgaatgg 56820 gtcacccat agtcagactt ccggtatata cacactgctt gttatgtttc 56880 aattcaacga ccaaagact cgcctgatga aaaaatcaga ttggacgtct gaacatgtaa 56940 gatatagaat gagctccagg aaccagagct tcgattggga ttcatcact ataagtttaa 57000 gcccctggct cgagcatgtg actatgtaa taggttgagt agatctact gtcgatctgt 57060 caacggcgcc gacgacgact ccgttgcatc gccttgtagt taaccttaa cgttaataa 57120 atcttgaata agccgaaca aaacttatat tcattatatg taaatagga gatgtgata 57180 gtcgttactc actctgcaaa tctaacacc cgatgcaaaa gccaattaat gccgcggtat 57240 ggacaccggc acactgata tatatacaca tgttacgcag caacgatct agggcaacgg 57300 caccggataa ttgaactga aaagctcatc cggatcccac ttagactca actgcgagag 57360 cttaggcagg tgttgtggcg agtaccaaac atcagcgcct tcatcgccat gagcgtaatt caagtagtc tccatctcgc taaagccact agtacgctgg aactgtgagc ggatggaccg catgagga ttgacggcgt tatcgagggt agaattatca ctggggtagg agttttcgaa taacctatac ccagtgtgtt agtcacgga tgaatcgtgt tggatggcag aagcatacag atgagaattg atgtttcggt agccatagcc gtaggaggtg ctgttggctg gaacggctgc cgccgctgct gaactgtagc ggtcgacaac gaattccc gaatcgg ggtgagcctg ggagaaggcc tgcagctcat tgaagaaggt tgtgtaggtg tcgatgtttg tctcggccag 57780. tcccagtccc caggtgttga gataggaacc ggtggtgcag gcatcaccgt ccgctccaaa ggccatcgag cttgtcacgt tgggccaggc taccatcatt agattggttt ctgttgcatt 57900. gagcgctgta aaattggtta ggtaagatac tccctcttct tttggcccgt agtagcgc 57960 attack agctccatct gtcaccccgt ttgggttagt cttgaaaatc atgaaagtga ggctacgtgt gaacgaacct cttgagtggt ttgattgaat cccgagtatg ccgtcaatga 58080 cagctccggg ggaagcgtct cgtcaaaaga ctggaagatc tcccatacgg acctattctc 58140 gctagcgggg aataggtaat tcgcgctcat ggccaggccg ttatttgtgg cattgtagat 58200 cttgtaagtg gctgaggtga tgatgccaaa gttggcaccg gcgccgcgga tggcccagaa 58260 gagatcggca ttttcagagg tagatacagt gaggacatca ccgctgggcg ttaccagctc 58320 gacggacagt agggagtcga gaatcagccc atgcaggcct tgtagatttc ctatcccgcc 58380 gccgatagtc gccccagcga gtccgacaca cggtgcggta ccggtagctg ggtaaatgct 58440 gttagcatag ggctgtagac attttggatg ctggggggga gagcaagcag catacgcagc 58500 tctttacccg cctcatagag aggcggaatg atgtcctcaa agcgtactgc tccaccaacg 58560 gtcatggtat tgttggaagc gtcaatcgaa acagtattga agttgctaat atcaatttcg 58620 atgccattgg tacaattaac ataggtgatg gtcgcgccgt ggccgcctgc tgtggcgaga 58680 aaagggatct tgttggatgc agctagggtc acctagtccg caattttgcg gtgagctgtg 58740 gtacttgttg tctgataatt gaatggaagg cgtcgtctta caatgttttg tatatctttg 58800 accgtagctg gcttgattgc cccgatgtag gtgggggcgt cgtatgtggt ccatctctgt 58860 gttacgtcct ctgtgtagtt tgtgtcggaa ggtagaagaa tctgcgcgcc cggagacagg 58920 ctggaactga agatggatgt gatgttgtgc gtggcagcag acgagagtgt ggttagcccc 58980 agagcaaa gaagaattgt ccgaaacatg attggccgac gagactgaa cgagaagaca 59040 aacggcgaga cattttggaa taggtcccccc ttatcagtag cgtactatat atttttgagt 59100 cctggatttc caatgatggg gattatgcta ctccgaggga agcgcaataa ctccgtgctt 59160 atgcgtattg tcagaccggc ttgagagtga ctggaaggat