CN113088533B - Yeast engineering bacterium for efficiently expressing barnacle viscose protein and preparation method thereof - Google Patents

Abstract

The invention discloses a vector for expressing viscose protein, yeast engineering bacteria and a method, belonging to the technical field of protein preparation. The invention is based on a pichia pastoris expression system, optimizes the parameters of initial pH value, inoculation quantity, methanol addition quantity, induction time, induction temperature and the like of a bacterial fluid culture medium by technical means of increasing copy number, fusing lipase, increasing auxiliary factors and the like, improves the expression quantity of the mucin to gram level, and meets the requirement of large-scale production and application. Furthermore, the invention also discloses an expression vector and a yeast engineering bacterium for expressing the viscose protein.

Description

Yeast engineering bacterium for efficiently expressing barnacle viscose protein and preparation method thereof

Technical Field

The invention discloses a vector for expressing viscose protein, yeast engineering bacteria and a method, belonging to the technical field of protein preparation.

Background

The gels commercially used at present have strong adhesive properties in dry environments, but have greatly reduced adhesive strength and mechanical properties in wet environments. Therefore, the development of hydrogels that are compatible with aqueous environments, have both excellent adhesive strength and excellent mechanical properties, is a challenging desideratum. Although there are biomimetic hydrogels developed based on DOPA in mussel byssus protein, the balance of the redox system in which DOPA adhesion occurs is more difficult to control in the extracorporeal environment, thereby affecting the adhesion effect of the hydrogel. The adhesion mechanism of the barnacle gum is different from that of mussel byssus protein, and the barnacle gum is directly adhered without any modification. Therefore, the barnacle viscose protein is used as a base material, the bionic structure hydrogel with better performance can be developed, the problems are solved, and the method has important research value and huge application prospect.

Barnacle gum is composed of a large amount of protein and a smaller amount of carbohydrates and lipids. According to the difference of protein molecular weight, the barnacle viscose protein is divided into six components, namely cp16k, cp19k, cp20k, cp52k, cp68k and cp100k. Wherein cp19k and cp20k are key proteins in interfacial adhesion, and recombinant Mrcp19k from the red giant barnacle can adhere to the surfaces of various materials; the recombinant Mrcp20k adheres to the material surface of calcium carbonate and metal oxide. Both proteins show good surface adhesion.

At present, there are two main methods for obtaining barnacle viscose protein components, namely a chemical extraction method and an in vitro recombinant expression method by using an escherichia coli prokaryotic expression system. The component of the barnacle albumin separated by the chemical method has low purity, the required sample amount is large, and the preparation process is complex. In the existing research, a escherichia coli expression system is mostly adopted to obtain the recombinant barnacle viscose protein, the recombinant barnacle viscose protein is influenced by the expression system, the expression efficiency and the expression level are limited, and inclusion bodies often appear. Therefore, on one hand, the functional difference between the recombinant protein and the natural barnacle viscose protein is caused, and on the other hand, the recombinant protein is not suitable for large-scale production and engineering application.

The pichia pastoris expression system is one of the most widely applied eukaryotic expression systems at present, however, no technical data about the expression of the barnacle mucin cp19k and cp20k in pichia pastoris exists at present. Although data are related to the expression of other barnacle mucilaginous proteins (e.g., cp52 k) (inspired. Heterologous expression and functional study of the 52kDa glue protein of red giant barnacle [ D ]. National defense science and technology university, 2016), the expression efficiency is low and cannot meet the requirement of large-scale production. The technical problem to be solved by the invention is to efficiently express barnacle viscose proteins cp19k and cp20k in pichia pastoris and provide a method capable of efficiently expressing the viscose proteins.

Disclosure of Invention

In order to solve the technical problems, the invention is based on a pichia pastoris expression system, optimizes parameters such as initial pH value, inoculation amount, methanol addition amount, induction time, induction temperature and the like of a bacteria liquid culture medium by technical means such as copy number increase, lipase fusion, auxiliary factor increase and the like, improves the expression amount of the mucin to gram level, and meets the requirements of large-scale production and application.

The invention provides an expression vector, which is characterized by comprising any one of the following components:

i) The vector pAO alpha N-4Mrcp19k has a sequence shown in SEQ ID NO. 1;

ii) pAO alpha N-4Mrcp20k vector with the sequence shown in SEQ ID NO. 2;

iii) The vector pAO alpha N-4proROL-Mrcp19k has a sequence shown in SEQ ID NO. 3;

iv) pAO alpha N-4proROL-Mrcp20k vector, the sequence is shown in SEQ ID NO. 4.

The invention also provides a vector combination, which is characterized by being formed by combining the vector with pPICZ-Ssa4-VHb and pPIC3.5k-Sso2-Bmh vectors; wherein the sequence of pPICZ-Ssa4-VHb is shown in SEQ ID NO. 5; the sequence of the pPIC3.5k-Sso2-Bmh vector is shown in SEQ ID NO. 6.

The invention also provides a yeast engineering bacterium which is characterized by containing the expression vector or the vector combination.

In some embodiments, the engineered yeast strain is pichia pastoris GS115.

In some embodiments, the engineered yeast strain is deposited in China center for type culture Collection with the deposit number CCTCC NO: M2021265 or CCTCC NO: M2021266. Wherein, the CCTCC NO: M2021265 is GS115/4proROL-Mrcp19k-Sso 2-Bmh-Ssa 4-VHb strain, and the CCTCC NO: M2021266 is GS115/4proROL-Mrcp20k-Sso 2-Bmh-Ssa 4-VHb strain.

The invention also provides a method for expressing the viscose protein by using the yeast engineering bacteria, which is characterized in that when the bacterial liquid is cultured, the initial pH of the BMGY culture medium is =7.0, the inoculum size is 3%, the methanol addition amount is 1.5%, the induction temperature is 25 ℃, and the induction time is 96h.

In some embodiments, the above method uses the engineered yeast strain described above in the culture of the bacterial solution.

In some embodiments, the above method, comprises the steps of:

(1) Transforming pichia competent cells by the expression vector or the vector combination of claim 1 or 2 to obtain a yeast recombinant genetic engineering strain containing the expression vector or the vector combination;

wherein, the expression vector or the vector combination can be linearized by SalI or BamHI sites to facilitate integration; the pichia competent cells can be derived from the GS115 strain; after the competent cells are transformed, YPDS solid plates of MD or corresponding antibiotics can be used for screening to obtain the yeast recombinant genetic engineering bacteria containing the expression vector or the vector combination

(2) Inoculating the yeast recombinant genetic engineering bacteria obtained in the step (1) into 5mL YPD liquid culture medium, and culturing for 20h;

(3) Inoculating the engineering bacteria in a BMGY culture medium with initial pH =7.0 in an inoculation amount of 3% (v/v), and culturing at 28 ℃ for 24h;

(4) The thalli are collected by centrifugation and inoculated in 50mL BMMY culture medium, inducer absolute methanol is added every 24h until the final volume concentration of methanol is 1.5% (v/v), and the mixture is subjected to induced expression for 96h at 25 ℃ on a shaker at 200 rpm/min.

Wherein MD, YPD, BMGY and BMMY are all culture media commonly used in yeast culture, and other data can be referred to for specific formula.

Compared with the prior art, the invention has the advantages that: the efficient expression of the barnacle viscose proteins cp19k and cp20k by using pichia pastoris has important application value, but no data disclose a specific expression method. The expression efficiency of other barnacle viscose protein expressed data is low, and the requirement of large-scale production and application cannot be met. Based on a pichia pastoris expression system, the invention optimizes parameters such as initial pH value, inoculation quantity, methanol addition quantity, induction temperature, induction time and the like of a culture medium of the bacterial liquid by technical means of increasing copy number, fusing lipase, increasing auxiliary factors and the like, improves the expression quantity of the mucin to gram level, and meets the requirements of large-scale production and application. If the method is combined with the later use of a large-scale fermentation tank, the expression amount can be further improved.

Drawings

FIG. 1 vector diagram of pAO α N-Mrcp19 k.

FIG. 2 vector diagram of pAO α N-Mrcp20 k.

FIG. 3 SDS-PGAE results of Mrcp recombinant protein. A: mrcp19k; b: mrcp20k.

FIG. 4 shows liquid chromatography tandem mass spectrometry detection of the molecular weight of the recombinant Mrcp19k monomer.

FIG. 5 shows that the liquid chromatography tandem mass spectrometry is adopted to detect the molecular weight of the recombinant Mrcp19k dimer.

FIG. 6 vector diagram of pAO α N-4Mrcp19 k.

FIG. 7 vector diagram pAO α N-4Mrcp20 k.

FIG. 8 SDS-PGAE results of proROL-Mrcp recombinant protein. A: proROL-Mrcp19k; b: proROL-Mrcp20k. .

FIG. 9 vector diagram of pAO α N-4proROL-Mrcp19 k. The English letters of the elements and the meanings of the abbreviations are listed as follows:

AOX1 promoter

Alpha-factor secretion signal

proROL-Mrcp19k proROL-Mrcp19k fusion protein gene

AOX1 terminator

PpHIS4 yeast selection marker

AOX1 'fragment AOX1' fragment

AmpR promoter Amp gene promoter

AmpR Amp antibiotic resistance

FIG. 10 shows the vector map of pAO α N-4proROL-Mrcp20 k. The English letters of the elements and the meanings of the abbreviations are listed as follows:

AOX1 promoter

Alpha-factor secretion signal

proROL-Mrcp20k proROL-Mrcp20k fusion protein gene

AOX1 terminator

PpHIS4 yeast selection marker

AOX1 'fragment AOX1' fragment

AmpR promoter Amp gene promoter

AmpR Amp antibiotic resistance

FIG. 11 is a map of the pPICZ-Ssa4-VHb vector. The English letters of the elements and the meanings of the abbreviations are listed as follows:

AOX1 promoter

Ssa4 Ssa4 cofactor gene

Myc Myc protein tag

6 XHis protein tag

AOX1 terminator

VHb VHb cofactor genes

TEF1 promoter

EM7 promoter

Zeocin resistance to Zeocin antibiotics

CYC1 terminator

ori ori replicons

FIG. 12 depicts vector diagrams of pPIC3.5k-Sso 2-Bmh. The English letters of the elements and the meanings of the abbreviations are listed as follows:

AOX1 promoter

Sso2 Sso2 cofactor gene

AOX1 terminator

Bmh2 Bmh cofactor gene

PpHIS4 yeast selection marker

KanR KanR antibiotic resistance

AOX1 'fragment AOX1' fragment

bom TEF1 promoter

ori ori replicon

AmpR promoter Amp gene promoter

AmpR Amp antibiotic resistance

FIG. 13 shows the results of the optimized parameters of culture conditions for mucin-expressing strains. A to E: GS115/4-Mrcp20k strain test results; F-G: GS115/4proROL-Mrcp20k-Sso 2-Bmh-Ssa 4-VHb strain test results. A. F: testing initial pH value parameters of a bacterial liquid culture medium; B. g: testing inoculation quantity parameters; C. h: testing the methanol addition quantity parameter; D. i: testing induction time parameters; E. j: and (5) testing induction temperature parameters. The horizontal axis represents the parameter set value, and the vertical axis represents the protein concentration (unit: mg/L).

Detailed Description

The following definitions and methods are provided to better define the present application and to guide those of ordinary skill in the art in the practice of the present application. Unless otherwise indicated, terms are to be understood in accordance with their ordinary usage by those of ordinary skill in the relevant art. All patent documents, academic papers, industry standards and other publications, etc., cited herein are incorporated by reference in their entirety.

Those skilled in the art will readily recognize that advances in the field of molecular biology, such as site-specific and random mutagenesis, polymerase chain reaction methods, and protein engineering techniques, provide a wide range of suitable tools and procedures for engineering or engineering amino acid sequences and potentially genetic sequences of proteins of interest.

In some embodiments, changes may be made to the nucleotide sequences of the present application to make conservative amino acid substitutions. The principles and examples of conservative amino acid substitutions are further described below. In certain embodiments, substitutions that do not alter the amino acid sequence of the nucleotide sequences of the present application can be made in accordance with the disclosed yeast codon preferences, e.g., codons encoding the same amino acid sequence can be substituted with yeast preferred codons without altering the amino acid sequence encoded by the nucleotide sequence. In some embodiments, a portion of the nucleotide sequence in this application is replaced with a different codon that encodes the same amino acid sequence, such that the nucleotide sequence is not altered while the amino acid sequence encoded thereby is not altered. Conservative variants include those sequences that, due to the degeneracy of the genetic code, encode the amino acid sequence of one of the proteins of the embodiments. In some embodiments, a portion of the nucleotide sequence in the present application is replaced according to yeast preferred codons. One skilled in the art will recognize that amino acid additions and/or substitutions are generally based on the relative similarity of the amino acid side-chain substituents, e.g., hydrophobicity, charge, size, etc., of the substituents. Exemplary amino acid substituent groups having various of the foregoing properties are known to those skilled in the art and include arginine and lysine; glutamic acid and aspartic acid; serine and threonine; glutamine and asparagine; and valine, leucine and isoleucine. Guidance as to suitable amino acid substitutions that do not affect the biological activity of the Protein of interest can be found in the model of the Atlas of Protein sequences and structures (Protein Sequence and Structure Atlas) (Natl.biomed.Res.Foundation, washington, D.C.) (incorporated herein by reference). Conservative substitutions such as exchanging one amino acid for another with similar properties may be made. Identification of sequence identity includes hybridization techniques. For example, all or part of a known nucleotide sequence is used as a probe for selective hybridization to other corresponding nucleotide sequences present in a population of cloned genomic DNA fragments or cDNA fragments (i.e., a genomic library or cDNA library) from a selected organism. The hybridization probes may be genomic DNA fragments, cDNA fragments, RNA fragments, or other oligonucleotides, and may be labeled with a detectable group such as 32P or other detectable marker. Thus, for example, hybridization probes can be prepared by labeling synthetic oligonucleotides based on the sequence of the embodiment. Methods for preparing hybridization probes and constructing cDNA and genomic libraries are generally known in the art. Hybridization of the sequences may be performed under stringent conditions. As used herein, the term "stringent conditions" or "stringent hybridization conditions" refers to conditions under which a probe will hybridize to its target sequence to a detectably greater degree (e.g., at least 2-fold, 5-fold, or 10-fold over background) than to other sequences. Stringent conditions are sequence-dependent and will be different in different circumstances. By controlling the stringency of hybridization and/or the washing conditions, target sequences can be identified that are 100% complementary to the probes (homologous probe method). Alternatively, stringency conditions can be adjusted to allow some sequence mismatches in order to detect lower similarity (heterologous probe methods). Typically, probes are less than about 1000 or 500 nucleotides in length. Typically, stringent conditions are conditions in which the salt concentration is less than about 1.5M Na ion, typically about 0.01M to 1.0M Na ion concentration (or other salt) at pH 7.0 to 8.3, and the temperature conditions are: when used with short probes (e.g., 10 to 50 nucleotides), at least about 30 ℃; when used with long probes (e.g., greater than 50 nucleotides), at least about 60 ℃. Stringent conditions may also be achieved by the addition of destabilizing agents such as formamide. Exemplary low stringency conditions include hybridization using 30% to 35% formamide buffer, 1M NaCl, 1% sds (sodium dodecyl sulfate) at 37 ℃, washing in 1 x to 2 x SSC (20 x SSC =3.0M NaCl/0.3M trisodium citrate) at 50 ℃ to 55 ℃. Exemplary moderately stringent conditions include hybridization in 40% to 45% formamide, 1.0M NaCl, 1% SDS at 37 ℃ and washing in 0.5X to 1X SSC at 55 ℃ to 60 ℃. Exemplary high stringency conditions include hybridization in 50% formamide, 1M NaCl, 1% SDS at 37 ℃, and a final wash in 0.1 XSSC at 60 ℃ to 65 ℃ for at least about 20min. Optionally, the wash buffer may comprise about 0.1% to about 1% sds. The duration of hybridization is generally less than about 24 hours, usually from about 4 hours to about 12 hours. Specificity is usually dependent on the post-hybridization wash, the key factors being the ionic strength and temperature of the final wash solution. The Tm (thermodynamic melting point) of a DNA-DNA hybrid can be approximated by the formula of Meinkoth and Wahl (1984) anal. Biochem.138: 267-284: tm =81.5 ℃ +16.6 (logM) +0.41 (% GC) -0.61 (% formamide) -500/L; where M is the molar concentration of monovalent cations,% GC is the percentage of guanosine and cytosine nucleotides in the DNA,% formamide is the percentage formamide of the hybridization solution, and L is the base pair length of the hybrid. The Tm is the temperature (under defined ionic strength and pH) at which 50% of a complementary target sequence hybridizes to a perfectly matched probe. Washing is typically performed at least until equilibrium is reached and a low background level of hybridization is reached, such as for 2h, 1h or 30min. Decrease Tm by about 1 ℃ per 1% mismatch; thus, tm, hybridization and/or wash conditions can be adjusted to hybridize to sequences of desired identity. For example, if a sequence with > 90% identity is desired, the Tm can be lowered by 10 ℃. Typically, stringent conditions are selected to be about 5 ℃ lower than the Tm for the specific sequence and its complement under defined ionic strength and pH. However, under very stringent conditions, hybridization and/or washing can be performed at 4 ℃ below the Tm; hybridization and/or washing may be performed at 6 ℃ below the Tm under moderately stringent conditions; under low stringency conditions, hybridization and/or washing can be performed at 11 ℃ below the Tm.

The following examples are intended to illustrate the invention, but are not intended to limit the scope of the invention. Modifications or substitutions to methods, steps or conditions of the present invention may be made without departing from the spirit and substance of the invention and are intended to be included within the scope of the present application. Unless otherwise indicated, the examples follow conventional experimental conditions, such as those set forth in Sambrook et al, molecular cloning, A laboratory Manual (Sambrook J & Russell DW, molecular cloning: a laboratory Manual, 2001), or as recommended by the manufacturer's instructions. Unless otherwise specified, all chemical reagents used in the examples are conventional commercially available reagents, and the technical means used in the examples are conventional means well known to those skilled in the art.

Example 1 Primary expression of barnacle mucilaginous proteins

The protein Mrcp19k (GenBank: BAE94409.1 amino acids 26-198) and Mrcp20k (GenBank: BAB18762.1 amino acids 20-202) are selected as target proteins expressed by pichia pastoris. Nucleotide sequences encoding the Mrcp19k and Mrcp20k proteins were designed, codon optimized and 6 histidine tag sequences added. Mrcp19k and Mrcp20k nucleic acid molecules were synthesized artificially. Specific nucleic acid sequences for Mrcp19k and Mrcp20k are shown in SEQ ID NO.7 and SEQ ID NO.8.

The expression vector is pAO alpha N. The vector is transformed from a common commercial yeast expression vector pAO815, and a secretion signal peptide alpha-factor is added, so that extracellular secretion expression of protein can be carried out; and meanwhile, notI endonuclease sites are added, ecoRI/NotI double enzyme cutting sites are formed at two ends of the target gene, and different exogenous genes are conveniently introduced. The carrier contains a screening marker HIS4, which is convenient for multi-strategy integration to improve the protein expression quantity; introducing affinity tags of 6 histidines at the carboxyl terminal of the protein, and purifying by using a nickel affinity column; the vector resistance was AmpR.

The synthesized Mrcp19k and Mrcp20k genes are constructed on a pAOaN skeleton vector from a pPICZ alpha A vector through enzyme cutting sites EcoR I and Not I to obtain expression vectors pAOaN-Mrcp 19k and pAOaN-Mrcp 20k (vector maps are respectively shown in the attached figures 1 and 2).

And carrying out single enzyme digestion on a series of successfully constructed vectors by Sal I, transforming a Pichia pastoris GS115 strain, and screening by using an MD solid plate. Single colonies of the recombinant genetically engineered bacteria on the plate were inoculated into 5mL of YPD liquid medium and cultured for 20 hours. The recombinant strain was inoculated in 50mL of BMGY medium (initial pH 7.0) at an inoculum size of 2% (v/v) and cultured for 24 hours. The inducer methanol was supplemented every 24h until the final methanol volume concentration was 1.5% (v/v), and induction of expression 96h was complete. All the recombinant bacteria are put in a shaking table at 28 ℃ and 200rpm/min for shaking flask fermentation. SDS-PAGE electrophoresis detection was performed using 15% pre-gel and the corresponding running buffer. Total protein concentration in the supernatant of the fermentation broth was determined by Bradford method, see reagent instructions (PA 102, TIANGEN). Identification of protein peptide fragments was entrusted to Suzhou Putai Biotechnology Ltd. The molecular weight of protein was determined by Bekindskin Betay Biotech Co.

And (3) detecting the SDS-PGAE result of the recombinant Mrcp19k protein: electrophoresis results show that dispersed protein expression bands exist in the molecular weight range of 33-45kDa, peptide fragment identification is carried out by Nano-LC-ESI-MS/MS protein mass spectrum, and partial peptide fragments can be matched with the amino acid sequence of the protein, so that the dispersed protein is recombinant Mrcp19k protein (shown in figure 3-A). However, the actual molecular weight of the protein is about 2 times greater than the theoretical molecular weight of 17.8 kDa. Further, the molecular weight of the recombinant Mrcp19k is detected by liquid chromatography tandem mass spectrometry, and as can be seen from the deconvolution molecular weights, the recombinant Mrcp19k exists in the form of a monomer and a dimer, respectively, the molecular weight of the monomer is 17.855kDa (shown in figure 4), and the molecular weight of the dimer is 35.708kDa (shown in figure 5). Therefore, the successful expression of the recombinant Mrcp19k protein in Pichia pastoris is demonstrated.

And (3) detecting the result of SDS-PGAE of the recombinant Mrcp20k protein: the electrophoresis result shows that the protein expression band is at the molecular weight of about 30kDa, which is about 9kDa larger than the theoretical molecular weight of 21.2 kDa. Peptide fragment identification is carried out by Nano-LC-ESI-MS/MS protein spectrum, and partial peptide fragment can be matched with the amino acid sequence of the protein, which indicates that the protein is recombinant Mrcp20k protein (figure 3-B).

Preliminary secretory expression results of Mrcp19k and Mrcp20k in a Pichia pastoris system show that the initial expression of the two proteins is successful, but the expression amount is low, wherein the total protein concentration secreted by the Mrcp19k is 48mg/L, and the total protein concentration secreted by the Mrcp20k is 62mg/L.

Example 2 increasing expression levels by increasing copy number of target protein

Increasing the copy number of the target gene is often effective in increasing the expression level of the target protein. The invention adopts a biological brick method to construct Mrcp19k and Mrcp20k expression vectors with different copies so as to test the promotion effect of multiple copies on the expression quantity and screen the optimal copy number. Multicopy vector construction methods reference: china university of science and technology, a method for constructing yeast multicopy expression vector, CN201610172061.5[ P ].2016-06-07. The endonuclease is selected from isocaudarner BamH I and Bgl II.

The target protein is expressed by using constructed pAO alpha N-nMrcp19k and pAO alpha N-nMrcp20k (N =2,3,4) multi-copy vectors. The expression system is referred to example 1. Protein concentration measurement results show that the total protein concentrations secreted by the Mrcp19k with 2 copies, 3 copies and 4 copies are 90mg/L, 119mg/L and 151mg/L respectively; the total protein concentration secreted by Mrcp20k at 2 copies, 3 copies and 4 copies was 103mg/L, 150mg/L and 172mg/L, respectively. Therefore, the protein expression efficiency of the 4 copies of Mrcp19k or Mrcp20k is the highest. Since the 5-copy vector fragment is too large to be successfully constructed, 4 copies is the optimal strategy to integrate vector construction and protein concentration. The expression vector maps of pAO alpha N-4Mrcp19K and pAO alpha N-4Mrcp20K are shown in the attached figures 6 and 7, and the vector sequences are shown in SEQ ID NO.1 and SEQ ID NO. 2.

Example 3 expression of Rhizopus oryzae Lipase by fusion

Rhizopus Oryzae Lipase (ROL) has a typical intramolecular chaperone structure and can promote the expression and secretion of target protein.

Barnacle mucilaginous proteins Mrcp19k and Mrcp20k were fused to ROL, respectively. Wherein ROL is at the N-terminus, mrcp protein (containing a histidine tag) is at the C-terminus, and a common flexible linker peptide (G) is placed in between ₄ S) ₃ The connection is made.

The sequences of proROL-Mrcp19k and proROL-Mrcp20k are artificially synthesized, and the specific sequences are shown in SEQ ID NO.9 and SEQ ID NO.10.

The fragment of proROL-Mrcp19k and proROL-Mrcp20k is connected to the pAOaN framework vector through EcoRI and NotI to obtain the pAOaN-proROL-Mrcp 19k and pAOaN-proROL-Mrcp 20k recombinant vectors. Meanwhile, pAO α N-npROL-Mrcp 19k and pAO α N-npROL-Mrcp 20k (N =2,3,4) multicopy vectors were constructed according to the method described in example 2.

The target protein is expressed by using the vector containing different copies of proROL-Mrcp19k and proROL-Mrcp20k fragments. The expression system is referred to example 1.

The SDS-PGAE detection result of the recombinant proROL-Mrcp19k protein shows that the expression condition of the proROL-Mrcp19k protein is similar to the expression condition of the recombinant Mrcp19k protein, dispersed electrophoresis bands exist in the range of 60-75kDa, the molecular weight of electrophoresis detection is larger than the predicted molecular weight (58.2 kDa) (shown in figure 8-A), and the mass spectrum detection result shows that the protein is successfully expressed.

The results of SDS-PGAE detection of the recombinant proROL-Mrcp20k protein revealed that the molecular weight of the proROL-Mrcp20k protein was about 60kDa, which is substantially identical to the predicted molecular weight of 61.2kDa (FIG. 8-B). Meanwhile, the mass spectrum detection result shows that the protein band is the proROL-Mrcp20k protein.

Protein concentration measurement results show that the total protein concentrations secreted by 1 copy, 2 copies, 3 copies and 4 copies of proROL-Mrcp19k are 146mg/L, 202mg/L, 278mg/L and 358mg/L respectively; the total protein concentration secreted by 1 copy, 2 copies, 3 copies and 4 copies of proROL-Mrcp20k is 210mg/L, 303mg/L, 372mg/L and 410 mg/L respectively. Therefore, after the rhizopus oryzae lipase is fused, the total expression quantity of the protein at different copy numbers is obviously improved, and the protein expression efficiency of 4 copies of Mrcp19k or Mrcp20k is still the highest in all the fused rhizopus oryzae lipase vectors. The vectors pAO alpha N-4proROL-Mrcp19k and pAO alpha N-4proROL-Mrcp20k are shown in attached figure 9 and attached figure 10, and the sequences of the vectors are shown in SEQ ID NO.3 and SEQ ID NO. 4.

Example 4 Co-expression in combination with cofactors

Research data already shows that when rhizopus oryzae lipase is expressed in pichia pastoris and co-expressed with Bmh, sso2, ssa4 and VHb which are 4 auxiliary proteins, the total protein content secreted from the extracellular can reach 7.2g/L (Jiao Liangcheng. Pichia pastoris molecular operation method improvement and application thereof in rhizopus oryzae lipase high-efficiency expression [ D ]. Science and technology university in china, 2019). In the invention, because the viscose protein and the rhizopus oryzae lipase are expressed in a fusion manner, the expression quantity of the fusion protein can be further improved by adopting a strategy of promoting the expression of the rhizopus oryzae lipase.

The invention is intended to test the effect of increasing combinations of cofactors for Ssa4-VHb and Sso2-Bmh on the expression efficiency of Mrcp19k and Mrcp20k proteins.

Constructing pPICZ-Ssa4-VHb and pPIC3.5k-Sso2-Bmh vectors, wherein the construction method refers to the following steps: jiao Liangcheng pichia pastoris molecular manipulation improvements and their use in rhizopus oryzae lipase high efficiency expression [ D ]. University of science and technology in china, 2019. The pPICZ-Ssa4-VHb and pPIC3.5k-Sso2-Bmh vectors are shown in the attached figures 11 and 12, and the specific sequences are shown in SEQ ID NO.5 and SEQ ID NO. 6. And (3) screening the Mrcp19k and Mrcp20k 4 copy fusion protein recombinant strains with the highest expression quantity as competence, and performing linearization treatment on the two types of recombinant plasmids and sequentially performing electrotransformation into competent cells. First, pPIC3.5k-Sso2-Bmh plasmid was transformed, and recombinant bacteria were screened using YPDS plates containing 1mg/mL geneticin G418. The strain with high expression amount of target protein is used as a competent cell, then pPICZ-Ssa4-VHb plasmid is transformed, and the recombinant strain is coated on a YPDS resistant plate containing 100 mu g/mL Zeocin for screening.

Protein concentration measurement results show that after Sso2-Bmh2 cofactor is increased, the expression amounts of Mrcp19k and Mrcp20k are 432mg/L and 499mg/L respectively; after further addition of Ssa4-VHb cofactor, the expression levels of Mrcp19k and Mrcp20k were 503mg/L and 621mg/L, respectively. Therefore, after increasing the Ssa4-VHb-Sso2-Bmh cofactor, the expression level of Mrcp19k and Mrcp20k can be further greatly improved.

Therefore, the fusion protein strain GS115/4proROL-Mrcp19k-Sso 2-Bmh-Ssa 4-VHb and GS115/4proROL-Mrcp20k-Sso 2-Bmh-Ssa 4-VHb are the optimal strain, and the protein expression amounts of Mrcp19k and Mrcp20k can reach 503mg/L and 621mg/L respectively. GS115/4proROL-Mrcp19k-Sso 2-Bmh-Ssa 4-VHb strain has been deposited in China center for type culture Collection with the deposit number CCTCC NO: M2021265; GS115/4proROL-Mrcp20k-Sso 2-Bmh-Ssa 4-VHb strain has been deposited with the deposit number CCTCC NO: M2021266 in the China center for type culture Collection.

The specific expression vector optimization parameters and the corresponding expression results are summarized in Table 1.

TABLE 1 summary of optimized parameters and corresponding expression levels for expression vectors for mucin

Expression values are expressed as mean. + -. Standard deviation.

The proROL-Mrcp19k and proROL-Mrcp20k proteins which are successfully expressed can be directly applied to industrial production, and protease cleavage sites can be added in advance to cleave the proROL proteins at the later stage by using protease.

Example 5 optimization of expression conditions and increase of expression amount

The culture condition of the strain is also an important factor influencing the protein expression quantity in a pichia pastoris expression system, and in the culture condition of the strain, the initial pH value, the inoculation quantity, the methanol addition quantity, the induction time, the induction temperature and other parameters of a strain liquid culture medium are the most important influencing factors.

According to the study on the culture conditions of the optimal strain for expressing rhizopus oryzae lipase by a pichia pastoris system (Jiao Liangcheng. Improvement of a pichia pastoris molecular operation method and application thereof in efficient expression of rhizopus oryzae lipase [ D ]. University of science and technology in China, 2019), the invention firstly sets the standard culture conditions of strain culture as follows: initial pH of the culture medium =7.0, inoculum size 2%, methanol addition 1.5%, induction time 96h, induction temperature 28 ℃. Then, test optimization is performed on each parameter one by one, and the set value of each parameter during testing is shown in table 2. The specific strain culture and inducible expression procedures are as described in example 1.

TABLE 2 optimization parameters of culture conditions for expression strains of viscose protein

Two strains GS115/4Mrcp20k and GS115/4proROL-Mrcp20k-Sso 2-Bmh-Ssa 4-VHb were used as test strains, and optimum parameter conditions were determined when each parameter was changed one by one under the conditions of the culture of the reference bacterial liquid for the test, and the test results are shown in FIG. 13.

According to the results, when the initial pH of the culture medium is =7.0, the inoculation amount is 3%, the addition amount of methanol is 1.5%, the induction time is 96h, and the induction temperature is 25 ℃, the protein concentration of protein Mrcp20k expressed by two strains GS115/4-Mrcp20k and GS115/4proROL-Mrcp20k-Sso 2-Bmh-Ssa 4-VHb is the highest, and can respectively reach 182mg/L and 629mg/L.

Although the invention has been described in detail with respect to the general description and the specific embodiments thereof, it will be apparent to those skilled in the art that modifications and improvements can be made based on the invention. Accordingly, such modifications and improvements are intended to be within the scope of the invention as claimed.

Sequence listing

<110> university of science and technology in Huazhong

<120> yeast engineering bacterium for efficiently expressing barnacle viscose protein and preparation method thereof

<130> 1

<160> 10

<170> SIPOSequenceListing 1.0

<210> 1

<211> 14820

<212> DNA

<213> unknown (Artificial Synthesis)