gcgggtccct gttggctagc 59220 aaaagtccga agggaaactc catcgcatgg ctgcaagtgt gattatctca gcctaagagc 59280 actctgcttt cacgcgagag gtacgttaag ccagtgtttc agaggcgagg agccacgga 59340 gaactgcagg gtcgtccgat gcactgcgga gtaattcgcc ggacttgccc taaaaggaa 59400 gtagaaccag gacaatctcc atgtggttgg tacggcagag tatcatatcc gccaagagag 59460 tcaacgagga tcaactgatg gaatataatg aaaataggct tcatggccct atccaaaaat 59520 cagcccagtc cagaacggtg atttcccctc cattcgacgc tacatacccc tcttgaacat 59580 taacgtagat tgtacattct tttctcttca gacctgtgag cgatttaccc acttacaatg 59640 gccggagagt attcaacggc cccgatggcc atcataggca tggcctgccg cttctctggc 59700 ggcgccacga gccccgagaa gttgtgggac atgatcgtac aacggcgatc cggctggtcg 59760 gagataccaa cctcacggtt taacgcaaat ggactttacc atccaaacgg cgagcgtgtg 59820 gggacggtaa gacatgctta ttctgtccct tcaagtgatc aagttgaccc gtgcctttct 59880 ctctagaccc acgtgaaagg tggtcacttt ctcgaagatg atattgcctg cttcgatgcg 59940 gccttcttcg gcatggcctc agaaaccgct agtgtaggat tgttccgtcg aattctccca 60000 gggggcctac caccaaaatg aactgacatg aaggctgcta ggctatggac ccgcaatatc 60060 ggatggaact cgaggttgtc tacgaggctc tggagagcgg tatgcagtca cctttcacgc 60120 ctatcaccgc ccgaggactg acacatttgc ttgcagcggg gattccaatg gagtccatta 60180 aaagaacgaa tacttcagta tatgggggtg tgatgtttcg cgactaccac gacactcata 60240 gccgtgactt ggaccgcta cctcgatact ttatgacagg caatgctgcc acgatggcgt 60300 cgaaccgcat ttcccacttt tatgaccttc gcgggccgag catgaccgtt gatacaggtt 60360 gctccacaag tctgacggct ctgcacctgg cttgccagaa tttgcgcagt ggagaatcca 60420 acatgtctat cgtgacggga gcaagtctca tgataaaccc cgacgttttc ctcagcatgt 60480 ccaatatcgg gtgagtaact tctagctgtc acacgttgaa caaatggcac tgataaatgt 60540 gaccaagctt tttgtcgccc gacggtatat cgtatgcctt tgacagccgt gccaacgggt 60600 acgggagagg tgaaggagtg ggtgccttac tcgttaagcg attagacgat gctctgcgcg 60660 atggcgactc catccgcgcc atcatccgcg aaaccggggt gaaccagaac ggtaagaccc 60720 ccagcatcac agccccacaa caagctgccc aggaagcttt gatccgccag tgttatgaac 60780 gggtcaactt ggatcccgca cagaccacat acgttgaggc tcacggacc ggtactccag 60840 ccggcgatcc gctggaggtc ggcgcgctgg cagccgcatt gggaggcagt cgatcggcgg 60900 agcacccgtt gtatctcggc tctatcagg ccaatattgg accacactgag gctgccagtg 60960 gcgtggccag catcatcaag gtggccctag cgctggagaa gggacaaatt ccgccgaata 61020 cgcaactgaa tacaccgaac agtgagctgc ggttaatga tagaacatg gaggtaggct 61080 acaatacctt atttggtaga aactctcat gcgaacggtt atgtttgaac aggttcctgt 61140 atcaacgcag agatggcctg tgggaagg tccccgacgt gcctccgtca acaatttgg 61200 ttttggcggt agcaatgctc atgccatttt ggaatcgccc ccggtaggtt ccaccaacgg 61260 cacccatgta atggaacgc atacaacaa cggaacggct gggatgaacg gcacaactgg 61320 agaaaatggg gcaatcgaa ctaacggggca gctaagagc aatggtccgg tggtaaatgg 61380 caacaaacc aacattacca aaagggaaac cccatgggtg ttccgcttga gtgcaagga 61440 tgcccagact tgccagcaga tggccgccga cctaagtacc tatatagagt cccaccacc 61500 agtcgacgag gaagccttcc tgggccgcct