<400> 1

agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 60

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 120

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 180

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 240

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 300

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 360

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 420

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 480

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 540

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 600

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 660

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 720

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 780

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 840

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 900

caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 960

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 1020

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 1080

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 1140

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 1200

tgaagctgaa ttcgttccac ctccatgtga tttgggtatt gcttctaagg ttaaacaaaa 1260

gggtgttact ggtggtggtg cttctgtttc tactacttct gctactcaag gttctggtac 1320

tactaactgt gttactagaa ctcctaattc tgttgaaaag aaaaacgttg ctggtaatac 1380

tggtgttact gctacttctg tttctgctgg agatggtgct tttggtaact tggctgctgc 1440

tttgactttg gttgaagata ctgaggatgg tttgggtgtt aaaactaaga acggtggtaa 1500

aggtttctct gagggtactg ctgctatttc tcaaactgct ggtgctaatg gtggtgctac 1560

tgttaagaaa gcaaagttgg atttgttgac tgatggtgaa gatttgttcg atactaagaa 1620

agttgagaag ggtactgtta cttcttcttc ttctcatcaa ggttctggtg ctggagattc 1680

tatcttcgaa atcttgaacg aagctgagtc taagattaag aaatctggag atcatcacca 1740

tcaccatcac taagcggccg cgtttgtagc cttagacatg actgttcctc agttcaagtt 1800

gggcacttac gagaagaccg gtcttgctag attctaatca agaggatgtc agaatgccat 1860

ttgcctgaga gatgcaggct tcatttttga tactttttta tttgtaacct atatagtata 1920

ggattttttt tgtcattttg tttcttctcg tacgagcttg ctcctgatca gcctatctcg 1980

cagctgatga atatcttgtg gtaggggttt gggaaaatca ttcgagtttg atgtttttct 2040

tggtatttcc cactcctctt cagagtacag aagattaagt gagaccttcg tttgtgcgga 2100

tctaacatcc aaagacgaaa ggttgaatga aacctttttg ccatccgaca tccacaggtc 2160

cattctcaca cataagtgcc aaacgcaaca ggaggggata cactagcagc agaccgttgc 2220

aaacgcagga cctccactcc tcttctcctc aacacccact tttgccatcg aaaaaccagc 2280

ccagttattg ggcttgattg gagctcgctc attccaattc cttctattag gctactaaca 2340

ccatgacttt attagcctgt ctatcctggc ccccctggcg aggttcatgt ttgtttattt 2400

ccgaatgcaa caagctccgc attacacccg aacatcactc cagatgaggg ctttctgagt 2460

gtggggtcaa atagtttcat gttccccaaa tggcccaaaa ctgacagttt aaacgctgtc 2520

ttggaaccta atatgacaaa agcgtgatct catccaagat gaactaagtt tggttcgttg 2580

aaatgctaac ggccagttgg tcaaaaagaa acttccaaaa gtcggcatac cgtttgtctt 2640

gtttggtatt gattgacgaa tgctcaaaaa taatctcatt aatgcttagc gcagtctctc 2700

tatcgcttct gaaccccggt gcacctgtgc cgaaacgcaa atggggaaac acccgctttt 2760

tggatgatta tgcattgtct ccacattgta tgcttccaag attctggtgg gaatactgct 2820

gatagcctaa cgttcatgat caaaatttaa ctgttctaac ccctacttga cagcaatata 2880

taaacagaag gaagctgccc tgtcttaaac cttttttttt atcatcatta ttagcttact 2940

ttcataattg cgactggttc caattgacaa gcttttgatt ttaacgactt ttaacgacaa 3000

cttgagaaga tcaaaaaaca actaattatt cgaaacgatg agatttcctt caatttttac 3060

tgctgtttta ttcgcagcat cctccgcatt agctgctcca gtcaacacta caacagaaga 3120

tgaaacggca caaattccgg ctgaagctgt catcggttac tcagatttag aaggggattt 3180

cgatgttgct gttttgccat tttccaacag cacaaataac gggttattgt ttataaatac 3240

tactattgcc agcattgctg ctaaagaaga aggggtatct ctcgagaaaa gagaggctga 3300

agctgaattc gttccacctc catgtgattt gggtattgct tctaaggtta aacaaaaggg 3360

tgttactggt ggtggtgctt ctgtttctac tacttctgct actcaaggtt ctggtactac 3420

taactgtgtt actagaactc ctaattctgt tgaaaagaaa aacgttgctg gtaatactgg 3480

tgttactgct acttctgttt ctgctggaga tggtgctttt ggtaacttgg ctgctgcttt 3540

gactttggtt gaagatactg aggatggttt gggtgttaaa actaagaacg gtggtaaagg 3600

tttctctgag ggtactgctg ctatttctca aactgctggt gctaatggtg gtgctactgt 3660

taagaaagca aagttggatt tgttgactga tggtgaagat ttgttcgata ctaagaaagt 3720

tgagaagggt actgttactt cttcttcttc tcatcaaggt tctggtgctg gagattctat 3780

cttcgaaatc ttgaacgaag ctgagtctaa gattaagaaa tctggagatc atcaccatca 3840

ccatcactaa gcggccgcgt ttgtagcctt agacatgact gttcctcagt tcaagttggg 3900

cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg 3960

cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga 4020

ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag 4080

ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg 4140

tatttcccac tcctcttcag agtacagaag attaagtgag accttcgttt gtgcggatct 4200

aacatccaaa gacgaaaggt tgaatgaaac ctttttgcca tccgacatcc acaggtccat 4260

tctcacacat aagtgccaaa cgcaacagga ggggatacac tagcagcaga ccgttgcaaa 4320

cgcaggacct ccactcctct tctcctcaac acccactttt gccatcgaaa aaccagccca 4380

gttattgggc ttgattggag ctcgctcatt ccaattcctt ctattaggct actaacacca 4440

tgactttatt agcctgtcta tcctggcccc cctggcgagg ttcatgtttg tttatttccg 4500

aatgcaacaa gctccgcatt acacccgaac atcactccag atgagggctt tctgagtgtg 4560

gggtcaaata gtttcatgtt ccccaaatgg cccaaaactg acagtttaaa cgctgtcttg 4620

gaacctaata tgacaaaagc gtgatctcat ccaagatgaa ctaagtttgg ttcgttgaaa 4680

tgctaacggc cagttggtca aaaagaaact tccaaaagtc ggcataccgt ttgtcttgtt 4740

tggtattgat tgacgaatgc tcaaaaataa tctcattaat gcttagcgca gtctctctat 4800

cgcttctgaa ccccggtgca cctgtgccga aacgcaaatg gggaaacacc cgctttttgg 4860

atgattatgc attgtctcca cattgtatgc ttccaagatt ctggtgggaa tactgctgat 4920

agcctaacgt tcatgatcaa aatttaactg ttctaacccc tacttgacag caatatataa 4980

acagaaggaa gctgccctgt cttaaacctt tttttttatc atcattatta gcttactttc 5040

ataattgcga ctggttccaa ttgacaagct tttgatttta acgactttta acgacaactt 5100

gagaagatca aaaaacaact aattattcga aacgatgaga tttccttcaa tttttactgc 5160

tgttttattc gcagcatcct ccgcattagc tgctccagtc aacactacaa cagaagatga 5220

aacggcacaa attccggctg aagctgtcat cggttactca gatttagaag gggatttcga 5280

tgttgctgtt ttgccatttt ccaacagcac aaataacggg ttattgttta taaatactac 5340

tattgccagc attgctgcta aagaagaagg ggtatctctc gagaaaagag aggctgaagc 5400

tgaattcgtt ccacctccat gtgatttggg tattgcttct aaggttaaac aaaagggtgt 5460

tactggtggt ggtgcttctg tttctactac ttctgctact caaggttctg gtactactaa 5520

ctgtgttact agaactccta attctgttga aaagaaaaac gttgctggta atactggtgt 5580

tactgctact tctgtttctg ctggagatgg tgcttttggt aacttggctg ctgctttgac 5640

tttggttgaa gatactgagg atggtttggg tgttaaaact aagaacggtg gtaaaggttt 5700

ctctgagggt actgctgcta tttctcaaac tgctggtgct aatggtggtg ctactgttaa 5760

gaaagcaaag ttggatttgt tgactgatgg tgaagatttg ttcgatacta agaaagttga 5820

gaagggtact gttacttctt cttcttctca tcaaggttct ggtgctggag attctatctt 5880

cgaaatcttg aacgaagctg agtctaagat taagaaatct ggagatcatc accatcacca 5940

tcactaagcg gccgcgtttg tagccttaga catgactgtt cctcagttca agttgggcac 6000

ttacgagaag accggtcttg ctagattcta atcaagagga tgtcagaatg ccatttgcct 6060

gagagatgca ggcttcattt ttgatacttt tttatttgta acctatatag tataggattt 6120

tttttgtcat tttgtttctt ctcgtacgag cttgctcctg atcagcctat ctcgcagctg 6180

atgaatatct tgtggtaggg gtttgggaaa atcattcgag tttgatgttt ttcttggtat 6240

ttcccactcc tcttcagagt acagaagatt aagtgagacc ttcgtttgtg cggatctaac 6300

atccaaagac gaaaggttga atgaaacctt tttgccatcc gacatccaca ggtccattct 6360

cacacataag tgccaaacgc aacaggaggg gatacactag cagcagaccg ttgcaaacgc 6420

aggacctcca ctcctcttct cctcaacacc cacttttgcc atcgaaaaac cagcccagtt 6480

attgggcttg attggagctc gctcattcca attccttcta ttaggctact aacaccatga 6540

ctttattagc ctgtctatcc tggcccccct ggcgaggttc atgtttgttt atttccgaat 6600

gcaacaagct ccgcattaca cccgaacatc actccagatg agggctttct gagtgtgggg 6660

tcaaatagtt tcatgttccc caaatggccc aaaactgaca gtttaaacgc tgtcttggaa 6720

cctaatatga caaaagcgtg atctcatcca agatgaacta agtttggttc gttgaaatgc 6780

taacggccag ttggtcaaaa agaaacttcc aaaagtcggc ataccgtttg tcttgtttgg 6840

tattgattga cgaatgctca aaaataatct cattaatgct tagcgcagtc tctctatcgc 6900

ttctgaaccc cggtgcacct gtgccgaaac gcaaatgggg aaacacccgc tttttggatg 6960

attatgcatt gtctccacat tgtatgcttc caagattctg gtgggaatac tgctgatagc 7020

ctaacgttca tgatcaaaat ttaactgttc taacccctac ttgacagcaa tatataaaca 7080

gaaggaagct gccctgtctt aaaccttttt ttttatcatc attattagct tactttcata 7140

attgcgactg gttccaattg acaagctttt gattttaacg acttttaacg acaacttgag 7200

aagatcaaaa aacaactaat tattcgaaac gatgagattt ccttcaattt ttactgctgt 7260

tttattcgca gcatcctccg cattagctgc tccagtcaac actacaacag aagatgaaac 7320

ggcacaaatt ccggctgaag ctgtcatcgg ttactcagat ttagaagggg atttcgatgt 7380

tgctgttttg ccattttcca acagcacaaa taacgggtta ttgtttataa atactactat 7440

tgccagcatt gctgctaaag aagaaggggt atctctcgag aaaagagagg ctgaagctga 7500

attcgttcca cctccatgtg atttgggtat tgcttctaag gttaaacaaa agggtgttac 7560

tggtggtggt gcttctgttt ctactacttc tgctactcaa ggttctggta ctactaactg 7620

tgttactaga actcctaatt ctgttgaaaa gaaaaacgtt gctggtaata ctggtgttac 7680

tgctacttct gtttctgctg gagatggtgc ttttggtaac ttggctgctg ctttgacttt 7740

ggttgaagat actgaggatg gtttgggtgt taaaactaag aacggtggta aaggtttctc 7800

tgagggtact gctgctattt ctcaaactgc tggtgctaat ggtggtgcta ctgttaagaa 7860

agcaaagttg gatttgttga ctgatggtga agatttgttc gatactaaga aagttgagaa 7920

gggtactgtt acttcttctt cttctcatca aggttctggt gctggagatt ctatcttcga 7980

aatcttgaac gaagctgagt ctaagattaa gaaatctgga gatcatcacc atcaccatca 8040

ctaagcggcc gcgtttgtag ccttagacat gactgttcct cagttcaagt tgggcactta 8100

cgagaagacc ggtcttgcta gattctaatc aagaggatgt cagaatgcca tttgcctgag 8160

agatgcaggc ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt 8220

ttgtcatttt gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagctgatg 8280

aatatcttgt ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc 8340

ccactcctct tcagagtaca gaagattaag tgagaccttc gtttgtgcgg atcctaatgc 8400

ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat 8460

gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg 8520

ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat 8580

ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca 8640

ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc 8700

gactacgcga tcatggcgac cacacccgtc ctgtggatct atcgaatcta aatgtaagtt 8760

aaaatctcta aataattaaa taagtcccag tttctccata cgaaccttaa cagcattgcg 8820

gtgagcatct agaccttcaa cagcagccag atccatcact gcttggccaa tatgtttcag 8880

tccctcagga gttacgtctt gtgaagtgat gaacttctgg aaggttgcag tgttaactcc 8940

gctgtattga cgggcatatc cgtacgttgg caaagtgtgg ttggtaccgg aggagtaatc 9000

tccacaactc tctggagagt aggcaccaac aaacacagat ccagcgtgtt gtacttgatc 9060

aacataagaa gaagcattct cgatttgcag gatcaagtgt tcaggagcgt actgattgga 9120

catttccaaa gcctgctcgt aggttgcaac cgatagggtt gtagagtgtg caatacactt 9180

gcgtacaatt tcaacccttg gcaactgcac agcttggttg tgaacagcat cttcaattct 9240

ggcaagctcc ttgtctgtca tatcgacagc caacagaatc acctgggaat caataccatg 9300

ttcagcttga gacagaaggt ctgaggcaac gaaatctgga tcagcgtatt tatcagcaat 9360

aactagaact tcagaaggcc cagcaggcat gtcaatacta cacagggctg atgtgtcatt 9420

ttgaaccatc atcttggcag cagtaacgaa ctggtttcct ggaccaaata ttttgtcaca 9480

cttaggaaca gtttctgttc cgtaagccat agcagctact gcctgggcgc ctcctgctag 9540

cacgatacac ttagcaccaa ccttgtgggc aacgtagatg acttctgggg taagggtacc 9600

atccttctta ggtggagatg caaaaacaat ttctttgcaa ccagcaactt tggcaggaac 9660

acccagcatc agggaagtgg aaggcagaat tgcggttcca ccaggaatat agaggccaac 9720

tttctcaata ggtcttgcaa aacgagagca gactacacca gggcaagtct caacttgcaa 9780

cgtctccgtt agttgagctt catggaattt cctgacgtta tctatagaga gatcaatggc 9840

tctcttaacg ttatctggca attgcataag ttcctctggg aaaggagctt ctaacacagg 9900

tgtcttcaaa gcgactccat caaacttggc agttagttct aaaagggctt tgtcaccatt 9960

ttgacgaaca ttgtcgacaa ttggtttgac taattccata atctgttccg ttttctggat 10020

aggacgacga agggcatctt caatttcttg tgaggaggcc ttagaaacgt caattttgca 10080

caattcaata cgaccttcag aagggacttc tttaggtttg gattcttctt taggttgttc 10140

cttggtgtat cctggcttgg catctccttt ccttctagtg acctttaggg acttcatatc 10200

caggtttctc tccacctcgt ccaacgtcac accgtacttg gcacatctaa ctaatgcaaa 10260

ataaaataag tcagcacatt cccaggctat atcttccttg gatttagctt ctgcaagttc 10320

atcagcttcc tccctaattt tagcgttcaa caaaacttcg tcgtcaaata accgtttggt 10380

ataagaacct tctggagcat tgctcttacg atcccacaag gtggcttcca tggctctaag 10440

accctttgat tggccaaaac aggaagtgcg ttccaagtga cagaaaccaa cacctgtttg 10500

ttcaaccaca aatttcaagc agtctccatc acaatccaat tcgataccca gcaacttttg 10560

agttgctcca gatgtagcac ctttatacca caaaccgtga cgacgagatt ggtagactcc 10620

agtttgtgtc cttatagcct ccggaataga ctttttggac gagtacacca ggcccaacga 10680

gtaattagaa gagtcagcca ccaaagtagt gaatagacca tcggggcggt cagtagtcaa 10740

agacgccaac aaaatttcac tgacagggaa ctttttgaca tcttcagaaa gttcgtattc 10800

agtagtcaat tgccgagcat caataatggg gattatacca gaagcaacag tggaagtcac 10860

atctaccaac tttgcggtct cagaaaaagc ataaacagtt ctactaccgc cattagtgaa 10920

acttttcaaa tcgcccagtg gagaagaaaa aggcacagcg atactagcat tagcgggcaa 10980

ggatgcaact ttatcaacca gggtcctata gataacccta gcgcctggga tcatcctttg 11040

gacaactctt tctgccaaat ctaggtccaa aatcacttca ttgataccat tattgtacaa 11100

cttgagcaag ttgtcgatca gctcctcaaa ttggtcctct gtaacggatg actcaacttg 11160

cacattaact tgaagctcag tcgattgagt gaacttgatc aggttgtgca gctggtcagc 11220

agcataggga aacacggctt ttcctaccaa actcaaggaa ttatcaaact ctgcaacact 11280

tgcgtatgca ggtagcaagg gaaatgtcat acttgaagtc ggacagtgag tgtagtcttg 11340

agaaattctg aagccgtatt tttattatca gtgagtcagt catcaggaga tcctctacgc 11400

cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc 11460

cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg 11520

cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc 11580

accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat 11640

gcaggagtcg cataagggag agcgtcgagt atctatgatt ggaagtatgg gaatggtgat 11700

acccgcattc ttcagtgtct tgaggtctcc tatcagatta tgcccaacta aagcaaccgg 11760

aggaggagat ttcatggtaa atttctctga cttttggtca tcagtagact cgaactgtga 11820

gactatctcg gttatgacag cagaaatgtc cttcttggag acagtaaatg aagtcccacc 11880

aataaagaaa tccttgttat caggaacaaa cttcttgttt cgaacttttt cggtgccttg 11940

aactataaaa tgtagagtgg atatgtcggg taggaatgga gcgggcaaat gcttaccttc 12000

tggaccttca agaggtatgt agggtttgta gatactgatg ccaacttcag tgacaacgtt 12060

gctatttcgt tcaaaccatt ccgaatccag agaaatcaaa gttgtttgtc tactattgat 12120

ccaagccagt gcggtcttga aactgacaat agtgtgctcg tgttttgagg tcatctttgt 12180

atgaataaat ctagtctttg atctaaataa tcttgacgag ccaaggcgat aaatacccaa 12240

atctaaaact cttttaaaac gttaaaagga caagtatgtc tgcctgtatt aaaccccaaa 12300

tcagctcgta gtctgatcct catcaacttg aggggcacta tcttgtttta gagaaatttg 12360

cggagatgcg atatcgagaa aaaggtacgc tgattttaaa cgtgaaattt atctcaagat 12420

ctgctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga 12480

gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc 12540

agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt 12600

gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 12660

tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc 12720

tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 12780

aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 12840

aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 12900

ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 12960

acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 13020

ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 13080

tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 13140

tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 13200

gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 13260

agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 13320

tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 13380

agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 13440

tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 13500

acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 13560

tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 13620

agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 13680

tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 13740

acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 13800

tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 13860

ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 13920

agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg 13980

tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 14040

acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 14100

agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 14160

actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 14220

tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc 14280

gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 14340

ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 14400

tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 14460

aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 14520

tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 14580

tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 14640

gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 14700

ccctttcgtc ttcaagaatt aattctcatg tttgacagct tatcatcgat aagctgactc 14760

atgttggtat tgtgaaatag acgcagatcg ggaacactga aaaataacag ttattattcg 14820

<210> 2

<211> 14940

<212> DNA

<213> unknown (Artificial Synthesis)