tgcatacacc ctgggatcgc gccgttctgt 61560 cttctcatgg accgcagctg tgtcggctag gtctctggca gagttaacac gggcactgga 61620 cgatgatgaa cgtttggtgc cgtcacgcgc agccccttct ttacgattgg gatgggtatt 61680 cactggccaa ggggctcagt ggtatgctat gggtcgtgaa ctaattgcca cttacccggt 61740 atttcggtca accattctgg agtgcgaccg ctacatgacg gagatggggt cgacgtggac 61800 attaatgggt gagtattgag cgctctatct tgaagcaacc tttgctaatt atacataaag 61860 aggaactgca tagagaggaa tccacttcac aggtcaacaa catcgtctac agtctgggtc 61920 tggccaccgc catccagata gccctggtgg agctgctatg gtcctggggc atccatccca 61980 cggctgtgac aggtcattcg agcggtgaga ttgcagcagc gtatgcatcg aaagccctgg 62040 acatgaagtc agccattggc attgcttacc ttcgtggagt gttagctgag aagtttgatg 62100 acaagattct cggcaaaggg ggcatgatgg ccgttgggct tggacgtaaa ccggttgaac 62160 attaccttag tcgcgtcact gcggggtact gcgtcgtggc ctgcgtaaac agccagtata 62220 gcgtcacaat atcaggtgat atcccagcaa tcgaccagct tgaacagttg ctgcaggaag 62280 accaagtctt cgctcggcgt ctccgcgtca atggggcgtt tcactgcgag cagatgcgcc 62340 ctatggctga cctattcgat tggagtctac gctacctgct cacccctcat cccgacttcg 62400 gatcagtcct attctcctcc cccaagacag gctcgcgcat ccaagacggc actatactag 62460 cgacctcgtc gcactgggtg ggaaatatgt tgcaggcggt tgagttcgag agctccttcc 62520 gtcatatgtg ttttggtgat ccaagcccaa aaggcgccaa aggcacccaa gatgtcgacc 62580 tagtactcga gatcgggcca catggagcgt tagggggtcc gattcaacag ctcatgaccc 62640 tccccgaatt tgaaggtagt ggcatttctt acctgcccac tctggtacga aagcaggatg 62700 ccgtcttcgc catgcagcgc ctggcgattg acctaactca cagaggctac cccgtcgatc 62760 tcaatgccgt caatttcccc cacggcacgc tctctctttc catcttgcac gaccttccat 62820 catacccatg gaatcatagc acgagatact ggcttgaacc ccggcgaaac cgagccgaca 62880 gacagcggca agcgccgccg tcggatttgg taggttactc tcaaccaagc atcacgccac 62940 ttgcacgcac ttggcgccac attattcgct tatctgatct gccctggctg ggagatcaca 63000 gggtgcagtc gagcattgtc ttccccggtg ctggcctggt gagtatggcc atagagggca 63060 tgcggcaagt ggcggcgggc cgccagcaaa ccgtgtccgc gtacgagctg cgcgacgtag 63120 atattgccaa agccctcacg gtgcctgagg cagatgaggg agtcgaagtg caattaaata 63180 tccgcccgtg tgatgagcaa atgctgggga cgaaggactg gctggctttc cagatcttct 63240 ccgtctctgg cgacagccgc tggacggagc actgcagtgg acgaatttca gttatcacaa 63300 cctctgacag cactcctctc ccatctgcca taccatccca gtcggaggac ctgtataacc 63360 gccgcattga tccccgctat atgtgggcag ccatgcgatc cgtgggcata taccatgggc 63420 ctctgtttca aaacatccac caagttctgg cgaagccgtc cgcatctcgc accatttttg 63480 ctatcgcaga tacggctgcc gtgatgccga agaagtacca gaccccccat gtgttgcatc 63540 ccaccacgct ggactcggtc ttccaggctg cctatacact tttgcccgag tctggcgcgc 63600 gtctgccttc cgccatggtt cctcgccata tccggtctgt ccgagtttca gcgcagatct 63660 ccaatagtcc agcgcatgac ctggaggcat atgccacgct gaatcgggat tatgacgcgc 63720 agtcattcga gacatccctc actgtggtgg acgcaaagga cgggaacagc ccggtgctcg 63780 aagtggatgg cttgacctgt cagtcacttg gacgagccct ggatcgcgag gcagatccgc 63840 acgagaatga aatctgcagc cggtgggaat gggctccaga tattggcaca cttgatgcgg 63900 ccgcctgcaa ggaccggatt cgctgtgcgc cagaggcagc ggagatagag acgatgcgcg 63960 acctccgccg agccactatt ctatacattc ttgacattgt ctccagcctg acggtggctg 64020 atgttcagca gttgcgggga catcttaaga aattctacgt gtggatggtg gagcagctga 64080 aaaaagccag tcgcaatcag ttcgcgcccg acagcgccca gtggcgcgat atcagcgcgg 64140 cggacaaggc agccctgtac gagaaggtcg gtcggaccag tgtgaatggt gagatgctct 64200 gccgattggg tccacttgga gcatcttttc tacggcaaga aatggctcct ttagaggtaa 64260 tgttggaaaa ccggctttta ttccgctact acctcgaggc gctaaaatgg gatcggtcga 64320 cacgccaggt aagtgagctc gtcaggctct gcacgcacaa gaatccccga gccaagattc 64380 tggaaatcgg agccggcaca ggaggcggta cgcaggtgat cttggaggcc ctgggcaaag 64440 agaatggttc ctcgacgggc gctcgatttg gccgatatga cttcacggat atctctgcgg 64500 gcttcttcga ggctgccaaa gagcgcttcc aagactgggc ggacttgatg aacttccaga 64560 agctagatat tgaacacgat cccgtggctc aagggtttga agaaggctcc tatgatgtcg 64620 taattgcctg ccaggtgctt catgccacca agtcgatgga tcgtactctc acacacgttc 64680 gcaagcttct gaagccaggg ggcaaattga ttctaatgga gaccacccgc gacgaactag 64740 atgtcttctt cgcattcggc cttttgccag gatggtggct gagtattacc tccccttcct 64800 ggcacttctc attatatttg gtatatggca ctaattgtgt tctttaacag gcgaggaaga 64860 ggaacgtcga actaccccat cactcaccct gccattctgg aaccaggttc tatcccgtaa 64920 cggcttcgcc gggttggacc ttgaggtcca cgactgcgac agtgaggaat tctacgcttt 64980 cagcacaatt ctctccaccg ctcaagcccc tgctttatct ataacatcac ccgtaactat 65040 cgtcactggc acatcgccac cgccgacatc ttggatgagc gagcttcaaa cagccgtggc 65100 agctcatatc ggatgtcaac cagtaattgc gacgttggag acagttaccc cccagggtaa 65160 tatctgtatc tttctcggcg aggcggacga gcccctgctg gaccatgtca gtaacccagt 65220 cgaatttgac cgtatcatac acctggccac aagatgcaag ggcctcctgt ggatcacccg 65280 cggcgggtct ttggacgtcg acaaaccggc catgagtctg tcccaagggt tactgcgcac 65340 cctgaagtca gaatatcaag gcaaatcctt tgtctccttg gacgtggatc cacgacggtc 65400 tccatggact gcagaagtag tccaagccat atcccagatt tttcccgctt cgttcagtga 65460 gactactgac cctgccacgt gtgagttcga gtatgcagag cgcgatgggg tcctacacat 65520 tccccgtacc gtgaaggata tccccatgaa cagaaacatt ttccctgaga gcgacacgac 65580 tgagaagacc atacattgcc ggttccgaga tgcggcccgt ccccttcgca tgaagatcgg 65640 gaccccaggg ctcatcgaca ccttggtctt ccacgacgat ttagacgcaa aatccgaccc 65700 gttgcctgca gactggattg aattcgaccc cacggctttt ggtctaaact tccgcgatgt 65760 catggtagca atgggccagt tggaggccaa tgccatcatg ggcttcgagt gcgcgggaac 65820 catcgtccgg ctgggagcca ctgcggcagc gaagggcttt gcagttggcg atcgtgtatg 65880 tacgctactc aggggccact gggcgacgcg gcctcgagca ccctggacca gcgtgatgag 65940 gatccctcaa catctaagcg accaggaggc cgcctctttt cctactgtct ttgctaccgc 66000 ctacatcgcc ctgcacgaaa ctgcccgcct ccaacgcggc gagtcgatcc tcattcacgc 66060 tgctaccgga ggagtcggcc aggctgcgat ccagcttgcc caactaatcg gggcagagat 66120 ctacgccacc gcgagtactc cggcaaagcg gcaattgctc catgagacgt