<400> 2

agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 60

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 120

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 180

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 240

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 300

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 360

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 420

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 480

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 540

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 600

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 660

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 720

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 780

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 840

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 900

caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 960

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 1020

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 1080

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 1140

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 1200

tgaagctgaa ttccatgaag aggatggtgt ttgtaactct aatgctccat gttaccactg 1260

tgatgctaac ggtgaaaact gttcttgtaa ctgtgaattg tttgattgtg aggctaagaa 1320

accagatggt tcttatgctc atccttgtag aagatgtgat gctaacaaca tctgtaagtg 1380

ttcttgtact gctatcccat gtaatgaaga tcatccttgt catcactgtc acgaagagga 1440

tgatggagat actcattgtc actgttcttg tgagcattct cacgatcatc acgatgatga 1500

tactcatggt gaatgtacta agaaagctcc atgttggcgt tgtgagtaca acgctgattt 1560

gaagcatgat gtttgtggtt gtgaatgttc taaattgcca tgtaatgatg aacacccttg 1620

ttatagaaag gagggtggtg ttgtttcttg tgattgtaag actatcactt gtaacgagga 1680

tcatccttgt taccactctt atgaagagga tggtgttact aagtctgatt gtgattgtga 1740

acattctcca ggtccttctg agcatcacca tcaccatcac taagcggccg cgtttgtagc 1800

cttagacatg actgttcctc agttcaagtt gggcacttac gagaagaccg gtcttgctag 1860

attctaatca agaggatgtc agaatgccat ttgcctgaga gatgcaggct tcatttttga 1920

tactttttta tttgtaacct atatagtata ggattttttt tgtcattttg tttcttctcg 1980

tacgagcttg ctcctgatca gcctatctcg cagctgatga atatcttgtg gtaggggttt 2040

gggaaaatca ttcgagtttg atgtttttct tggtatttcc cactcctctt cagagtacag 2100

aagattaagt gagaccttcg tttgtgcgga tctaacatcc aaagacgaaa ggttgaatga 2160

aacctttttg ccatccgaca tccacaggtc cattctcaca cataagtgcc aaacgcaaca 2220

ggaggggata cactagcagc agaccgttgc aaacgcagga cctccactcc tcttctcctc 2280

aacacccact tttgccatcg aaaaaccagc ccagttattg ggcttgattg gagctcgctc 2340

attccaattc cttctattag gctactaaca ccatgacttt attagcctgt ctatcctggc 2400

ccccctggcg aggttcatgt ttgtttattt ccgaatgcaa caagctccgc attacacccg 2460

aacatcactc cagatgaggg ctttctgagt gtggggtcaa atagtttcat gttccccaaa 2520

tggcccaaaa ctgacagttt aaacgctgtc ttggaaccta atatgacaaa agcgtgatct 2580

catccaagat gaactaagtt tggttcgttg aaatgctaac ggccagttgg tcaaaaagaa 2640

acttccaaaa gtcggcatac cgtttgtctt gtttggtatt gattgacgaa tgctcaaaaa 2700

taatctcatt aatgcttagc gcagtctctc tatcgcttct gaaccccggt gcacctgtgc 2760

cgaaacgcaa atggggaaac acccgctttt tggatgatta tgcattgtct ccacattgta 2820

tgcttccaag attctggtgg gaatactgct gatagcctaa cgttcatgat caaaatttaa 2880

ctgttctaac ccctacttga cagcaatata taaacagaag gaagctgccc tgtcttaaac 2940

cttttttttt atcatcatta ttagcttact ttcataattg cgactggttc caattgacaa 3000

gcttttgatt ttaacgactt ttaacgacaa cttgagaaga tcaaaaaaca actaattatt 3060

cgaaacgatg agatttcctt caatttttac tgctgtttta ttcgcagcat cctccgcatt 3120

agctgctcca gtcaacacta caacagaaga tgaaacggca caaattccgg ctgaagctgt 3180

catcggttac tcagatttag aaggggattt cgatgttgct gttttgccat tttccaacag 3240

cacaaataac gggttattgt ttataaatac tactattgcc agcattgctg ctaaagaaga 3300

aggggtatct ctcgagaaaa gagaggctga agctgaattc catgaagagg atggtgtttg 3360

taactctaat gctccatgtt accactgtga tgctaacggt gaaaactgtt cttgtaactg 3420

tgaattgttt gattgtgagg ctaagaaacc agatggttct tatgctcatc cttgtagaag 3480

atgtgatgct aacaacatct gtaagtgttc ttgtactgct atcccatgta atgaagatca 3540

tccttgtcat cactgtcacg aagaggatga tggagatact cattgtcact gttcttgtga 3600

gcattctcac gatcatcacg atgatgatac tcatggtgaa tgtactaaga aagctccatg 3660

ttggcgttgt gagtacaacg ctgatttgaa gcatgatgtt tgtggttgtg aatgttctaa 3720

attgccatgt aatgatgaac acccttgtta tagaaaggag ggtggtgttg tttcttgtga 3780

ttgtaagact atcacttgta acgaggatca tccttgttac cactcttatg aagaggatgg 3840

tgttactaag tctgattgtg attgtgaaca ttctccaggt ccttctgagc atcaccatca 3900

ccatcactaa gcggccgcgt ttgtagcctt agacatgact gttcctcagt tcaagttggg 3960

cacttacgag aagaccggtc ttgctagatt ctaatcaaga ggatgtcaga atgccatttg 4020

cctgagagat gcaggcttca tttttgatac ttttttattt gtaacctata tagtatagga 4080

ttttttttgt cattttgttt cttctcgtac gagcttgctc ctgatcagcc tatctcgcag 4140

ctgatgaata tcttgtggta ggggtttggg aaaatcattc gagtttgatg tttttcttgg 4200

tatttcccac tcctcttcag agtacagaag attaagtgag accttcgttt gtgcggatct 4260

aacatccaaa gacgaaaggt tgaatgaaac ctttttgcca tccgacatcc acaggtccat 4320

tctcacacat aagtgccaaa cgcaacagga ggggatacac tagcagcaga ccgttgcaaa 4380

cgcaggacct ccactcctct tctcctcaac acccactttt gccatcgaaa aaccagccca 4440

gttattgggc ttgattggag ctcgctcatt ccaattcctt ctattaggct actaacacca 4500

tgactttatt agcctgtcta tcctggcccc cctggcgagg ttcatgtttg tttatttccg 4560

aatgcaacaa gctccgcatt acacccgaac atcactccag atgagggctt tctgagtgtg 4620

gggtcaaata gtttcatgtt ccccaaatgg cccaaaactg acagtttaaa cgctgtcttg 4680

gaacctaata tgacaaaagc gtgatctcat ccaagatgaa ctaagtttgg ttcgttgaaa 4740

tgctaacggc cagttggtca aaaagaaact tccaaaagtc ggcataccgt ttgtcttgtt 4800

tggtattgat tgacgaatgc tcaaaaataa tctcattaat gcttagcgca gtctctctat 4860

cgcttctgaa ccccggtgca cctgtgccga aacgcaaatg gggaaacacc cgctttttgg 4920

atgattatgc attgtctcca cattgtatgc ttccaagatt ctggtgggaa tactgctgat 4980

agcctaacgt tcatgatcaa aatttaactg ttctaacccc tacttgacag caatatataa 5040

acagaaggaa gctgccctgt cttaaacctt tttttttatc atcattatta gcttactttc 5100

ataattgcga ctggttccaa ttgacaagct tttgatttta acgactttta acgacaactt 5160

gagaagatca aaaaacaact aattattcga aacgatgaga tttccttcaa tttttactgc 5220

tgttttattc gcagcatcct ccgcattagc tgctccagtc aacactacaa cagaagatga 5280

aacggcacaa attccggctg aagctgtcat cggttactca gatttagaag gggatttcga 5340

tgttgctgtt ttgccatttt ccaacagcac aaataacggg ttattgttta taaatactac 5400

tattgccagc attgctgcta aagaagaagg ggtatctctc gagaaaagag aggctgaagc 5460

tgaattccat gaagaggatg gtgtttgtaa ctctaatgct ccatgttacc actgtgatgc 5520

taacggtgaa aactgttctt gtaactgtga attgtttgat tgtgaggcta agaaaccaga 5580

tggttcttat gctcatcctt gtagaagatg tgatgctaac aacatctgta agtgttcttg 5640

tactgctatc ccatgtaatg aagatcatcc ttgtcatcac tgtcacgaag aggatgatgg 5700

agatactcat tgtcactgtt cttgtgagca ttctcacgat catcacgatg atgatactca 5760

tggtgaatgt actaagaaag ctccatgttg gcgttgtgag tacaacgctg atttgaagca 5820

tgatgtttgt ggttgtgaat gttctaaatt gccatgtaat gatgaacacc cttgttatag 5880

aaaggagggt ggtgttgttt cttgtgattg taagactatc acttgtaacg aggatcatcc 5940

ttgttaccac tcttatgaag aggatggtgt tactaagtct gattgtgatt gtgaacattc 6000

tccaggtcct tctgagcatc accatcacca tcactaagcg gccgcgtttg tagccttaga 6060

catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg ctagattcta 6120

atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt ttgatacttt 6180

tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt ctcgtacgag 6240

cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa 6300

atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt acagaagatt 6360

aagtgagacc ttcgtttgtg cggatctaac atccaaagac gaaaggttga atgaaacctt 6420

tttgccatcc gacatccaca ggtccattct cacacataag tgccaaacgc aacaggaggg 6480

gatacactag cagcagaccg ttgcaaacgc aggacctcca ctcctcttct cctcaacacc 6540

cacttttgcc atcgaaaaac cagcccagtt attgggcttg attggagctc gctcattcca 6600

attccttcta ttaggctact aacaccatga ctttattagc ctgtctatcc tggcccccct 6660

ggcgaggttc atgtttgttt atttccgaat gcaacaagct ccgcattaca cccgaacatc 6720

actccagatg agggctttct gagtgtgggg tcaaatagtt tcatgttccc caaatggccc 6780

aaaactgaca gtttaaacgc tgtcttggaa cctaatatga caaaagcgtg atctcatcca 6840

agatgaacta agtttggttc gttgaaatgc taacggccag ttggtcaaaa agaaacttcc 6900

aaaagtcggc ataccgtttg tcttgtttgg tattgattga cgaatgctca aaaataatct 6960

cattaatgct tagcgcagtc tctctatcgc ttctgaaccc cggtgcacct gtgccgaaac 7020

gcaaatgggg aaacacccgc tttttggatg attatgcatt gtctccacat tgtatgcttc 7080

caagattctg gtgggaatac tgctgatagc ctaacgttca tgatcaaaat ttaactgttc 7140

taacccctac ttgacagcaa tatataaaca gaaggaagct gccctgtctt aaaccttttt 7200

ttttatcatc attattagct tactttcata attgcgactg gttccaattg acaagctttt 7260

gattttaacg acttttaacg acaacttgag aagatcaaaa aacaactaat tattcgaaac 7320

gatgagattt ccttcaattt ttactgctgt tttattcgca gcatcctccg cattagctgc 7380

tccagtcaac actacaacag aagatgaaac ggcacaaatt ccggctgaag ctgtcatcgg 7440

ttactcagat ttagaagggg atttcgatgt tgctgttttg ccattttcca acagcacaaa 7500

taacgggtta ttgtttataa atactactat tgccagcatt gctgctaaag aagaaggggt 7560

atctctcgag aaaagagagg ctgaagctga attccatgaa gaggatggtg tttgtaactc 7620

taatgctcca tgttaccact gtgatgctaa cggtgaaaac tgttcttgta actgtgaatt 7680

gtttgattgt gaggctaaga aaccagatgg ttcttatgct catccttgta gaagatgtga 7740

tgctaacaac atctgtaagt gttcttgtac tgctatccca tgtaatgaag atcatccttg 7800

tcatcactgt cacgaagagg atgatggaga tactcattgt cactgttctt gtgagcattc 7860

tcacgatcat cacgatgatg atactcatgg tgaatgtact aagaaagctc catgttggcg 7920

ttgtgagtac aacgctgatt tgaagcatga tgtttgtggt tgtgaatgtt ctaaattgcc 7980

atgtaatgat gaacaccctt gttatagaaa ggagggtggt gttgtttctt gtgattgtaa 8040

gactatcact tgtaacgagg atcatccttg ttaccactct tatgaagagg atggtgttac 8100

taagtctgat tgtgattgtg aacattctcc aggtccttct gagcatcacc atcaccatca 8160

ctaagcggcc gcgtttgtag ccttagacat gactgttcct cagttcaagt tgggcactta 8220

cgagaagacc ggtcttgcta gattctaatc aagaggatgt cagaatgcca tttgcctgag 8280

agatgcaggc ttcatttttg atactttttt atttgtaacc tatatagtat aggatttttt 8340

ttgtcatttt gtttcttctc gtacgagctt gctcctgatc agcctatctc gcagctgatg 8400

aatatcttgt ggtaggggtt tgggaaaatc attcgagttt gatgtttttc ttggtatttc 8460

ccactcctct tcagagtaca gaagattaag tgagaccttc gtttgtgcgg atcctaatgc 8520

ggtagtttat cacagttaaa ttgctaacgc agtcaggcac cgtgtatgaa atctaacaat 8580

gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg 8640

ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat 8700

ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca 8760

ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc 8820

gactacgcga tcatggcgac cacacccgtc ctgtggatct atcgaatcta aatgtaagtt 8880

aaaatctcta aataattaaa taagtcccag tttctccata cgaaccttaa cagcattgcg 8940

gtgagcatct agaccttcaa cagcagccag atccatcact gcttggccaa tatgtttcag 9000

tccctcagga gttacgtctt gtgaagtgat gaacttctgg aaggttgcag tgttaactcc 9060

gctgtattga cgggcatatc cgtacgttgg caaagtgtgg ttggtaccgg aggagtaatc 9120

tccacaactc tctggagagt aggcaccaac aaacacagat ccagcgtgtt gtacttgatc 9180

aacataagaa gaagcattct cgatttgcag gatcaagtgt tcaggagcgt actgattgga 9240

catttccaaa gcctgctcgt aggttgcaac cgatagggtt gtagagtgtg caatacactt 9300

gcgtacaatt tcaacccttg gcaactgcac agcttggttg tgaacagcat cttcaattct 9360

ggcaagctcc ttgtctgtca tatcgacagc caacagaatc acctgggaat caataccatg 9420

ttcagcttga gacagaaggt ctgaggcaac gaaatctgga tcagcgtatt tatcagcaat 9480

aactagaact tcagaaggcc cagcaggcat gtcaatacta cacagggctg atgtgtcatt 9540

ttgaaccatc atcttggcag cagtaacgaa ctggtttcct ggaccaaata ttttgtcaca 9600

cttaggaaca gtttctgttc cgtaagccat agcagctact gcctgggcgc ctcctgctag 9660

cacgatacac ttagcaccaa ccttgtgggc aacgtagatg acttctgggg taagggtacc 9720

atccttctta ggtggagatg caaaaacaat ttctttgcaa ccagcaactt tggcaggaac 9780

acccagcatc agggaagtgg aaggcagaat tgcggttcca ccaggaatat agaggccaac 9840

tttctcaata ggtcttgcaa aacgagagca gactacacca gggcaagtct caacttgcaa 9900

cgtctccgtt agttgagctt catggaattt cctgacgtta tctatagaga gatcaatggc 9960

tctcttaacg ttatctggca attgcataag ttcctctggg aaaggagctt ctaacacagg 10020

tgtcttcaaa gcgactccat caaacttggc agttagttct aaaagggctt tgtcaccatt 10080

ttgacgaaca ttgtcgacaa ttggtttgac taattccata atctgttccg ttttctggat 10140

aggacgacga agggcatctt caatttcttg tgaggaggcc ttagaaacgt caattttgca 10200

caattcaata cgaccttcag aagggacttc tttaggtttg gattcttctt taggttgttc 10260

cttggtgtat cctggcttgg catctccttt ccttctagtg acctttaggg acttcatatc 10320

caggtttctc tccacctcgt ccaacgtcac accgtacttg gcacatctaa ctaatgcaaa 10380

ataaaataag tcagcacatt cccaggctat atcttccttg gatttagctt ctgcaagttc 10440

atcagcttcc tccctaattt tagcgttcaa caaaacttcg tcgtcaaata accgtttggt 10500

ataagaacct tctggagcat tgctcttacg atcccacaag gtggcttcca tggctctaag 10560

accctttgat tggccaaaac aggaagtgcg ttccaagtga cagaaaccaa cacctgtttg 10620

ttcaaccaca aatttcaagc agtctccatc acaatccaat tcgataccca gcaacttttg 10680

agttgctcca gatgtagcac ctttatacca caaaccgtga cgacgagatt ggtagactcc 10740

agtttgtgtc cttatagcct ccggaataga ctttttggac gagtacacca ggcccaacga 10800

gtaattagaa gagtcagcca ccaaagtagt gaatagacca tcggggcggt cagtagtcaa 10860

agacgccaac aaaatttcac tgacagggaa ctttttgaca tcttcagaaa gttcgtattc 10920

agtagtcaat tgccgagcat caataatggg gattatacca gaagcaacag tggaagtcac 10980

atctaccaac tttgcggtct cagaaaaagc ataaacagtt ctactaccgc cattagtgaa 11040

acttttcaaa tcgcccagtg gagaagaaaa aggcacagcg atactagcat tagcgggcaa 11100

ggatgcaact ttatcaacca gggtcctata gataacccta gcgcctggga tcatcctttg 11160

gacaactctt tctgccaaat ctaggtccaa aatcacttca ttgataccat tattgtacaa 11220

cttgagcaag ttgtcgatca gctcctcaaa ttggtcctct gtaacggatg actcaacttg 11280

cacattaact tgaagctcag tcgattgagt gaacttgatc aggttgtgca gctggtcagc 11340

agcataggga aacacggctt ttcctaccaa actcaaggaa ttatcaaact ctgcaacact 11400

tgcgtatgca ggtagcaagg gaaatgtcat acttgaagtc ggacagtgag tgtagtcttg 11460

agaaattctg aagccgtatt tttattatca gtgagtcagt catcaggaga tcctctacgc 11520

cggacgcatc gtggccggca tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc 11580

cgacatcacc gatggggaag atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg 11640

cgtgggtatg gtggcaggcc ccgtggccgg gggactgttg ggcgccatct ccttgcatgc 11700

accattcctt gcggcggcgg tgctcaacgg cctcaaccta ctactgggct gcttcctaat 11760

gcaggagtcg cataagggag agcgtcgagt atctatgatt ggaagtatgg gaatggtgat 11820

acccgcattc ttcagtgtct tgaggtctcc tatcagatta tgcccaacta aagcaaccgg 11880

aggaggagat ttcatggtaa atttctctga cttttggtca tcagtagact cgaactgtga 11940

gactatctcg gttatgacag cagaaatgtc cttcttggag acagtaaatg aagtcccacc 12000

aataaagaaa tccttgttat caggaacaaa cttcttgttt cgaacttttt cggtgccttg 12060

aactataaaa tgtagagtgg atatgtcggg taggaatgga gcgggcaaat gcttaccttc 12120

tggaccttca agaggtatgt agggtttgta gatactgatg ccaacttcag tgacaacgtt 12180

gctatttcgt tcaaaccatt ccgaatccag agaaatcaaa gttgtttgtc tactattgat 12240

ccaagccagt gcggtcttga aactgacaat agtgtgctcg tgttttgagg tcatctttgt 12300

atgaataaat ctagtctttg atctaaataa tcttgacgag ccaaggcgat aaatacccaa 12360

atctaaaact cttttaaaac gttaaaagga caagtatgtc tgcctgtatt aaaccccaaa 12420

tcagctcgta gtctgatcct catcaacttg aggggcacta tcttgtttta gagaaatttg 12480

cggagatgcg atatcgagaa aaaggtacgc tgattttaaa cgtgaaattt atctcaagat 12540

ctgctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga 12600

gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc 12660

agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag tcacgtagcg atagcggagt 12720

gtatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg 12780

tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc 12840

tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca 12900

aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca 12960

aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg 13020

ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg 13080

acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt 13140

ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt 13200

tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc 13260

tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt 13320

gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt 13380

agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc 13440

tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa 13500

agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt 13560

tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct 13620

acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta 13680

tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa 13740

agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc 13800

tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact 13860

acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc 13920

tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt 13980

ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta 14040

agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctgcagg catcgtggtg 14100

tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt 14160

acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc 14220

agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt 14280

actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc 14340

tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaacacg ggataatacc 14400

gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa 14460

ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac 14520

tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa 14580

aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt 14640

tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa 14700

tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct 14760

gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg tatcacgagg 14820

ccctttcgtc ttcaagaatt aattctcatg tttgacagct tatcatcgat aagctgactc 14880

atgttggtat tgtgaaatag acgcagatcg ggaacactga aaaataacag ttattattcg 14940

<210> 3

<211> 19392

<212> DNA

<213> Unknown (Artificial Synthesis)