atggtattcc 66180 agagaacaat atcttctcca gtcgagatcc atcattcgcc accgatgtac atttgcggac 66240 cgacggccga ggcgttgacg tcgtattgaa ctcccttgca ggccgtctgt tacaggagag 66300 cttcaactgt ttggcggaat tcggtcgcat ggtagagatc ggcaagcggg atctcgagca 66360 acacagcggc ttagacatgt acccgttcac ccggaacgtc tctttctcgt cggtggactt 66420 gctgacatgg cagagccgga gaggcgcaga tatctcttgt gtcctgcaaa gcctctcgaa 66480 gctactgggc gagaagaaga tcatgccggt ctatcccctt acactatatc ctatcacaca 66540 gatcgaaaaa gccttccgaa ctatgcagac tggccaacac atgggcaaga tcattatctc 66600 ggtcggcgaa caggatactg tgccggtaag tcgcactctt cccagttcca acctcacctc 66660 gactgcaggt gaattcgaca agggctaatg tgacctgggc aggtggtgga gcgtccccca 66720 cctttctctt tgcgatccga cgcatcgtat gtcattgtcg gcggccttgg cggtattgga 66780 cgcgtcctgt gtgaatggat gatggcacgc ggggcacggc atctgatcat catatcgcgg 66840 aatgctcgcc ctgggccttt tgtgactgag cttgagcaac aagggtgtga agttcgcacc 66900 ctcgcctgtg acattgcggc agaggatcag ctggcggcag cactggcaca gtgtgctgac 66960 atgcctccta ttaaaggcgt catccagggc gctatggtgc tcaaagtatg ttgcctccct 67020 tcatttcata tcgatctcta gtggatgggt attgcagcat atactaacat ttacatagga 67080 taccgttctc gagcaaatga cagtcggcga cttcgaagcg gcagtacgcc ccaaagcgca 67140 gggaagctgg aacctgcacc agcagctggg cgacgttgac ttcttcataa tgctttcttc 67200 cttgatggga gttatgggag ccgccagcca agcaaactac gccgccgggg gtgccttcca 67260 ggatgccctg gccacgtacc gccgcaaccg gggtctgccc gcagtgagtc tggatctggg 67320 aattgtccga tcagtagggt tcgtagccca gaccgatggc gtgcaagagc gtctcgtcca 67380 gatgggggtt acctctctgt ctgaagaaac agttctccgg atcctcgaac aagccatcac 67440 gcatcccaca ggtccacccc agatcatcac cggtattaac accgccccag gtaaacactg 67500 ggacgaagcc tcctggatcc aagacccgcg ctttgccgct ctgagatacc gcgacagtac 67560 gcaggctgga tcttcccgtg ctactactgg gacggcgaaa cagggaaaga taagggatca 67620 gcttgcagag attgcttccc cagtggatgc tgctgcactt atctgccagg agttgatgca 67680 gaaattagcc agcatgtttg gtctggtggt cgaggagatg tctgctactc aagatctgtc 67740 ttcctatgga gtggactcgc tggtagcggt ggaactgcgc aattggcttg tcgcgcaggt 67800 gggtgctgag gtttccatct ttgatttgat gcagagtccg tctttggagg acttatccct 67860 gcgggtggca acgaagcgga cgtag 67885 <210> 6 <211> 524 <212> PRT <213> Aspergillus niger <400> 6 Met Ser Ser Thr Thr Ser Ser Ser Arg Ser Asp Leu Glu Lys Val Pro 1 5 10 15 Val Pro Gln Val Ile Pro Arg Asp Ser Asp Ser Asp Lys Gly Ser Leu 20 25 30 Ser Pro Glu Pro Ser Thr Leu Glu Ala Gln Ser Ser Glu Lys Pro Pro 35 40 45 His His Ile Phe Thr Arg Ser Arg Lys Leu Gln Met Val Cys Ile Val 50 55 60 Ser Leu Ala Ala Ile Phe Ser Pro Leu Ser Ser Asn Ile Tyr Phe Pro 65 70 75 80 Ala Leu Asp Asp Val Ser Lys Ser Leu Asn Ile Ser Met Ser Leu Ala 85 90 95 Thr Leu Thr Ile Thr Val Tyr Met Ile Val Gln Gly Leu Ala Pro Ser 100 105 110 Phe Trp Gly Ser Met Ser Asp Ala Thr Gly Arg Arg Pro Val Phe Ile 115 120 125 Gly Thr Phe Ile Val Tyr Leu Val Ala Asn Ile Ala Leu Ala Glu