<400> 3

agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 60

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 120

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 180

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 240

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 300

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 360

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 420

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 480

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 540

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 600

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 660

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 720

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 780

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 840

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 900

caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 960

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 1020

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 1080

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 1140

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 1200

tgaagctgaa ttcgtccccg tgtccggaaa gagcggcagc tcgactaccg ccgtgtcagc 1260

cagcgataat tcggcgctcc cgccactgat atcgtctcgg tgcgcgcctc cgtccaacaa 1320

aggttccaag tctgatttac aggctgagcc ctactacatg cagaaaaaca ccgaatggta 1380

cgagagccat ggcggcaacc taacaagcat aggtaagagg gatgacaatt tggtgggggg 1440

tatgacgctt gacttgccgt cggatgctcc tcctatatcg ctcagtggct ctaccaatag 1500

tgcctcggat ggcggaaaag tagttgcggc gacgacggcg cagatccaag aatttactaa 1560

gtatgcaggg atagccgcaa ctgcatactg ccgcagtgtc gtgcctggca acaaatggga 1620

ctgcgttcag tgccagaagt gggtacccga cggcaagatt attactactt tcacatcact 1680

gttgtctgac acaaatggct atgttttacg atcagacaag caaaaaacaa tctatttagt 1740

cttcagagga actaatagct ttcgttcagc tataacggat attgttttca actttagtga 1800

ctacaaaccc gttaaaggag ccaaggtcca tgctgggttt ctatcgtctt acgagcaagt 1860

tgtaaacgat tacttccctg tcgtgcaaga gcagctgact gcaaacccga catataaagt 1920

aattgtcacg ggtcactctc tgggaggggc ccaggcatta ctggctggga tggatttata 1980

tcaaagggaa ccacggcttt ctccaaagaa cttaagcatt tttaccgtag gcggcccacg 2040

tgtggggaac cccacttttg cttattatgt cgagtctaca gggattccgt ttcagagaac 2100

tgtacacaag cgagatatag tacctcatgt accaccccaa tccttcggat tccttcatcc 2160

aggggtggag tcttggatca aaagtggtac tagtaatgtc caaatttgta cgtccgaaat 2220

agaaaccaaa gactgctcta attctatagt tccctttacg agtatccttg atcacctatc 2280

atatttcgac atcaatgagg gatcttgtct aggcgggggt gggtctggag gtggtggaag 2340

tggcggtggt ggcagtgttc cgcctccatg tgacctgggt atcgccagca aggttaaaca 2400

aaaaggcgta acagggggag gtgcatccgt gtcaacgacc tctgcgactc aaggttctgg 2460

tacaactaat tgtgtcacgc gcaccccgaa ctcggtagag aaaaagaatg tcgcgggcaa 2520

tacgggggtt acagctacca gtgtgtcagc gggcgatgga gctttcggaa atttggctgc 2580

tgcactcaca ctagtcgaag atacagagga cggtctcggt gtaaagacca aaaatggggg 2640

aaagggtttt tccgaaggga ctgccgcgat tagccaaacg gcaggagcaa acgggggggc 2700

cacggtaaaa aaggcgaaac ttgacttgtt gactgatgga gaagaccttt tcgacaccaa 2760

gaaagttgag aagggaaccg tgacctccag ttcctcgcac caaggctcag gtgcagggga 2820

cagcattttt gagatcctca acgaagccga atcgaaaatc aagaagtctg gtgatcacca 2880

ccaccatcat cattgagcgg ccgcgtttgt agccttagac atgactgttc ctcagttcaa 2940

gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc 3000

catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt 3060

ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc 3120

tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt 3180

tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc 3240

ggatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 3300

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 3360

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 3420

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 3480

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 3540

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 3600

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 3660

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 3720

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 3780

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 3840

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 3900

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 3960

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 4020

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 4080

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 4140

caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 4200

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 4260

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 4320

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 4380

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 4440

tgaagctgaa ttcgtccccg tgtccggaaa gagcggcagc tcgactaccg ccgtgtcagc 4500

cagcgataat tcggcgctcc cgccactgat atcgtctcgg tgcgcgcctc cgtccaacaa 4560

aggttccaag tctgatttac aggctgagcc ctactacatg cagaaaaaca ccgaatggta 4620

cgagagccat ggcggcaacc taacaagcat aggtaagagg gatgacaatt tggtgggggg 4680

tatgacgctt gacttgccgt cggatgctcc tcctatatcg ctcagtggct ctaccaatag 4740

tgcctcggat ggcggaaaag tagttgcggc gacgacggcg cagatccaag aatttactaa 4800

gtatgcaggg atagccgcaa ctgcatactg ccgcagtgtc gtgcctggca acaaatggga 4860

ctgcgttcag tgccagaagt gggtacccga cggcaagatt attactactt tcacatcact 4920

gttgtctgac acaaatggct atgttttacg atcagacaag caaaaaacaa tctatttagt 4980

cttcagagga actaatagct ttcgttcagc tataacggat attgttttca actttagtga 5040

ctacaaaccc gttaaaggag ccaaggtcca tgctgggttt ctatcgtctt acgagcaagt 5100

tgtaaacgat tacttccctg tcgtgcaaga gcagctgact gcaaacccga catataaagt 5160

aattgtcacg ggtcactctc tgggaggggc ccaggcatta ctggctggga tggatttata 5220

tcaaagggaa ccacggcttt ctccaaagaa cttaagcatt tttaccgtag gcggcccacg 5280

tgtggggaac cccacttttg cttattatgt cgagtctaca gggattccgt ttcagagaac 5340

tgtacacaag cgagatatag tacctcatgt accaccccaa tccttcggat tccttcatcc 5400

aggggtggag tcttggatca aaagtggtac tagtaatgtc caaatttgta cgtccgaaat 5460

agaaaccaaa gactgctcta attctatagt tccctttacg agtatccttg atcacctatc 5520

atatttcgac atcaatgagg gatcttgtct aggcgggggt gggtctggag gtggtggaag 5580

tggcggtggt ggcagtgttc cgcctccatg tgacctgggt atcgccagca aggttaaaca 5640

aaaaggcgta acagggggag gtgcatccgt gtcaacgacc tctgcgactc aaggttctgg 5700

tacaactaat tgtgtcacgc gcaccccgaa ctcggtagag aaaaagaatg tcgcgggcaa 5760

tacgggggtt acagctacca gtgtgtcagc gggcgatgga gctttcggaa atttggctgc 5820

tgcactcaca ctagtcgaag atacagagga cggtctcggt gtaaagacca aaaatggggg 5880

aaagggtttt tccgaaggga ctgccgcgat tagccaaacg gcaggagcaa acgggggggc 5940

cacggtaaaa aaggcgaaac ttgacttgtt gactgatgga gaagaccttt tcgacaccaa 6000

gaaagttgag aagggaaccg tgacctccag ttcctcgcac caaggctcag gtgcagggga 6060

cagcattttt gagatcctca acgaagccga atcgaaaatc aagaagtctg gtgatcacca 6120

ccaccatcat cattgagcgg ccgcgtttgt agccttagac atgactgttc ctcagttcaa 6180

gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc 6240

catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt 6300

ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc 6360

tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt 6420

tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc 6480

ggatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 6540

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 6600

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 6660

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 6720

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 6780

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 6840

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 6900

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 6960

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 7020

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 7080

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 7140

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 7200

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 7260

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 7320

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 7380

caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 7440

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 7500

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 7560

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 7620

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 7680

tgaagctgaa ttcgtccccg tgtccggaaa gagcggcagc tcgactaccg ccgtgtcagc 7740

cagcgataat tcggcgctcc cgccactgat atcgtctcgg tgcgcgcctc cgtccaacaa 7800

aggttccaag tctgatttac aggctgagcc ctactacatg cagaaaaaca ccgaatggta 7860

cgagagccat ggcggcaacc taacaagcat aggtaagagg gatgacaatt tggtgggggg 7920

tatgacgctt gacttgccgt cggatgctcc tcctatatcg ctcagtggct ctaccaatag 7980

tgcctcggat ggcggaaaag tagttgcggc gacgacggcg cagatccaag aatttactaa 8040

gtatgcaggg atagccgcaa ctgcatactg ccgcagtgtc gtgcctggca acaaatggga 8100

ctgcgttcag tgccagaagt gggtacccga cggcaagatt attactactt tcacatcact 8160

gttgtctgac acaaatggct atgttttacg atcagacaag caaaaaacaa tctatttagt 8220

cttcagagga actaatagct ttcgttcagc tataacggat attgttttca actttagtga 8280

ctacaaaccc gttaaaggag ccaaggtcca tgctgggttt ctatcgtctt acgagcaagt 8340

tgtaaacgat tacttccctg tcgtgcaaga gcagctgact gcaaacccga catataaagt 8400

aattgtcacg ggtcactctc tgggaggggc ccaggcatta ctggctggga tggatttata 8460

tcaaagggaa ccacggcttt ctccaaagaa cttaagcatt tttaccgtag gcggcccacg 8520

tgtggggaac cccacttttg cttattatgt cgagtctaca gggattccgt ttcagagaac 8580

tgtacacaag cgagatatag tacctcatgt accaccccaa tccttcggat tccttcatcc 8640

aggggtggag tcttggatca aaagtggtac tagtaatgtc caaatttgta cgtccgaaat 8700

agaaaccaaa gactgctcta attctatagt tccctttacg agtatccttg atcacctatc 8760

atatttcgac atcaatgagg gatcttgtct aggcgggggt gggtctggag gtggtggaag 8820

tggcggtggt ggcagtgttc cgcctccatg tgacctgggt atcgccagca aggttaaaca 8880

aaaaggcgta acagggggag gtgcatccgt gtcaacgacc tctgcgactc aaggttctgg 8940

tacaactaat tgtgtcacgc gcaccccgaa ctcggtagag aaaaagaatg tcgcgggcaa 9000

tacgggggtt acagctacca gtgtgtcagc gggcgatgga gctttcggaa atttggctgc 9060

tgcactcaca ctagtcgaag atacagagga cggtctcggt gtaaagacca aaaatggggg 9120

aaagggtttt tccgaaggga ctgccgcgat tagccaaacg gcaggagcaa acgggggggc 9180

cacggtaaaa aaggcgaaac ttgacttgtt gactgatgga gaagaccttt tcgacaccaa 9240

gaaagttgag aagggaaccg tgacctccag ttcctcgcac caaggctcag gtgcagggga 9300

cagcattttt gagatcctca acgaagccga atcgaaaatc aagaagtctg gtgatcacca 9360

ccaccatcat cattgagcgg ccgcgtttgt agccttagac atgactgttc ctcagttcaa 9420

gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc 9480

catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt 9540

ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc 9600

tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt 9660

tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc 9720

ggatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 9780

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 9840

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 9900

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 9960

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 10020

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 10080

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 10140

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 10200

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 10260

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 10320

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 10380

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 10440

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 10500

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 10560

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 10620

caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 10680

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 10740

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 10800

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 10860

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 10920

tgaagctgaa ttcgtccccg tgtccggaaa gagcggcagc tcgactaccg ccgtgtcagc 10980

cagcgataat tcggcgctcc cgccactgat atcgtctcgg tgcgcgcctc cgtccaacaa 11040

aggttccaag tctgatttac aggctgagcc ctactacatg cagaaaaaca ccgaatggta 11100

cgagagccat ggcggcaacc taacaagcat aggtaagagg gatgacaatt tggtgggggg 11160

tatgacgctt gacttgccgt cggatgctcc tcctatatcg ctcagtggct ctaccaatag 11220

tgcctcggat ggcggaaaag tagttgcggc gacgacggcg cagatccaag aatttactaa 11280

gtatgcaggg atagccgcaa ctgcatactg ccgcagtgtc gtgcctggca acaaatggga 11340

ctgcgttcag tgccagaagt gggtacccga cggcaagatt attactactt tcacatcact 11400

gttgtctgac acaaatggct atgttttacg atcagacaag caaaaaacaa tctatttagt 11460

cttcagagga actaatagct ttcgttcagc tataacggat attgttttca actttagtga 11520

ctacaaaccc gttaaaggag ccaaggtcca tgctgggttt ctatcgtctt acgagcaagt 11580

tgtaaacgat tacttccctg tcgtgcaaga gcagctgact gcaaacccga catataaagt 11640

aattgtcacg ggtcactctc tgggaggggc ccaggcatta ctggctggga tggatttata 11700

tcaaagggaa ccacggcttt ctccaaagaa cttaagcatt tttaccgtag gcggcccacg 11760

tgtggggaac cccacttttg cttattatgt cgagtctaca gggattccgt ttcagagaac 11820

tgtacacaag cgagatatag tacctcatgt accaccccaa tccttcggat tccttcatcc 11880

aggggtggag tcttggatca aaagtggtac tagtaatgtc caaatttgta cgtccgaaat 11940

agaaaccaaa gactgctcta attctatagt tccctttacg agtatccttg atcacctatc 12000

atatttcgac atcaatgagg gatcttgtct aggcgggggt gggtctggag gtggtggaag 12060

tggcggtggt ggcagtgttc cgcctccatg tgacctgggt atcgccagca aggttaaaca 12120

aaaaggcgta acagggggag gtgcatccgt gtcaacgacc tctgcgactc aaggttctgg 12180

tacaactaat tgtgtcacgc gcaccccgaa ctcggtagag aaaaagaatg tcgcgggcaa 12240

tacgggggtt acagctacca gtgtgtcagc gggcgatgga gctttcggaa atttggctgc 12300

tgcactcaca ctagtcgaag atacagagga cggtctcggt gtaaagacca aaaatggggg 12360

aaagggtttt tccgaaggga ctgccgcgat tagccaaacg gcaggagcaa acgggggggc 12420

cacggtaaaa aaggcgaaac ttgacttgtt gactgatgga gaagaccttt tcgacaccaa 12480

gaaagttgag aagggaaccg tgacctccag ttcctcgcac caaggctcag gtgcagggga 12540

cagcattttt gagatcctca acgaagccga atcgaaaatc aagaagtctg gtgatcacca 12600

ccaccatcat cattgagcgg ccgcgtttgt agccttagac atgactgttc ctcagttcaa 12660

gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc 12720

catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt 12780

ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc 12840

tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt 12900

tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc 12960

ggatcctaat gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg 13020

aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata 13080

ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc 13140

gccagtcact atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc 13200

gttctcggag cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt 13260

ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat ctatcgaatc 13320

taaatgtaag ttaaaatctc taaataatta aataagtccc agtttctcca tacgaacctt 13380

aacagcattg cggtgagcat ctagaccttc aacagcagcc agatccatca ctgcttggcc 13440

aatatgtttc agtccctcag gagttacgtc ttgtgaagtg atgaacttct ggaaggttgc 13500

agtgttaact ccgctgtatt gacgggcata tccgtacgtt ggcaaagtgt ggttggtacc 13560

ggaggagtaa tctccacaac tctctggaga gtaggcacca acaaacacag atccagcgtg 13620

ttgtacttga tcaacataag aagaagcatt ctcgatttgc aggatcaagt gttcaggagc 13680

gtactgattg gacatttcca aagcctgctc gtaggttgca accgataggg ttgtagagtg 13740

tgcaatacac ttgcgtacaa tttcaaccct tggcaactgc acagcttggt tgtgaacagc 13800

atcttcaatt ctggcaagct ccttgtctgt catatcgaca gccaacagaa tcacctggga 13860

atcaatacca tgttcagctt gagacagaag gtctgaggca acgaaatctg gatcagcgta 13920

tttatcagca ataactagaa cttcagaagg cccagcaggc atgtcaatac tacacagggc 13980

tgatgtgtca ttttgaacca tcatcttggc agcagtaacg aactggtttc ctggaccaaa 14040

tattttgtca cacttaggaa cagtttctgt tccgtaagcc atagcagcta ctgcctgggc 14100

gcctcctgct agcacgatac acttagcacc aaccttgtgg gcaacgtaga tgacttctgg 14160

ggtaagggta ccatccttct taggtggaga tgcaaaaaca atttctttgc aaccagcaac 14220

tttggcagga acacccagca tcagggaagt ggaaggcaga attgcggttc caccaggaat 14280

atagaggcca actttctcaa taggtcttgc aaaacgagag cagactacac cagggcaagt 14340

ctcaacttgc aacgtctccg ttagttgagc ttcatggaat ttcctgacgt tatctataga 14400

gagatcaatg gctctcttaa cgttatctgg caattgcata agttcctctg ggaaaggagc 14460

ttctaacaca ggtgtcttca aagcgactcc atcaaacttg gcagttagtt ctaaaagggc 14520

tttgtcacca ttttgacgaa cattgtcgac aattggtttg actaattcca taatctgttc 14580

cgttttctgg ataggacgac gaagggcatc ttcaatttct tgtgaggagg ccttagaaac 14640

gtcaattttg cacaattcaa tacgaccttc agaagggact tctttaggtt tggattcttc 14700

tttaggttgt tccttggtgt atcctggctt ggcatctcct ttccttctag tgacctttag 14760

ggacttcata tccaggtttc tctccacctc gtccaacgtc acaccgtact tggcacatct 14820

aactaatgca aaataaaata agtcagcaca ttcccaggct atatcttcct tggatttagc 14880

ttctgcaagt tcatcagctt cctccctaat tttagcgttc aacaaaactt cgtcgtcaaa 14940

taaccgtttg gtataagaac cttctggagc attgctctta cgatcccaca aggtggcttc 15000

catggctcta agaccctttg attggccaaa acaggaagtg cgttccaagt gacagaaacc 15060

aacacctgtt tgttcaacca caaatttcaa gcagtctcca tcacaatcca attcgatacc 15120

cagcaacttt tgagttgctc cagatgtagc acctttatac cacaaaccgt gacgacgaga 15180

ttggtagact ccagtttgtg tccttatagc ctccggaata gactttttgg acgagtacac 15240

caggcccaac gagtaattag aagagtcagc caccaaagta gtgaatagac catcggggcg 15300

gtcagtagtc aaagacgcca acaaaatttc actgacaggg aactttttga catcttcaga 15360

aagttcgtat tcagtagtca attgccgagc atcaataatg gggattatac cagaagcaac 15420

agtggaagtc acatctacca actttgcggt ctcagaaaaa gcataaacag ttctactacc 15480

gccattagtg aaacttttca aatcgcccag tggagaagaa aaaggcacag cgatactagc 15540

attagcgggc aaggatgcaa ctttatcaac cagggtccta tagataaccc tagcgcctgg 15600

gatcatcctt tggacaactc tttctgccaa atctaggtcc aaaatcactt cattgatacc 15660

attattgtac aacttgagca agttgtcgat cagctcctca aattggtcct ctgtaacgga 15720

tgactcaact tgcacattaa cttgaagctc agtcgattga gtgaacttga tcaggttgtg 15780

cagctggtca gcagcatagg gaaacacggc ttttcctacc aaactcaagg aattatcaaa 15840

ctctgcaaca cttgcgtatg caggtagcaa gggaaatgtc atacttgaag tcggacagtg 15900

agtgtagtct tgagaaattc tgaagccgta tttttattat cagtgagtca gtcatcagga 15960

gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg 16020

cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag 16080

cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat 16140

ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg 16200

ctgcttccta atgcaggagt cgcataaggg agagcgtcga gtatctatga ttggaagtat 16260

gggaatggtg atacccgcat tcttcagtgt cttgaggtct cctatcagat tatgcccaac 16320

taaagcaacc ggaggaggag atttcatggt aaatttctct gacttttggt catcagtaga 16380

ctcgaactgt gagactatct cggttatgac agcagaaatg tccttcttgg agacagtaaa 16440

tgaagtccca ccaataaaga aatccttgtt atcaggaaca aacttcttgt ttcgaacttt 16500

ttcggtgcct tgaactataa aatgtagagt ggatatgtcg ggtaggaatg gagcgggcaa 16560

atgcttacct tctggacctt caagaggtat gtagggtttg tagatactga tgccaacttc 16620

agtgacaacg ttgctatttc gttcaaacca ttccgaatcc agagaaatca aagttgtttg 16680

tctactattg atccaagcca gtgcggtctt gaaactgaca atagtgtgct cgtgttttga 16740

ggtcatcttt gtatgaataa atctagtctt tgatctaaat aatcttgacg agccaaggcg 16800

ataaataccc aaatctaaaa ctcttttaaa acgttaaaag gacaagtatg tctgcctgta 16860

ttaaacccca aatcagctcg tagtctgatc ctcatcaact tgaggggcac tatcttgttt 16920

tagagaaatt tgcggagatg cgatatcgag aaaaaggtac gctgatttta aacgtgaaat 16980

ttatctcaag atctgctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 17040

gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 17100

tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag 17160

cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg 17220

caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc 17280

tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 17340

tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 17400

aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 17460

tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 17520

tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 17580

cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 17640

agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 17700

tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 17760

aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 17820

ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 17880

cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 17940

accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 18000

ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 18060

ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 18120

gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 18180

aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 18240

gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 18300

gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 18360

cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 18420

gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 18480

gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca 18540

ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 18600

tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 18660

ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 18720

cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 18780

accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca 18840

cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 18900

tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 18960

cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 19020

acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 19080

atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 19140

tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 19200

aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 19260

cgtatcacga ggccctttcg tcttcaagaa ttaattctca tgtttgacag cttatcatcg 19320

ataagctgac tcatgttggt attgtgaaat agacgcagat cgggaacact gaaaaataac 19380

agttattatt cg 19392

<210> 4

<211> 19512

<212> DNA

<213> unknown (Artificial Synthesis)