Ser 130 135 140 Lys Asn Tyr Gly Glu Leu Met Ala Phe Arg Ala Leu Gln Ala Ala Gly 145 150 155 160 Ser Ala Ala Thr Ile Ser Ile Gly Ala Gly Val Ile Gly Asp Ile Thr 165 170 175 Asn Ser Glu Glu Arg Gly Ser Leu Val Gly Ile Phe Gly Gly Val Arg 180 185 190 Met Leu Gly Gln Gly Ile Gly Pro Val Phe Gly Gly Ile Phe Thr Gln 195 200 205 Tyr Leu Gly Tyr Arg Ser Ile Phe Trp Phe Leu Thr Ile Ala Gly Gly 210 215 220 Val Ser Leu Leu Ser Ile Leu Val Leu Leu Pro Glu Thr Leu Arg Pro 225 230 235 240 Ile Ala Gly Asn Gly Thr Val Lys Leu Asn Gly Ile His Lys Pro Phe 245 250 255 Ile Tyr Thr Ile Thr Gly Gln Thr Gly Val Val Glu Gly Ala Gln Pro 260 265 270 Glu Ala Lys Lys Thr Lys Thr Ser Trp Lys Ser Val Phe Ala Pro Leu 275 280 285 Thr Phe Leu Val Glu Lys Asp Val Phe Ile Thr Leu Phe Phe Gly Ser 290 295 300 Ile Val Tyr Thr Val Trp Ser Met Val Thr Ser Ser Thr Thr Asp Leu 305 310 315 320 Phe Ser Glu Val Tyr Gly Leu Ser Ser Leu Asp Ile Gly Leu Thr Phe 325 330 335 Leu Gly Asn Gly Phe Gly Cys Met Ser Gly Ser Tyr Leu Val Gly Tyr 340 345 350 Leu Met Asp Tyr Asn His Arg Leu Thr Glu Arg Glu Tyr Cys Glu Lys 355 360 365 His Gly Tyr Pro Ala Gly Thr Arg Val Asn Leu Lys Ser His Pro Asp 370 375 380 Phe Pro Ile Glu Val Ala Arg Met Arg Asn Thr Trp Trp Val Ile Ala 385 390 395 400 Ile Phe Ile Val Thr Val Ala Leu Tyr Gly Val Ser Leu Arg Thr His 405 410 415 Leu Ala Val Pro Ile Leu Gln Tyr Phe Ile Phe Cys Ser Thr 420 425 430 Gly Leu Phe Thr Ile Asn Ser Ala Leu Val Ile Asp Leu Tyr Pro Gly 435 440 445 Path Ser Path Ser Path Thr Path Val Asn Asn Leu Met Arg Cys Leu Leu 450 455 460 Gly Ala Gly Gly Val Ala Ile Val Gln Pro Ile Leu Asp Ala Leu Lys 465 470 475 480 Pro Asp Tyr Thr Phe Leu Leu Leu Ala Gly Ile Thr Leu Val Met Thr 485 490 495 Pro Leu Leu Tyr Val Glu Asp Arg Trp Gly Pro Gly Trp Arg His Path 500 505 510 Arg Glu Arg Arg Leu Lys Ala Lys Ala Asn Gly Asn 515 520 <210> 7 <211> 24 <212> DNA <213> Artificial Sequence <400> 7 caccaggtcc tcagatactg bag 24 <210> 8 <211> 24 <212> DNA <213> Artificial Sequence <400> 8 aaactctaca gtatctgagg acct 24 <210> 9 <211> twenty four <212> DNA <213> Artificial Sequence <400> 9 caccgagacg caagccattg ttat 24 <210> 10 <211> twenty four <212> DNA <213> Artificial Sequence <400> 10 aaacataaca atggcttgcg tctc 24 <210> 11 <211> twenty four <212> DNA <213> Artificial Sequence <400> 11 caccacctac attacagctg ccgt 24 <210> 12 <211> twenty four <212> DNA <213> Artificial Sequence <400> 12 aaacacggca gctgtaatgt aggt 24 <210> 13 <211> twenty four <212> DNA <213> Artificial Sequence <400> 13 caccaagtca gcgaaggata cttc 24 <210> 14 <211> twenty four <212> DNA <213> Artificial Sequence <400> 14 aaacgaagta tccttcgctg actt 24 <210> 15 <211> twenty four <212> DNA <213> Artificial Sequence <400> 15 caccctcgat aggtggttcg agac 24 <210> 16 <211> twenty four <212> DNA <213> Artificial