<400> 4

agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 60

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 120

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 180

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 240

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 300

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 360

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 420

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 480

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 540

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 600

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 660

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 720

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 780

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 840

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 900

caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 960

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 1020

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 1080

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 1140

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 1200

tgaagctgaa ttcgtcccag tctccggtaa gtctgggtca agtactacag cagtcagtgc 1260

ttcggacaac tccgcgcttc cgccgctgat tagttcgcga tgtgctccgc cctctaacaa 1320

gggtagtaaa tctgacttac aggcggagcc atattacatg cagaaaaaca ccgagtggta 1380

tgagagccac ggcggtaact tgaccagtat cgggaagcgc gatgacaact tagttggggg 1440

gatgacactg gatctgccta gcgatgcacc ccccatatct ttgtcgggat cgactaattc 1500

tgcaagcgac ggtggaaagg tagttgcagc tacgacggct caaatccagg agttcacaaa 1560

atacgccgga atagcggcca cggcttattg tcggtctgta gtcccaggga ataagtggga 1620

ctgcgtccag tgccagaaat gggtacctga cggtaagatt atcactacat ttaccagttt 1680

actgtcggat acgaatgggt acgttctaag gagcgacaaa caaaaaacca tctatctagt 1740

ttttagaggc acaaactcgt tcagaagcgc cattaccgac atcgtattta attttagcga 1800

ctataagccc gttaagggtg cgaaagtaca tgccggcttt cttagttcat atgaacaggt 1860

ggtgaatgac tacttcccgg tggtgcaaga acagctcact gcgaatccta catacaaagt 1920

catcgttact ggacacagct tagggggtgc acaagccctt ctcgccggaa tggacctata 1980

tcaacgggag cctcgactct ccccaaagaa tcttagtata ttcactgtag gcggcccccg 2040

cgtaggcaac cccacctttg cttattacgt ggaatctact ggtattcctt tccaacggac 2100

ggttcataag cgcgacatag taccacacgt tccgccccag tccttcgggt tcttgcatcc 2160

aggcgtggag tcatggatta aatcgggtac gtcaaatgtc caaatctgta catcagaaat 2220

tgaaacaaaa gattgttcaa actcaatagt tccgtttaca tcgattctgg atcacttgtc 2280

atactttgac attaacgaag gttcgtgctt gggcggcgga ggatctggtg gcggcgggtc 2340

cggcggaggt gggagccatg aagaggacgg tgtctgcaac tcaaatgcgc catgttatca 2400

ctgtgatgca aacggggaaa attgctcctg taactgcgag ctattcgatt gcgaagcgaa 2460

aaaacccgac ggttcttatg cccacccatg ccgtcgatgt gatgcgaata atatatgtaa 2520

gtgttcctgc acggctatac cttgtaatga ggaccatcca tgtcaccact gtcatgagga 2580

agatgatggt gacacccact gtcactgtag ttgtgagcac tcccacgatc atcacgatga 2640

cgatactcat ggggagtgca ctaagaaggc accgtgttgg aggtgtgaat acaatgccga 2700

tctcaagcat gatgtctgcg gatgcgagtg cagtaaacta ccttgtaacg acgagcatcc 2760

ctgctatcgt aaagaaggag gagtggtgtc ttgcgactgc aagacgataa cctgcaacga 2820

agatcatccg tgttatcaca gctacgaaga agatggagta accaaatcgg attgcgattg 2880

cgagcactcc cctggtcctt ctgaacatca ccatcatcat cattgagcgg ccgcgtttgt 2940

agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc 3000

tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt 3060

tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc 3120

tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg 3180

tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta 3240

cagaagatta agtgagacct tcgtttgtgc ggatctaaca tccaaagacg aaaggttgaa 3300

tgaaaccttt ttgccatccg acatccacag gtccattctc acacataagt gccaaacgca 3360

acaggagggg atacactagc agcagaccgt tgcaaacgca ggacctccac tcctcttctc 3420

ctcaacaccc acttttgcca tcgaaaaacc agcccagtta ttgggcttga ttggagctcg 3480

ctcattccaa ttccttctat taggctacta acaccatgac tttattagcc tgtctatcct 3540

ggcccccctg gcgaggttca tgtttgttta tttccgaatg caacaagctc cgcattacac 3600

ccgaacatca ctccagatga gggctttctg agtgtggggt caaatagttt catgttcccc 3660

aaatggccca aaactgacag tttaaacgct gtcttggaac ctaatatgac aaaagcgtga 3720

tctcatccaa gatgaactaa gtttggttcg ttgaaatgct aacggccagt tggtcaaaaa 3780

gaaacttcca aaagtcggca taccgtttgt cttgtttggt attgattgac gaatgctcaa 3840

aaataatctc attaatgctt agcgcagtct ctctatcgct tctgaacccc ggtgcacctg 3900

tgccgaaacg caaatgggga aacacccgct ttttggatga ttatgcattg tctccacatt 3960

gtatgcttcc aagattctgg tgggaatact gctgatagcc taacgttcat gatcaaaatt 4020

taactgttct aacccctact tgacagcaat atataaacag aaggaagctg ccctgtctta 4080

aacctttttt tttatcatca ttattagctt actttcataa ttgcgactgg ttccaattga 4140

caagcttttg attttaacga cttttaacga caacttgaga agatcaaaaa acaactaatt 4200

attcgaaacg atgagatttc cttcaatttt tactgctgtt ttattcgcag catcctccgc 4260

attagctgct ccagtcaaca ctacaacaga agatgaaacg gcacaaattc cggctgaagc 4320

tgtcatcggt tactcagatt tagaagggga tttcgatgtt gctgttttgc cattttccaa 4380

cagcacaaat aacgggttat tgtttataaa tactactatt gccagcattg ctgctaaaga 4440

agaaggggta tctctcgaga aaagagaggc tgaagctgaa ttcgtcccag tctccggtaa 4500

gtctgggtca agtactacag cagtcagtgc ttcggacaac tccgcgcttc cgccgctgat 4560

tagttcgcga tgtgctccgc cctctaacaa gggtagtaaa tctgacttac aggcggagcc 4620

atattacatg cagaaaaaca ccgagtggta tgagagccac ggcggtaact tgaccagtat 4680

cgggaagcgc gatgacaact tagttggggg gatgacactg gatctgccta gcgatgcacc 4740

ccccatatct ttgtcgggat cgactaattc tgcaagcgac ggtggaaagg tagttgcagc 4800

tacgacggct caaatccagg agttcacaaa atacgccgga atagcggcca cggcttattg 4860

tcggtctgta gtcccaggga ataagtggga ctgcgtccag tgccagaaat gggtacctga 4920

cggtaagatt atcactacat ttaccagttt actgtcggat acgaatgggt acgttctaag 4980

gagcgacaaa caaaaaacca tctatctagt ttttagaggc acaaactcgt tcagaagcgc 5040

cattaccgac atcgtattta attttagcga ctataagccc gttaagggtg cgaaagtaca 5100

tgccggcttt cttagttcat atgaacaggt ggtgaatgac tacttcccgg tggtgcaaga 5160

acagctcact gcgaatccta catacaaagt catcgttact ggacacagct tagggggtgc 5220

acaagccctt ctcgccggaa tggacctata tcaacgggag cctcgactct ccccaaagaa 5280

tcttagtata ttcactgtag gcggcccccg cgtaggcaac cccacctttg cttattacgt 5340

ggaatctact ggtattcctt tccaacggac ggttcataag cgcgacatag taccacacgt 5400

tccgccccag tccttcgggt tcttgcatcc aggcgtggag tcatggatta aatcgggtac 5460

gtcaaatgtc caaatctgta catcagaaat tgaaacaaaa gattgttcaa actcaatagt 5520

tccgtttaca tcgattctgg atcacttgtc atactttgac attaacgaag gttcgtgctt 5580

gggcggcgga ggatctggtg gcggcgggtc cggcggaggt gggagccatg aagaggacgg 5640

tgtctgcaac tcaaatgcgc catgttatca ctgtgatgca aacggggaaa attgctcctg 5700

taactgcgag ctattcgatt gcgaagcgaa aaaacccgac ggttcttatg cccacccatg 5760

ccgtcgatgt gatgcgaata atatatgtaa gtgttcctgc acggctatac cttgtaatga 5820

ggaccatcca tgtcaccact gtcatgagga agatgatggt gacacccact gtcactgtag 5880

ttgtgagcac tcccacgatc atcacgatga cgatactcat ggggagtgca ctaagaaggc 5940

accgtgttgg aggtgtgaat acaatgccga tctcaagcat gatgtctgcg gatgcgagtg 6000

cagtaaacta ccttgtaacg acgagcatcc ctgctatcgt aaagaaggag gagtggtgtc 6060

ttgcgactgc aagacgataa cctgcaacga agatcatccg tgttatcaca gctacgaaga 6120

agatggagta accaaatcgg attgcgattg cgagcactcc cctggtcctt ctgaacatca 6180

ccatcatcat cattgagcgg ccgcgtttgt agccttagac atgactgttc ctcagttcaa 6240

gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc 6300

catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt 6360

ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc 6420

tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt 6480

tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc 6540

ggatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 6600

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 6660

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 6720

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 6780

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 6840

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 6900

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 6960

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 7020

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 7080

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 7140

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 7200

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 7260

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 7320

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 7380

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 7440

caacttgaga agatcaaaaa acaactaatt attcgaaacg atgagatttc cttcaatttt 7500

tactgctgtt ttattcgcag catcctccgc attagctgct ccagtcaaca ctacaacaga 7560

agatgaaacg gcacaaattc cggctgaagc tgtcatcggt tactcagatt tagaagggga 7620

tttcgatgtt gctgttttgc cattttccaa cagcacaaat aacgggttat tgtttataaa 7680

tactactatt gccagcattg ctgctaaaga agaaggggta tctctcgaga aaagagaggc 7740

tgaagctgaa ttcgtcccag tctccggtaa gtctgggtca agtactacag cagtcagtgc 7800

ttcggacaac tccgcgcttc cgccgctgat tagttcgcga tgtgctccgc cctctaacaa 7860

gggtagtaaa tctgacttac aggcggagcc atattacatg cagaaaaaca ccgagtggta 7920

tgagagccac ggcggtaact tgaccagtat cgggaagcgc gatgacaact tagttggggg 7980

gatgacactg gatctgccta gcgatgcacc ccccatatct ttgtcgggat cgactaattc 8040

tgcaagcgac ggtggaaagg tagttgcagc tacgacggct caaatccagg agttcacaaa 8100

atacgccgga atagcggcca cggcttattg tcggtctgta gtcccaggga ataagtggga 8160

ctgcgtccag tgccagaaat gggtacctga cggtaagatt atcactacat ttaccagttt 8220

actgtcggat acgaatgggt acgttctaag gagcgacaaa caaaaaacca tctatctagt 8280

ttttagaggc acaaactcgt tcagaagcgc cattaccgac atcgtattta attttagcga 8340

ctataagccc gttaagggtg cgaaagtaca tgccggcttt cttagttcat atgaacaggt 8400

ggtgaatgac tacttcccgg tggtgcaaga acagctcact gcgaatccta catacaaagt 8460

catcgttact ggacacagct tagggggtgc acaagccctt ctcgccggaa tggacctata 8520

tcaacgggag cctcgactct ccccaaagaa tcttagtata ttcactgtag gcggcccccg 8580

cgtaggcaac cccacctttg cttattacgt ggaatctact ggtattcctt tccaacggac 8640

ggttcataag cgcgacatag taccacacgt tccgccccag tccttcgggt tcttgcatcc 8700

aggcgtggag tcatggatta aatcgggtac gtcaaatgtc caaatctgta catcagaaat 8760

tgaaacaaaa gattgttcaa actcaatagt tccgtttaca tcgattctgg atcacttgtc 8820

atactttgac attaacgaag gttcgtgctt gggcggcgga ggatctggtg gcggcgggtc 8880

cggcggaggt gggagccatg aagaggacgg tgtctgcaac tcaaatgcgc catgttatca 8940

ctgtgatgca aacggggaaa attgctcctg taactgcgag ctattcgatt gcgaagcgaa 9000

aaaacccgac ggttcttatg cccacccatg ccgtcgatgt gatgcgaata atatatgtaa 9060

gtgttcctgc acggctatac cttgtaatga ggaccatcca tgtcaccact gtcatgagga 9120

agatgatggt gacacccact gtcactgtag ttgtgagcac tcccacgatc atcacgatga 9180

cgatactcat ggggagtgca ctaagaaggc accgtgttgg aggtgtgaat acaatgccga 9240

tctcaagcat gatgtctgcg gatgcgagtg cagtaaacta ccttgtaacg acgagcatcc 9300

ctgctatcgt aaagaaggag gagtggtgtc ttgcgactgc aagacgataa cctgcaacga 9360

agatcatccg tgttatcaca gctacgaaga agatggagta accaaatcgg attgcgattg 9420

cgagcactcc cctggtcctt ctgaacatca ccatcatcat cattgagcgg ccgcgtttgt 9480

agccttagac atgactgttc ctcagttcaa gttgggcact tacgagaaga ccggtcttgc 9540

tagattctaa tcaagaggat gtcagaatgc catttgcctg agagatgcag gcttcatttt 9600

tgatactttt ttatttgtaa cctatatagt ataggatttt ttttgtcatt ttgtttcttc 9660

tcgtacgagc ttgctcctga tcagcctatc tcgcagctga tgaatatctt gtggtagggg 9720

tttgggaaaa tcattcgagt ttgatgtttt tcttggtatt tcccactcct cttcagagta 9780

cagaagatta agtgagacct tcgtttgtgc ggatctaaca tccaaagacg aaaggttgaa 9840

tgaaaccttt ttgccatccg acatccacag gtccattctc acacataagt gccaaacgca 9900

acaggagggg atacactagc agcagaccgt tgcaaacgca ggacctccac tcctcttctc 9960

ctcaacaccc acttttgcca tcgaaaaacc agcccagtta ttgggcttga ttggagctcg 10020

ctcattccaa ttccttctat taggctacta acaccatgac tttattagcc tgtctatcct 10080

ggcccccctg gcgaggttca tgtttgttta tttccgaatg caacaagctc cgcattacac 10140

ccgaacatca ctccagatga gggctttctg agtgtggggt caaatagttt catgttcccc 10200

aaatggccca aaactgacag tttaaacgct gtcttggaac ctaatatgac aaaagcgtga 10260

tctcatccaa gatgaactaa gtttggttcg ttgaaatgct aacggccagt tggtcaaaaa 10320

gaaacttcca aaagtcggca taccgtttgt cttgtttggt attgattgac gaatgctcaa 10380

aaataatctc attaatgctt agcgcagtct ctctatcgct tctgaacccc ggtgcacctg 10440

tgccgaaacg caaatgggga aacacccgct ttttggatga ttatgcattg tctccacatt 10500

gtatgcttcc aagattctgg tgggaatact gctgatagcc taacgttcat gatcaaaatt 10560

taactgttct aacccctact tgacagcaat atataaacag aaggaagctg ccctgtctta 10620

aacctttttt tttatcatca ttattagctt actttcataa ttgcgactgg ttccaattga 10680

caagcttttg attttaacga cttttaacga caacttgaga agatcaaaaa acaactaatt 10740

attcgaaacg atgagatttc cttcaatttt tactgctgtt ttattcgcag catcctccgc 10800

attagctgct ccagtcaaca ctacaacaga agatgaaacg gcacaaattc cggctgaagc 10860

tgtcatcggt tactcagatt tagaagggga tttcgatgtt gctgttttgc cattttccaa 10920

cagcacaaat aacgggttat tgtttataaa tactactatt gccagcattg ctgctaaaga 10980

agaaggggta tctctcgaga aaagagaggc tgaagctgaa ttcgtcccag tctccggtaa 11040

gtctgggtca agtactacag cagtcagtgc ttcggacaac tccgcgcttc cgccgctgat 11100

tagttcgcga tgtgctccgc cctctaacaa gggtagtaaa tctgacttac aggcggagcc 11160

atattacatg cagaaaaaca ccgagtggta tgagagccac ggcggtaact tgaccagtat 11220

cgggaagcgc gatgacaact tagttggggg gatgacactg gatctgccta gcgatgcacc 11280

ccccatatct ttgtcgggat cgactaattc tgcaagcgac ggtggaaagg tagttgcagc 11340

tacgacggct caaatccagg agttcacaaa atacgccgga atagcggcca cggcttattg 11400

tcggtctgta gtcccaggga ataagtggga ctgcgtccag tgccagaaat gggtacctga 11460

cggtaagatt atcactacat ttaccagttt actgtcggat acgaatgggt acgttctaag 11520

gagcgacaaa caaaaaacca tctatctagt ttttagaggc acaaactcgt tcagaagcgc 11580

cattaccgac atcgtattta attttagcga ctataagccc gttaagggtg cgaaagtaca 11640

tgccggcttt cttagttcat atgaacaggt ggtgaatgac tacttcccgg tggtgcaaga 11700

acagctcact gcgaatccta catacaaagt catcgttact ggacacagct tagggggtgc 11760

acaagccctt ctcgccggaa tggacctata tcaacgggag cctcgactct ccccaaagaa 11820

tcttagtata ttcactgtag gcggcccccg cgtaggcaac cccacctttg cttattacgt 11880

ggaatctact ggtattcctt tccaacggac ggttcataag cgcgacatag taccacacgt 11940

tccgccccag tccttcgggt tcttgcatcc aggcgtggag tcatggatta aatcgggtac 12000

gtcaaatgtc caaatctgta catcagaaat tgaaacaaaa gattgttcaa actcaatagt 12060

tccgtttaca tcgattctgg atcacttgtc atactttgac attaacgaag gttcgtgctt 12120

gggcggcgga ggatctggtg gcggcgggtc cggcggaggt gggagccatg aagaggacgg 12180

tgtctgcaac tcaaatgcgc catgttatca ctgtgatgca aacggggaaa attgctcctg 12240

taactgcgag ctattcgatt gcgaagcgaa aaaacccgac ggttcttatg cccacccatg 12300

ccgtcgatgt gatgcgaata atatatgtaa gtgttcctgc acggctatac cttgtaatga 12360

ggaccatcca tgtcaccact gtcatgagga agatgatggt gacacccact gtcactgtag 12420

ttgtgagcac tcccacgatc atcacgatga cgatactcat ggggagtgca ctaagaaggc 12480

accgtgttgg aggtgtgaat acaatgccga tctcaagcat gatgtctgcg gatgcgagtg 12540

cagtaaacta ccttgtaacg acgagcatcc ctgctatcgt aaagaaggag gagtggtgtc 12600

ttgcgactgc aagacgataa cctgcaacga agatcatccg tgttatcaca gctacgaaga 12660

agatggagta accaaatcgg attgcgattg cgagcactcc cctggtcctt ctgaacatca 12720

ccatcatcat cattgagcgg ccgcgtttgt agccttagac atgactgttc ctcagttcaa 12780

gttgggcact tacgagaaga ccggtcttgc tagattctaa tcaagaggat gtcagaatgc 12840

catttgcctg agagatgcag gcttcatttt tgatactttt ttatttgtaa cctatatagt 12900

ataggatttt ttttgtcatt ttgtttcttc tcgtacgagc ttgctcctga tcagcctatc 12960

tcgcagctga tgaatatctt gtggtagggg tttgggaaaa tcattcgagt ttgatgtttt 13020

tcttggtatt tcccactcct cttcagagta cagaagatta agtgagacct tcgtttgtgc 13080

ggatcctaat gcggtagttt atcacagtta aattgctaac gcagtcaggc accgtgtatg 13140

aaatctaaca atgcgctcat cgtcatcctc ggcaccgtca ccctggatgc tgtaggcata 13200

ggcttggtta tgccggtact gccgggcctc ttgcgggata tcgtccattc cgacagcatc 13260

gccagtcact atggcgtgct gctagcgcta tatgcgttga tgcaatttct atgcgcaccc 13320