Sequence <400> 16 aaacgtctcg aaccacctat cgag 24 <210> 17 <211> twenty four <212> DNA <213> Artificial Sequence <400> 17 caccttcttg cgcatgtcgt gtgg 24 <210> 18 <211> twenty four <212> DNA <213> Artificial Sequence <400> 18 aaacccacac gacatgcgca agaa 24 <210> 19 <211> twenty four <212> DNA <213> Artificial Sequence <400> 19 caccagtcac ggctatgagt gtcg 24 <210> 20 <211> twenty four <212> DNA <213> Artificial Sequence <400> 20 aaaccgacac tcatagccgt gact 24 <210> twenty one <211> twenty four <212> DNA <213> Artificial Sequence <400> twenty one cacctatctc agagcggcaa agcg 24 <210> twenty two <211> twenty four <212> DNA <213> Artificial Sequence <400> twenty two aaaccgcttt gccgctctga gata 24 <210> twenty three <211> twenty one <212> DNA <213> Artificial Sequence <400> twenty three ggttggagat tccagactca g 21 <210> twenty four <211> twenty four <212> DNA <213> Artificial Sequence <400> twenty four aaaaaagcac cgactcggtg ccac 24 <210> 25 <211> 57 <212> DNA <213> Artificial Sequence <400> 25 aaatagtaga aatggcattg agatccacat gctctatact caggaaacag ctatgac 57 <210> 26 <211> 58 <212> DNA <213> Artificial Sequence <400> 26 ccatgcgcgt gttataaacg aacttgatct gcagcgagtc tgtaaaacga cggccagt 58 <210> 27 <211> 57 <212> DNA <213> Artificial Sequence <400> 27 cattgatcgc tgcatatgga tctgttaata cgacgatctc caggaaacag ctatgac 57 <210> 28 <211> 58 <212> DNA <213> Artificial Sequence <400> 28 agaccttcgc aatgatcggt gactttgtca accggaacac tgtaaaacga cggccagt 58 <210> 29 <211> 57 <212> DNA <213> Artificial Sequence <400> 29 tccacagtat ggagcgctgg aaggcaagca gttgctgcgg caggaaacag ctatgac 57 <210> 30 <211> 58 <212> DNA <213> Artificial Sequence <400> 30 ttggcgaatg ctcacacgct cgcttcgtat ccaacaacag tgtaaaacga cggccagt 58 <210> 31 <211> 57 <212> DNA <213> Artificial Sequence <400> 31 cgaatacttc agtatatggg ggtgtgatgt ttcgcgacta caggaaacag ctatgac 57 <210> 32 <211> 58 <212> DNA <213> Artificial Sequence <400> 32 cagtagtagc acgggaagat ccagcctgcg tactgtcgcg tgtaaaacga cggccagt 58 <210> 33 <211> twenty three <212> DNA <213> Artificial Sequence <400> 33 gatgacacca ccatagtcac aag 23 <210> 34 <211> twenty three <212> DNA <213> Artificial Sequence <400> 34 cgagtccaag tgaatgttga atc ​​23 <210> 35 <211> twenty four <212> DNA <213> Artificial Sequence <400> 35 atgatgaccg tcaagacgcc gttg 24 <210> 36 <211> twenty four <212> DNA <213> Artificial Sequence <400> 36 atggggttat cgcacagact gttc 24 <210> 37 <211> twenty four <212> DNA <213> Artificial Sequence <400> 37 gtcacaacga caagggctac taac 24 <210> 38 <211> twenty four <212> DNA <213> Artificial Sequence <400> 38 aacctgactg aagcaatctc cgag 24 <210> 39 <211> twenty two <212> DNA <213> Artificial Sequence <400> 39 agtgctatgg acccgcaata tc 22 <210> 40 <211> twenty two <212> DNA <213> Artificial Sequence <400> 40 ggcttggact acttctgcag tc 22