gttctcggag cactgtccga ccgctttggc cgccgcccag tcctgctcgc ttcgctactt 13380

ggagccacta tcgactacgc gatcatggcg accacacccg tcctgtggat ctatcgaatc 13440

taaatgtaag ttaaaatctc taaataatta aataagtccc agtttctcca tacgaacctt 13500

aacagcattg cggtgagcat ctagaccttc aacagcagcc agatccatca ctgcttggcc 13560

aatatgtttc agtccctcag gagttacgtc ttgtgaagtg atgaacttct ggaaggttgc 13620

agtgttaact ccgctgtatt gacgggcata tccgtacgtt ggcaaagtgt ggttggtacc 13680

ggaggagtaa tctccacaac tctctggaga gtaggcacca acaaacacag atccagcgtg 13740

ttgtacttga tcaacataag aagaagcatt ctcgatttgc aggatcaagt gttcaggagc 13800

gtactgattg gacatttcca aagcctgctc gtaggttgca accgataggg ttgtagagtg 13860

tgcaatacac ttgcgtacaa tttcaaccct tggcaactgc acagcttggt tgtgaacagc 13920

atcttcaatt ctggcaagct ccttgtctgt catatcgaca gccaacagaa tcacctggga 13980

atcaatacca tgttcagctt gagacagaag gtctgaggca acgaaatctg gatcagcgta 14040

tttatcagca ataactagaa cttcagaagg cccagcaggc atgtcaatac tacacagggc 14100

tgatgtgtca ttttgaacca tcatcttggc agcagtaacg aactggtttc ctggaccaaa 14160

tattttgtca cacttaggaa cagtttctgt tccgtaagcc atagcagcta ctgcctgggc 14220

gcctcctgct agcacgatac acttagcacc aaccttgtgg gcaacgtaga tgacttctgg 14280

ggtaagggta ccatccttct taggtggaga tgcaaaaaca atttctttgc aaccagcaac 14340

tttggcagga acacccagca tcagggaagt ggaaggcaga attgcggttc caccaggaat 14400

atagaggcca actttctcaa taggtcttgc aaaacgagag cagactacac cagggcaagt 14460

ctcaacttgc aacgtctccg ttagttgagc ttcatggaat ttcctgacgt tatctataga 14520

gagatcaatg gctctcttaa cgttatctgg caattgcata agttcctctg ggaaaggagc 14580

ttctaacaca ggtgtcttca aagcgactcc atcaaacttg gcagttagtt ctaaaagggc 14640

tttgtcacca ttttgacgaa cattgtcgac aattggtttg actaattcca taatctgttc 14700

cgttttctgg ataggacgac gaagggcatc ttcaatttct tgtgaggagg ccttagaaac 14760

gtcaattttg cacaattcaa tacgaccttc agaagggact tctttaggtt tggattcttc 14820

tttaggttgt tccttggtgt atcctggctt ggcatctcct ttccttctag tgacctttag 14880

ggacttcata tccaggtttc tctccacctc gtccaacgtc acaccgtact tggcacatct 14940

aactaatgca aaataaaata agtcagcaca ttcccaggct atatcttcct tggatttagc 15000

ttctgcaagt tcatcagctt cctccctaat tttagcgttc aacaaaactt cgtcgtcaaa 15060

taaccgtttg gtataagaac cttctggagc attgctctta cgatcccaca aggtggcttc 15120

catggctcta agaccctttg attggccaaa acaggaagtg cgttccaagt gacagaaacc 15180

aacacctgtt tgttcaacca caaatttcaa gcagtctcca tcacaatcca attcgatacc 15240

cagcaacttt tgagttgctc cagatgtagc acctttatac cacaaaccgt gacgacgaga 15300

ttggtagact ccagtttgtg tccttatagc ctccggaata gactttttgg acgagtacac 15360

caggcccaac gagtaattag aagagtcagc caccaaagta gtgaatagac catcggggcg 15420

gtcagtagtc aaagacgcca acaaaatttc actgacaggg aactttttga catcttcaga 15480

aagttcgtat tcagtagtca attgccgagc atcaataatg gggattatac cagaagcaac 15540

agtggaagtc acatctacca actttgcggt ctcagaaaaa gcataaacag ttctactacc 15600

gccattagtg aaacttttca aatcgcccag tggagaagaa aaaggcacag cgatactagc 15660

attagcgggc aaggatgcaa ctttatcaac cagggtccta tagataaccc tagcgcctgg 15720

gatcatcctt tggacaactc tttctgccaa atctaggtcc aaaatcactt cattgatacc 15780

attattgtac aacttgagca agttgtcgat cagctcctca aattggtcct ctgtaacgga 15840

tgactcaact tgcacattaa cttgaagctc agtcgattga gtgaacttga tcaggttgtg 15900

cagctggtca gcagcatagg gaaacacggc ttttcctacc aaactcaagg aattatcaaa 15960

ctctgcaaca cttgcgtatg caggtagcaa gggaaatgtc atacttgaag tcggacagtg 16020

agtgtagtct tgagaaattc tgaagccgta tttttattat cagtgagtca gtcatcagga 16080

gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg 16140

cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag 16200

cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat 16260

ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg 16320

ctgcttccta atgcaggagt cgcataaggg agagcgtcga gtatctatga ttggaagtat 16380

gggaatggtg atacccgcat tcttcagtgt cttgaggtct cctatcagat tatgcccaac 16440

taaagcaacc ggaggaggag atttcatggt aaatttctct gacttttggt catcagtaga 16500

ctcgaactgt gagactatct cggttatgac agcagaaatg tccttcttgg agacagtaaa 16560

tgaagtccca ccaataaaga aatccttgtt atcaggaaca aacttcttgt ttcgaacttt 16620

ttcggtgcct tgaactataa aatgtagagt ggatatgtcg ggtaggaatg gagcgggcaa 16680

atgcttacct tctggacctt caagaggtat gtagggtttg tagatactga tgccaacttc 16740

agtgacaacg ttgctatttc gttcaaacca ttccgaatcc agagaaatca aagttgtttg 16800

tctactattg atccaagcca gtgcggtctt gaaactgaca atagtgtgct cgtgttttga 16860

ggtcatcttt gtatgaataa atctagtctt tgatctaaat aatcttgacg agccaaggcg 16920

ataaataccc aaatctaaaa ctcttttaaa acgttaaaag gacaagtatg tctgcctgta 16980

ttaaacccca aatcagctcg tagtctgatc ctcatcaact tgaggggcac tatcttgttt 17040

tagagaaatt tgcggagatg cgatatcgag aaaaaggtac gctgatttta aacgtgaaat 17100

ttatctcaag atctgctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 17160

gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 17220

tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag 17280

cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg 17340

caccatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc 17400

tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 17460

tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 17520

aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 17580

tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 17640

tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 17700

cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 17760

agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 17820

tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 17880

aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 17940

ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 18000

cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt 18060

accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 18120

ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 18180

ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 18240

gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 18300

aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 18360

gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 18420

gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 18480

cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 18540

gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 18600

gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctgca 18660

ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 18720

tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 18780

ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 18840

cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 18900

accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca 18960

cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 19020

tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 19080

cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 19140

acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 19200

atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 19260

tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 19320

aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 19380

cgtatcacga ggccctttcg tcttcaagaa ttaattctca tgtttgacag cttatcatcg 19440

ataagctgac tcatgttggt attgtgaaat agacgcagat cgggaacact gaaaaataac 19500

agttattatt cg 19512

<210> 5

<211> 7108

<212> DNA

<213> unknown (Artificial Synthesis)

<400> 5

agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 60

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 120

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 180

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 240

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 300

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 360

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 420

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 480

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 540

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 600

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 660

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 720

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 780

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 840

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 900

caacttgaga agatcaaaaa acaactaatt attcgaaacg aggaattcac gtggcccagc 960

cggccgtctc ggatcggtac ctcgagatgg gtaaatcaat tggaattgat ttgggtacca 1020

catactcttg tgtggcacat tttgctaatg atcgtgttga gatcatagct aacgaccaag 1080

gtaacaggac gactccatcg ttcgtcgcct ttaccgacac tgaaagattg attggtgatg 1140

ctgcaaagaa ccaagctgcc atgaatccag ctaacactgt tttcgatgcc aaacgtttaa 1200

tcggtagaaa attcgacgac ccggaaactc aggccgatat taagcacttc cccttcaaag 1260

ttatcaacaa ggggggaaag cctaatatcc aagtcgaatt taagggtgag actaaggttt 1320

tcagccccga agagatttcc tccatggttc taacaaaaat gaaggatact gctgagcagt 1380

atttgggtga gaaaatcaac gatgcagttg tcactgttcc tgcttacttc aatgactctc 1440

aaagacaagc caccaaggat gctggtttga ttgctggttt gaacgttcaa agaatcatta 1500

atgagcccac cgctgccgca attgcttacg ggttggacaa gaaggatgca ggccacggtg 1560

agcacaacat tctaatcttc gatctaggtg gaggaacttt cgatgtttct ctactatcta 1620

ttgatgaggg tattttcgaa gtcaaggcca ccgcaggtga cacccacttg ggtggtgagg 1680

acttcgataa cagattagtc aaccacttta tcgccgagtt caagagaaag accaagaaag 1740

atctttctac aaaccagaga tcccttagaa gactaagaac cgcttgtgag cgtgcaaaga 1800

gaactttgtc ttcttctgct cagacctcca tcgagattga ttctttgttc gagggtatcg 1860

acttctacac ctcgatcact agagctagat tcgaggagct ctgtgccgac ttgttcagat 1920

ccaccatcga gcctgttgag agagtcttga aagactccaa gttggacaaa tctcaagttc 1980

atgagattgt tttggttggt ggttctacca gaattccaaa ggttcagaaa ttagtttctg 2040

actttttcaa tggtaaggag ccaaacaagt ccatcaaccc agacgaagcc gttgcatatg 2100

gtgctgctgt ccaagcagct attttgtctg gagatacttc ttccaagaca caagacttgt 2160

tattgctgga tgttgctcct ctatctttgg gtattgaaac cgctggtggt atcatgacca 2220

agctgatccc aagaaactcc acaatcccag ccaaaaagtc agaaatcttt tcgacatatg 2280

ctgacaacca accaggtgtt ttgattcaag tctttgaagg tgagagaact agaaccaagg 2340

acaacaacct gttgggtaag tttgaacttt ctggtattcc tcctgctcca agaggtgttc 2400

ctcaaattga ggtcaccttc gatatggatg ccaacggtat tttgaatgta tctgctgttg 2460

agaagggtac cggtaagact caaaagatta ctattaccaa cgataaggga agattgtcca 2520

aggaagacat cgagagaatg gtttctgaag ctgaaaaatt caaggatgaa gacgagaagg 2580

aagccgagag agttgctgcc aagaatggct tggaatcata tgcttactct ctgaagaact 2640

ctgcagctga atctggattc aaggacaagg ttggagagga tgatcttgcc aagttgaaca 2700

agtcagttga agagacaata tcttggttag atgagtcaca atctgcttcc acagacgagt 2760

acaaggacag gcaaaaggaa ttggaagaag ttgctaaccc aataatgagc aagttctatg 2820

gagctgctgg tggagctcct ggtggagctc ctggtggctt ccctggaggt ttccctggcg 2880

gagctggcgc agctggcggt gccccaggtg gtgctgcccc aggcggagac agcggaccaa 2940

ccgtggaaga agtcgattaa gcggccgcca gcttacgtag aacaaaaact catctcagaa 3000

gaggatctga atagcgccgt cgaccatcat catcatcatc attgagtttg tagccttaga 3060

catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg ctagattcta 3120

atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt ttgatacttt 3180

tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt ctcgtacgag 3240

cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg gtttgggaaa 3300

atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt acagaagatt 3360

aagtgagacc ttcgtttgtg cggatctaac atccaaagac gaaaggttga atgaaacctt 3420

tttgccatcc gacatccaca ggtccattct cacacataag tgccaaacgc aacaggaggg 3480

gatacactag cagcagaccg ttgcaaacgc aggacctcca ctcctcttct cctcaacacc 3540

cacttttgcc atcgaaaaac cagcccagtt attgggcttg attggagctc gctcattcca 3600

attccttcta ttaggctact aacaccatga ctttattagc ctgtctatcc tggcccccct 3660

ggcgaggttc atgtttgttt atttccgaat gcaacaagct ccgcattaca cccgaacatc 3720

actccagatg agggctttct gagtgtgggg tcaaatagtt tcatgttccc caaatggccc 3780

aaaactgaca gtttaaacgc tgtcttggaa cctaatatga caaaagcgtg atctcatcca 3840

agatgaacta agtttggttc gttgaaatgc taacggccag ttggtcaaaa agaaacttcc 3900

aaaagtcggc ataccgtttg tcttgtttgg tattgattga cgaatgctca aaaataatct 3960

cattaatgct tagcgcagtc tctctatcgc ttctgaaccc cggtgcacct gtgccgaaac 4020

gcaaatgggg aaacacccgc tttttggatg attatgcatt gtctccacat tgtatgcttc 4080

caagattctg gtgggaatac tgctgatagc ctaacgttca tgatcaaaat ttaactgttc 4140

taacccctac ttgacagcaa tatataaaca gaaggaagct gccctgtctt aaaccttttt 4200

ttttatcatc attattagct tactttcata attgcgactg gttccaattg acaagctttt 4260

gattttaacg acttttaacg acaacttgag aagatcaaaa aacaactaat tattcgaaac 4320

gaggaattca tgttagacca gcaaaccatt aacatcatca aagccactgt tcctgtattg 4380

aaggagcatg gcgttaccat taccacgact ttttataaaa acttgtttgc caaacaccct 4440

gaagtacgtc ctttgtttga tatgggtcgc caagaatctt tggagcagcc taaggctttg 4500

gcgatgacgg tattggcggc agcgcaaaac attgaaaatt tgccagctat tttgcctgcg 4560

gtcaaaaaaa ttgcagtcaa acattgtcaa gcaggcgtgg cagcagcgca ttatccgatt 4620

gtcggtcaag aattgttggg tgcgattaaa gaagtattgg gcgatgccgc aaccgatgac 4680

attttggacg cgtggggcaa ggcttatggc gtgattgcag atgtgtttat tcaagtggaa 4740

gcagatttgt acgctcaagc ggttgaataa gcggccgcca gcttacgtag aacaaaaact 4800

catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagtttg 4860

tagccttaga catgactgtt cctcagttca agttgggcac ttacgagaag accggtcttg 4920

ctagattcta atcaagagga tgtcagaatg ccatttgcct gagagatgca ggcttcattt 4980

ttgatacttt tttatttgta acctatatag tataggattt tttttgtcat tttgtttctt 5040

ctcgtacgag cttgctcctg atcagcctat ctcgcagctg atgaatatct tgtggtaggg 5100

gtttgggaaa atcattcgag tttgatgttt ttcttggtat ttcccactcc tcttcagagt 5160

acagaagatt aagtgagacc ttcgtttgtg cggatccccc acacaccata gcttcaaaat 5220

gtttctactc cttttttact cttccagatt ttctcggact ccgcgcatcg ccgtaccact 5280

tcaaaacacc caagcacagc atactaaatt ttccctcttt cttcctctag ggtgtcgtta 5340

attacccgta ctaaaggttt ggaaaagaaa aaagagaccg cctcgtttct ttttcttcgt 5400

cgaaaaaggc aataaaaatt tttatcacgt ttctttttct tgaaattttt ttttttagtt 5460

tttttctctt tcagtgacct ccattgatat ttaagttaat aaacggtctt caatttctca 5520

agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgt tcattagaaa 5580

gaaagcatag caatctaatc taaggggcgg tgttgacaat taatcatcgg catagtatat 5640

cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga ccagtgccgt 5700

tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg accggctcgg 5760

gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg acgtgaccct 5820

gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct gggtgtgggt 5880

gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga acttccggga 5940

cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg agttcgccct 6000

gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact gacacgtccg 6060

acggcggccc acgggtccca ggcctcggag atccgtcccc cttttccttt gtcgatatca 6120

tgtaattagt tatgtcacgc ttacattcac gccctccccc cacatccgct ctaaccgaaa 6180

aggaaggagt tagacaacct gaagtctagg tccctattta tttttttata gttatgttag 6240

tattaagaac gttatttata tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg 6300

catgtaacat tatactgaaa accttgcttg agaaggtttt gggacgctcg aaggctttaa 6360

tttgcaagct ggagaccaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 6420

aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 6480

gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 6540

ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 6600

cctttctccc ttcgggaagc gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt 6660

cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 6720

gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 6780

cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 6840

agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg 6900

ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 6960

ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 7020

gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 7080

cacgttaagg gattttggtc atgagatc 7108

<210> 6

<211> 11903

<212> DNA

<213> Unknown (Artificial Synthesis)