Claims

1. Use of an endogenous active polypeptide in the production of citric acid by Aspergillus niger fermentation, characterized in that: The amino acid sequence of the polypeptide is shown in SEQ ID NO: 1, and the activity of the endogenous active polypeptide in Aspergillus niger is weakened or completely removed.

2. Use of a gene cluster in the production of citric acid by Aspergillus niger fermentation, characterized in that: The nucleotide sequence of the gene cluster is shown in SEQ ID NO: 5; the entire gene cluster is inactivated in the Aspergillus niger.

3. A citric acid producing strain, which belongs to Aspergillus niger, characterized in that: The strains were modified: 1) weakening or completely removing the activity of an endogenous active polypeptide, wherein the amino acid sequence of the endogenous active polypeptide is as shown in SEQ ID NO: 1; or, 2) inactivating a gene cluster, the nucleotide sequence of the gene cluster is shown in SEQ ID NO:

5.

4. The production strain according to claim 3, characterized in that The activity reduction or complete removal of the endogenous polypeptide can be achieved by one or a combination of the following methods: partial or complete knockout of the polypeptide encoding gene; mutation of the encoding gene; changing the promoter, translation regulatory region or coding region codon of the encoding gene to weaken its transcription or translation; changing the coding gene sequence to weaken the stability of its mRNA or destabilize the structure of the encoded protein; or any other method of inactivating the gene by modifying the gene coding region and its adjacent upstream and downstream regions.

5. The production strain according to claim 4, characterized in that The activity of the endogenous active polypeptide in the strain is weakened relative to that of the unmodified endogenous active polypeptide, which means that the transcription and expression of the coding gene of the endogenous active polypeptide completely disappear.

6. The production strain according to any one of claims 3 to 5, characterized in that The activity of the citrate transporter in the production strain is enhanced, and the citrate transporter is a polypeptide with an amino acid sequence as shown in SEQ ID NO:

6.

7. A method for constructing a citric acid producing strain, wherein the strain is Aspergillus niger, characterized in that: The method comprises genetically modifying: 1) causing the activity of an endogenous active polypeptide in the strain to be weakened or completely removed relative to the activity of an unmodified protein, wherein the amino acid sequence of the endogenous active polypeptide is as shown in SEQ ID NO: 1; or, 2) inactivating a gene cluster, the nucleotide sequence of the gene cluster is shown in SEQ ID NO:

5.

8. The construction method according to claim 7, characterized in that: The activity reduction or complete removal of the endogenous polypeptide can be achieved by one or a combination of the following methods: partial or complete knockout of the polypeptide encoding gene; mutation of the encoding gene; changing the promoter, translation regulatory region or coding region codon of the encoding gene to weaken its transcription or translation; changing the coding gene sequence to weaken the stability of its mRNA or destabilize the structure of the encoded protein; or any other method of inactivating the gene by modifying the gene coding region and its adjacent upstream and downstream regions.

9. The construction method according to claim 8, characterized in that: In the construction method, the activity of the endogenous active polypeptide in the strain is weakened relative to that of the unmodified protein, which means that the transcription and expression of the coding gene of the endogenous active polypeptide completely disappear.

10. The construction method according to any one of claims 7 to 9, characterized in that: The activity of the citrate transporter in the production strain is enhanced, and the citrate transporter is a polypeptide with an amino acid sequence as shown in SEQ ID NO:

6.

11. A method for producing citric acid, characterized in that: The method comprises: (1) culturing the citric acid producing strain according to any one of claims 3 to 6, or the citric acid producing strain obtained by the construction method according to any one of claims 7 to 10, so that the strain produces citric acid; and (2) Optionally separating citric acid from step (1).

12. Use of the citric acid producing strain according to any one of claims 3 to 6 or the citric acid producing strain obtained by the construction method according to any one of claims 7 to 10 in producing citric acid.

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