<400> 6

agatctaaca tccaaagacg aaaggttgaa tgaaaccttt ttgccatccg acatccacag 60

gtccattctc acacataagt gccaaacgca acaggagggg atacactagc agcagaccgt 120

tgcaaacgca ggacctccac tcctcttctc ctcaacaccc acttttgcca tcgaaaaacc 180

agcccagtta ttgggcttga ttggagctcg ctcattccaa ttccttctat taggctacta 240

acaccatgac tttattagcc tgtctatcct ggcccccctg gcgaggttca tgtttgttta 300

tttccgaatg caacaagctc cgcattacac ccgaacatca ctccagatga gggctttctg 360

agtgtggggt caaatagttt catgttcccc aaatggccca aaactgacag tttaaacgct 420

gtcttggaac ctaatatgac aaaagcgtga tctcatccaa gatgaactaa gtttggttcg 480

ttgaaatgct aacggccagt tggtcaaaaa gaaacttcca aaagtcggca taccgtttgt 540

cttgtttggt attgattgac gaatgctcaa aaataatctc attaatgctt agcgcagtct 600

ctctatcgct tctgaacccc ggtgcacctg tgccgaaacg caaatgggga aacacccgct 660

ttttggatga ttatgcattg tctccacatt gtatgcttcc aagattctgg tgggaatact 720

gctgatagcc taacgttcat gatcaaaatt taactgttct aacccctact tgacagcaat 780

atataaacag aaggaagctg ccctgtctta aacctttttt tttatcatca ttattagctt 840

actttcataa ttgcgactgg ttccaattga caagcttttg attttaacga cttttaacga 900

caacttgaga agatcaaaaa acaactaatt attcgaaacg aggaattcat gagtaaccag 960

tataatccgt atgagcagaa ccagtcttac gagctgccct cgtacaaggg gggcaacaac 1020

gacgattttg tcaagtttat gaacgaaatt gctgacatca acgccaattt ggacaactac 1080

gaagaactag tgaagattat tgagcaaaaa caaacccaac tcgtgaatga agtcaaccca 1140

gaccaggaga actctttgaa acgacagcta gactccctaa tcagtgaatc ctcctctttg 1200

caattatcgt taaagtccaa gattaagaac gctcaacagc tggctattgg cgactcggct 1260

aaggtgggac aagctgagac cagtcgtcaa cgttttcttc aagctattca ggactaccgt 1320

attattgaaa gcaactacag agaacagcag cgagtccaag ccgaaaggca atatcgtgtc 1380

gttaagcccg atgcctcccc tgaagaggtc agagatgcta tcgacgactt gggtggccag 1440

caggtgttct caacggcttt gctgaacgct aataggagag gagaggcaaa gacggcgctt 1500

caagaagttc aatctcgtca cagagagttg caaaggttag aaaagactat ggctgaactt 1560

actcaactgt tccatgacat ggaggagatg gtagttgagc aggatcaaca cgttcaagag 1620

accgagaact tggtagacac tgcccagcag gacattgaaa aagcagtcgg ccacaccgat 1680

aaggccttga ctagtgctaa aaaggcacgg agaaagaagt gtatctgctt ctggatctgt 1740

gtccttatca tctgtattct tgcactgatc cttggtctgg gattcggtgt cggaaactgg 1800

ggaagatagg cggccgcgtt tgtagcctta gacatgactg ttcctcagtt caagttgggc 1860

acttacgaga agaccggtct tgctagattc taatcaagag gatgtcagaa tgccatttgc 1920

ctgagagatg caggcttcat ttttgatact tttttatttg taacctatat agtataggat 1980

tttttttgtc attttgtttc ttctcgtacg agcttgctcc tgatcagcct atctcgcagc 2040

tgatgaatat cttgtggtag gggtttggga aaatcattcg agtttgatgt ttttcttggt 2100

atttcccact cctcttcaga gtacagaaga ttaagtgaga cgttcgtttg tgcggatcta 2160

acatccaaag acgaaaggtt gaatgaaacc tttttgccat ccgacatcca caggtccatt 2220

ctcacacata agtgccaaac gcaacaggag gggatacact agcagcagac cgttgcaaac 2280

gcaggacctc cactcctctt ctcctcaaca cccacttttg ccatcgaaaa accagcccag 2340

ttattgggct tgattggagc tcgctcattc caattccttc tattaggcta ctaacaccat 2400

gactttatta gcctgtctat cctggccccc ctggcgaggt tcatgtttgt ttatttccga 2460

atgcaacaag ctccgcatta cacccgaaca tcactccaga tgagggcttt ctgagtgtgg 2520

ggtcaaatag tttcatgttc cccaaatggc ccaaaactga cagtttaaac gctgtcttgg 2580

aacctaatat gacaaaagcg tgatctcatc caagatgaac taagtttggt tcgttgaaat 2640

gctaacggcc agttggtcaa aaagaaactt ccaaaagtcg gcataccgtt tgtcttgttt 2700

ggtattgatt gacgaatgct caaaaataat ctcattaatg cttagcgcag tctctctatc 2760

gcttctgaac cccggtgcac ctgtgccgaa acgcaaatgg ggaaacaccc gctttttgga 2820

tgattatgca ttgtctccac attgtatgct tccaagattc tggtgggaat actgctgata 2880

gcctaacgtt catgatcaaa atttaactgt tctaacccct acttgacagc aatatataaa 2940

cagaaggaag ctgccctgtc ttaaaccttt ttttttatca tcattattag cttactttca 3000

taattgcgac tggttccaat tgacaagctt ttgattttaa cgacttttaa cgacaacttg 3060

agaagatcaa aaaacaacta attattcgaa acgaggaatt catgtcaaga gaagattctg 3120

tttatttagc aaaactagct gagcaagctg agcgttatga ggagatggtc gagaacatga 3180

agaccgtcgc ctcttccggc ttagagttgt ctgtcgaaga gagaaacttg ctttctgttg 3240

catacaaaaa cgtaattgga gctagaagag cttcttggag aatcgtctcc tcaattgaac 3300

agaaagagga agccaagggt aaccaatcac aagtgtcttt gatcagagaa taccgctcca 3360

agattgagac cgaattggcc aacatttgtg aggatatttt gtctgttttg agtgagcacc 3420

ttattccttc tgccagaact ggcgaatcca aggtcttcta ctttaagatg aagggtgatt 3480

accaccgtta tttggccgaa tttgctgttg gtgacaagcg aaaggaagct gctaatttgt 3540

cattggaggc ttacaagtct gcctctgacg ttgctgttac ggagctacct ccaactcatc 3600

caattagatt gggtctggct ctgaatttct cagtcttcta ctacgagatt ctaaactctc 3660

ctgaccgcgc ctgtcattta gccaagcaag ctttcgacga tgctattgct gagttagaaa 3720

ccctatctga agaatcttac aaagactcca ctttgattat gcaactgctg cgtgacaact 3780

tgactttgtg gacctcagac atgtctgaaa ctggacaaga agagtcatcc aatagccaag 3840

ataagacaga agctgctccc aaagatgaag agtgagcggc cgcgtttgta gccttagaca 3900

tgactgttcc tcagttcaag ttgggcactt acgagaagac cggtcttgct agattctaat 3960

caagaggatg tcagaatgcc atttgcctga gagatgcagg cttcattttt gatacttttt 4020

tatttgtaac ctatatagta taggattttt tttgtcattt tgtttcttct cgtacgagct 4080

tgctcctgat cagcctatct cgcagctgat gaatatcttg tggtaggggt ttgggaaaat 4140

cattcgagtt tgatgttttt cttggtattt cccactcctc ttcagagtac agaagattaa 4200

gtgagacgtt cgtttgtgcg gatcctaatg cggtagttta tcacagttaa attgctaacg 4260

cagtcaggca ccgtgtatga aatctaacaa tgcgctcatc gtcatcctcg gcaccgtcac 4320

cctggatgct gtaggcatag gcttggttat gccggtactg ccgggcctct tgcgggatat 4380

cgtccattcc gacagcatcg ccagtcacta tggcgtgctg ctagcgctat atgcgttgat 4440

gcaatttcta tgcgcacccg ttctcggagc actgtccgac cgctttggcc gccgcccagt 4500

cctgctcgct tcgctacttg gagccactat cgactacgcg atcatggcga ccacacccgt 4560

cctgtggatc tatcgaatct aaatgtaagt taaaatctct aaataattaa ataagtccca 4620

gtttctccat acgaacctta acagcattgc ggtgagcatc tagaccttca acagcagcca 4680

gatccatcac tgcttggcca atatgtttca gtccctcagg agttacgtct tgtgaagtga 4740

tgaacttctg gaaggttgca gtgttaactc cgctgtattg acgggcatat ccgtacgttg 4800

gcaaagtgtg gttggtaccg gaggagtaat ctccacaact ctctggagag taggcaccaa 4860

caaacacaga tccagcgtgt tgtacttgat caacataaga agaagcattc tcgatttgca 4920

ggatcaagtg ttcaggagcg tactgattgg acatttccaa agcctgctcg taggttgcaa 4980

ccgatagggt tgtagagtgt gcaatacact tgcgtacaat ttcaaccctt ggcaactgca 5040

cagcttggtt gtgaacagca tcttcaattc tggcaagctc cttgtctgtc atatcgacag 5100

ccaacagaat cacctgggaa tcaataccat gttcagcttg agacagaagg tctgaggcaa 5160

cgaaatctgg atcagcgtat ttatcagcaa taactagaac ttcagaaggc ccagcaggca 5220

tgtcaatact acacagggct gatgtgtcat tttgaaccat catcttggca gcagtaacga 5280

actggtttcc tggaccaaat attttgtcac acttaggaac agtttctgtt ccgtaagcca 5340

tagcagctac tgcctgggcg cctcctgcta gcacgataca cttagcacca accttgtggg 5400

caacgtagat gacttctggg gtaagggtac catccttctt aggtggagat gcaaaaacaa 5460

tttctttgca accagcaact ttggcaggaa cacccagcat cagggaagtg gaaggcagaa 5520

ttgcggttcc accaggaata tagaggccaa ctttctcaat aggtcttgca aaacgagagc 5580

agactacacc agggcaagtc tcaacttgca acgtctccgt tagttgagct tcatggaatt 5640

tcctgacgtt atctatagag agatcaatgg ctctcttaac gttatctggc aattgcataa 5700

gttcctctgg gaaaggagct tctaacacag gtgtcttcaa agcgactcca tcaaacttgg 5760

cagttagttc taaaagggct ttgtcaccat tttgacgaac attgtcgaca attggtttga 5820

ctaattccat aatctgttcc gttttctgga taggacgacg aagggcatct tcaatttctt 5880

gtgaggaggc cttagaaacg tcaattttgc acaattcaat acgaccttca gaagggactt 5940

ctttaggttt ggattcttct ttaggttgtt ccttggtgta tcctggcttg gcatctcctt 6000

tccttctagt gacctttagg gacttcatat ccaggtttct ctccacctcg tccaacgtca 6060

caccgtactt ggcacatcta actaatgcaa aataaaataa gtcagcacat tcccaggcta 6120

tatcttcctt ggatttagct tctgcaagtt catcagcttc ctccctaatt ttagcgttca 6180

acaaaacttc gtcgtcaaat aaccgtttgg tataagaacc ttctggagca ttgctcttac 6240

gatcccacaa ggtggcttcc atggctctaa gaccctttga ttggccaaaa caggaagtgc 6300

gttccaagtg acagaaacca acacctgttt gttcaaccac aaatttcaag cagtctccat 6360

cacaatccaa ttcgataccc agcaactttt gagttgctcc agatgtagca cctttatacc 6420

acaaaccgtg acgacgagat tggtagactc cagtttgtgt ccttatagcc tccggaatag 6480

actttttgga cgagtacacc aggcccaacg agtaattaga agagtcagcc accaaagtag 6540

tgaatagacc atcggggcgg tcagtagtca aagacgccaa caaaatttca ctgacaggga 6600

actttttgac atcttcagaa agttcgtatt cagtagtcaa ttgccgagca tcaataatgg 6660

ggattatacc agaagcaaca gtggaagtca catctaccaa ctttgcggtc tcagaaaaag 6720

cataaacagt tctactaccg ccattagtga aacttttcaa atcgcccagt ggagaagaaa 6780

aaggcacagc gatactagca ttagcgggca aggatgcaac tttatcaacc agggtcctat 6840

agataaccct agcgcctggg atcatccttt ggacaactct ttctgccaaa tctaggtcca 6900

aaatcacttc attgatacca ttattgtaca acttgagcaa gttgtcgatc agctcctcaa 6960

attggtcctc tgtaacggat gactcaactt gcacattaac ttgaagctca gtcgattgag 7020

tgaacttgat caggttgtgc agctggtcag cagcataggg aaacacggct tttcctacca 7080

aactcaagga attatcaaac tctgcaacac ttgcgtatgc aggtagcaag ggaaatgtca 7140

tacttgaagt cggacagtga gtgtagtctt gagaaattct gaagccgtat ttttattatc 7200

agtgagtcag tcatcaggag atcctctacg ccggacgcat cgtggccgac ctgcaggggg 7260

ggggggggcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac caggcctgaa 7320

tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct ttgttgtagg 7380

tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg ttgtcgggaa 7440

gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa agccgccgtc 7500

ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt ctgattagaa 7560

aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata 7620

tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat 7680

ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa 7740

tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc 7800

cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt 7860

acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg 7920

agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa 7980

ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc 8040

taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg 8100

agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct 8160

gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc 8220

tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc 8280

gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcctcga 8340

gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc 8400

agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt 8460

ttgagacaca acgtggcttt cccccccccc cctgcaggtc ggcatcaccg gcgccacagg 8520

tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt 8580

cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact 8640

gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa 8700

cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagtatctat 8760

gattggaagt atgggaatgg tgatacccgc attcttcagt gtcttgaggt ctcctatcag 8820

attatgccca actaaagcaa ccggaggagg agatttcatg gtaaatttct ctgacttttg 8880

gtcatcagta gactcgaact gtgagactat ctcggttatg acagcagaaa tgtccttctt 8940

ggagacagta aatgaagtcc caccaataaa gaaatccttg ttatcaggaa caaacttctt 9000

gtttcgaact ttttcggtgc cttgaactat aaaatgtaga gtggatatgt cgggtaggaa 9060

tggagcgggc aaatgcttac cttctggacc ttcaagaggt atgtagggtt tgtagatact 9120

gatgccaact tcagtgacaa cgttgctatt tcgttcaaac cattccgaat ccagagaaat 9180

caaagttgtt tgtctactat tgatccaagc cagtgcggtc ttgaaactga caatagtgtg 9240

ctcgtgtttt gaggtcatct ttgtatgaat aaatctagtc tttgatctaa ataatcttga 9300

cgagccaagg cgataaatac ccaaatctaa aactctttta aaacgttaaa aggacaagta 9360

tgtctgcctg tattaaaccc caaatcagct cgtagtctga tcctcatcaa cttgaggggc 9420

actatcttgt tttagagaaa tttgcggaga tgcgatatcg agaaaaaggt acgctgattt 9480

taaacgtgaa atttatctca agatctctgc ctcgcgcgtt tcggtgatga cggtgaaaac 9540

ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc 9600

agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc 9660

cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg 9720

tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 9780

gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 9840

ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 9900

acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 9960

cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 10020

caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 10080

gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 10140

tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 10200

aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 10260

ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 10320

cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 10380

tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 10440

tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 10500

ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 10560

aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 10620

aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 10680

aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 10740

gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 10800

gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 10860

caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 10920

ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 10980

attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 11040

ccattgctgc aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 11100

gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 11160

ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 11220

tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 11280

gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 11340

cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg 11400

gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga 11460

tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg 11520

ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat 11580

gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc 11640

tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca 11700

catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct 11760

ataaaaatag gcgtatcacg aggccctttc gtcttcaaga attaattctc atgtttgaca 11820

gcttatcatc gataagctga ctcatgttgg tattgtgaaa tagacgcaga tcgggaacac 11880

tgaaaaataa cagttattat tcg 11903

<210> 7

<211> 540

<212> DNA

<213> Unknown (Artificial Synthesis)

<400> 7

gttccacctc catgtgattt gggtattgct tctaaggtta aacaaaaggg tgttactggt 60

ggtggtgctt ctgtttctac tacttctgct actcaaggtt ctggtactac taactgtgtt 120

actagaactc ctaattctgt tgaaaagaaa aacgttgctg gtaatactgg tgttactgct 180

acttctgttt ctgctggaga tggtgctttt ggtaacttgg ctgctgcttt gactttggtt 240

gaagatactg aggatggttt gggtgttaaa actaagaacg gtggtaaagg tttctctgag 300

ggtactgctg ctatttctca aactgctggt gctaatggtg gtgctactgt taagaaagca 360

aagttggatt tgttgactga tggtgaagat ttgttcgata ctaagaaagt tgagaagggt 420

actgttactt cttcttcttc tcatcaaggt tctggtgctg gagattctat cttcgaaatc 480

ttgaacgaag ctgagtctaa gattaagaaa tctggagatc atcaccatca ccatcactaa 540

<210> 8

<211> 570

<212> DNA

<213> unknown (Artificial Synthesis)

<400> 8

catgaagagg atggtgtttg taactctaat gctccatgtt accactgtga tgctaacggt 60

gaaaactgtt cttgtaactg tgaattgttt gattgtgagg ctaagaaacc agatggttct 120

tatgctcatc cttgtagaag atgtgatgct aacaacatct gtaagtgttc ttgtactgct 180

atcccatgta atgaagatca tccttgtcat cactgtcacg aagaggatga tggagatact 240

cattgtcact gttcttgtga gcattctcac gatcatcacg atgatgatac tcatggtgaa 300

tgtactaaga aagctccatg ttggcgttgt gagtacaacg ctgatttgaa gcatgatgtt 360

tgtggttgtg aatgttctaa attgccatgt aatgatgaac acccttgtta tagaaaggag 420

ggtggtgttg tttcttgtga ttgtaagact atcacttgta acgaggatca tccttgttac 480

cactcttatg aagaggatgg tgttactaag tctgattgtg attgtgaaca ttctccaggt 540

ccttctgagc atcaccatca ccatcactaa 570

<210> 9

<211> 1683

<212> DNA

<213> unknown (Artificial Synthesis)

<400> 9

gtccccgtgt ccggaaagag cggcagctcg actaccgccg tgtcagccag cgataattcg 60

gcgctcccgc cactgatatc gtctcggtgc gcgcctccgt ccaacaaagg ttccaagtct 120

gatttacagg ctgagcccta ctacatgcag aaaaacaccg aatggtacga gagccatggc 180

ggcaacctaa caagcatagg taagagggat gacaatttgg tggggggtat gacgcttgac 240

ttgccgtcgg atgctcctcc tatatcgctc agtggctcta ccaatagtgc ctcggatggc 300

ggaaaagtag ttgcggcgac gacggcgcag atccaagaat ttactaagta tgcagggata 360

gccgcaactg catactgccg cagtgtcgtg cctggcaaca aatgggactg cgttcagtgc 420

cagaagtggg tacccgacgg caagattatt actactttca catcactgtt gtctgacaca 480

aatggctatg ttttacgatc agacaagcaa aaaacaatct atttagtctt cagaggaact 540

aatagctttc gttcagctat aacggatatt gttttcaact ttagtgacta caaacccgtt 600

aaaggagcca aggtccatgc tgggtttcta tcgtcttacg agcaagttgt aaacgattac 660

ttccctgtcg tgcaagagca gctgactgca aacccgacat ataaagtaat tgtcacgggt 720

cactctctgg gaggggccca ggcattactg gctgggatgg atttatatca aagggaacca 780

cggctttctc caaagaactt aagcattttt accgtaggcg gcccacgtgt ggggaacccc 840

acttttgctt attatgtcga gtctacaggg attccgtttc agagaactgt acacaagcga 900

gatatagtac ctcatgtacc accccaatcc ttcggattcc ttcatccagg ggtggagtct 960

tggatcaaaa gtggtactag taatgtccaa atttgtacgt ccgaaataga aaccaaagac 1020

tgctctaatt ctatagttcc ctttacgagt atccttgatc acctatcata tttcgacatc 1080

aatgagggat cttgtctagg cgggggtggg tctggaggtg gtggaagtgg cggtggtggc 1140

agtgttccgc ctccatgtga cctgggtatc gccagcaagg ttaaacaaaa aggcgtaaca 1200

gggggaggtg catccgtgtc aacgacctct gcgactcaag gttctggtac aactaattgt 1260

gtcacgcgca ccccgaactc ggtagagaaa aagaatgtcg cgggcaatac gggggttaca 1320

gctaccagtg tgtcagcggg cgatggagct ttcggaaatt tggctgctgc actcacacta 1380

gtcgaagata cagaggacgg tctcggtgta aagaccaaaa atgggggaaa gggtttttcc 1440

gaagggactg ccgcgattag ccaaacggca ggagcaaacg ggggggccac ggtaaaaaag 1500

gcgaaacttg acttgttgac tgatggagaa gaccttttcg acaccaagaa agttgagaag 1560

ggaaccgtga cctccagttc ctcgcaccaa ggctcaggtg caggggacag catttttgag 1620

atcctcaacg aagccgaatc gaaaatcaag aagtctggtg atcaccacca ccatcatcat 1680

tga 1683

<210> 10

<211> 1713

<212> DNA

<213> Unknown (Artificial Synthesis)

<400> 10

gtcccagtct ccggtaagtc tgggtcaagt actacagcag tcagtgcttc ggacaactcc 60

gcgcttccgc cgctgattag ttcgcgatgt gctccgccct ctaacaaggg tagtaaatct 120

gacttacagg cggagccata ttacatgcag aaaaacaccg agtggtatga gagccacggc 180

ggtaacttga ccagtatcgg gaagcgcgat gacaacttag ttggggggat gacactggat 240

ctgcctagcg atgcaccccc catatctttg tcgggatcga ctaattctgc aagcgacggt 300

ggaaaggtag ttgcagctac gacggctcaa atccaggagt tcacaaaata cgccggaata 360

gcggccacgg cttattgtcg gtctgtagtc ccagggaata agtgggactg cgtccagtgc 420

cagaaatggg tacctgacgg taagattatc actacattta ccagtttact gtcggatacg 480

aatgggtacg ttctaaggag cgacaaacaa aaaaccatct atctagtttt tagaggcaca 540

aactcgttca gaagcgccat taccgacatc gtatttaatt ttagcgacta taagcccgtt 600

aagggtgcga aagtacatgc cggctttctt agttcatatg aacaggtggt gaatgactac 660

ttcccggtgg tgcaagaaca gctcactgcg aatcctacat acaaagtcat cgttactgga 720

cacagcttag ggggtgcaca agcccttctc gccggaatgg acctatatca acgggagcct 780

cgactctccc caaagaatct tagtatattc actgtaggcg gcccccgcgt aggcaacccc 840

acctttgctt attacgtgga atctactggt attcctttcc aacggacggt tcataagcgc 900

gacatagtac cacacgttcc gccccagtcc ttcgggttct tgcatccagg cgtggagtca 960

tggattaaat cgggtacgtc aaatgtccaa atctgtacat cagaaattga aacaaaagat 1020

tgttcaaact caatagttcc gtttacatcg attctggatc acttgtcata ctttgacatt 1080

aacgaaggtt cgtgcttggg cggcggagga tctggtggcg gcgggtccgg cggaggtggg 1140

agccatgaag aggacggtgt ctgcaactca aatgcgccat gttatcactg tgatgcaaac 1200

ggggaaaatt gctcctgtaa ctgcgagcta ttcgattgcg aagcgaaaaa acccgacggt 1260

tcttatgccc acccatgccg tcgatgtgat gcgaataata tatgtaagtg ttcctgcacg 1320

gctatacctt gtaatgagga ccatccatgt caccactgtc atgaggaaga tgatggtgac 1380

acccactgtc actgtagttg tgagcactcc cacgatcatc acgatgacga tactcatggg 1440

gagtgcacta agaaggcacc gtgttggagg tgtgaataca atgccgatct caagcatgat 1500

gtctgcggat gcgagtgcag taaactacct tgtaacgacg agcatccctg ctatcgtaaa 1560

gaaggaggag tggtgtcttg cgactgcaag acgataacct gcaacgaaga tcatccgtgt 1620

tatcacagct acgaagaaga tggagtaacc aaatcggatt gcgattgcga gcactcccct 1680

ggtccttctg aacatcacca tcatcatcat tga 1713

Claims (7)

1. An expression vector comprising any one of:

i) The vector pAO alpha N-4Mrcp19k has a sequence shown in SEQ ID NO. 1;

ii) pAO alpha N-4Mrcp20k vector, the sequence of which is shown in SEQ ID NO. 2;

iii) The vector pAO alpha N-4proROL-Mrcp19k has a sequence shown in SEQ ID NO. 3;

iv) pAO alpha N-4proROL-Mrcp20k vector, the sequence is shown in SEQ ID NO. 4.

2. A vector combination comprising the vector of claim 1 in combination with a pPICZ-Ssa4-VHb and pPICZ 3.5k-Sso2-Bmh vector;

wherein the sequence of pPICZ-Ssa4-VHb is shown in SEQ ID NO. 5; the sequence of the pPIC3.5k-Sso2-Bmh vector is shown in SEQ ID NO. 6.

3. An engineered yeast strain comprising the expression vector of claim 1 or the combination of vectors of claim 2.

4. The engineered yeast strain of claim 3, wherein the yeast strain is Pichia pastoris GS115.

5. The engineered yeast strain of any one of claims 3 to 4, which is deposited with the China center for type culture Collection with the deposit number CCTCC M2021265 or CCTCC M2021266.

6. The method for expressing mucin by using the engineered yeast strain of any one of claims 3 to 5, wherein the BMGY medium is initially pH =7.0, the inoculum size is 3% (v/v), the inducer methanol is added in an amount of 1.5% (v/v), the induction time is 96h, and the induction temperature is 25 ℃ in the bacterial culture.

7. The method of claim 6, comprising the steps of:

(1) Transforming pichia competent cells by the expression vector or the vector combination of claim 1 or 2 to obtain a yeast recombinant genetic engineering strain containing the expression vector or the vector combination;

(2) Inoculating the yeast recombinant genetic engineering bacteria obtained in the step (1) into 5mL YPD liquid culture medium, and culturing for 20h;

(3) Inoculating the engineering bacteria in a BMGY culture medium with initial pH =7.0 of 50mL at the temperature of 28 ℃ for 24h at the inoculation amount of 3% (v/v);

(4) The thalli are collected by centrifugation and inoculated in 50mL BMMY culture medium, inducer absolute methanol is added every 24h until the final volume concentration of the methanol is 1.5% (v/v), and the mixture is subjected to induced expression for 96h at 25 ℃ on a shaker at 200 rpm/min.

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