CN109722404B - Engineering strain and method for preparing farnesene by using cellulose - Google Patents

Abstract

In order to achieve the above objects, in one aspect, the present disclosure provides an engineered strain of escherichia coli having a accession number of CGMCC No. 14818. The present disclosure also provides a method for preparing engineering bacteria, comprising: transferring the recombinant plasmid into the engineering bacterium host bacterium to enable the transferred engineering bacterium host bacterium to express the gene inserted into the recombinant plasmid; the recombinant plasmid comprises a plasmid shown as SEQ ID NO.1, a plasmid shown as SEQ ID NO.2 and a plasmid shown as SEQ ID NO. 3. The disclosure also provides the engineering bacteria prepared by the method, the engineering strains, the application of the engineering bacteria and a method for preparing farnesene by using cellulose. According to the technical scheme, the yield of farnesene produced by fermentation of the cellulose hydrolysate is obviously improved.

Description

Engineering strain and method for preparing farnesene by using cellulose

Technical Field

The disclosure relates to the field of bioengineering, and in particular relates to an engineering strain, a method for preparing the engineering strain, the engineering strain prepared by the method, application of the engineering strain and the engineering strain, and a method for preparing farnesene by using cellulose.

Background

Farnesene can be hydrogenated and converted into farnesane, and is a renewable fuel which can be mixed with petroleum diesel and aerospace fuel for use. Farnesene has the advantages of no sulfur, high cetane number, low temperature performance and low GHG emission, and is a bio-based chemical with wide application and ideal effect. For example, CN101553558A describes the production of farnesene by the MEV pathway in e.coli using a glucose-containing medium.

One important problem to be solved in the microbial production of farnesene is the source of glucose. Cellulose is a polysaccharide which is widely distributed and has the largest content in the nature, accounts for more than 50 percent of the carbon content in the plant, and is an important glucose source. Specifically, cellulose is a macromolecular polysaccharide formed by connecting D-glucose through beta-1, 4-glycosidic bonds, is insoluble in water and common organic solvents, and is a main component of plant cell walls. For example, the cellulose content of cotton is close to 100%. Cellulose is usually combined with hemicellulose, pectin and lignin. For example, in general wood, cellulose accounts for 40-50%, hemicellulose accounts for 10-30%, and lignin accounts for 20-30%.

Cellulose can be used as a carbon source by microorganism engineering bacteria only after being degraded into glucose. In actual production, raw materials as sources of cellulose mainly include straw, bark, wood chips, corncobs, rice bran, wheat bran, and the like, and cellulose in these raw materials tends to form a supramolecular stable structure having a high crystalline region, and hemicellulose and lignin are closely embedded around the supramolecular stable structure, thereby forming a complex stable structure, which makes it difficult for cellulose to be directly utilized by microorganisms. At present, the raw material as the source of cellulose is usually pretreated and enzymatically hydrolyzed, and the resulting monosaccharide-containing product is fed to the fermentation process. For example, the conventional pretreatment methods mainly include dilute acid treatment, hydrothermal treatment, steam explosion, organic solvent treatment, low-temperature ammonia leaching treatment, alkali treatment, alkaline hydrogen peroxide treatment, biological treatment, and the like. As another example, the enzymatic hydrolysis may include enzymatic hydrolysis with cellulase and/or beta-glucosidase.

However, in the course of the research, it was found that hydrolysis of cellulose to produce a product containing monosaccharides has the disadvantage of low yield if used in the fermentative production of farnesene.

Disclosure of Invention

One of the objectives of the present disclosure is to increase the yield of farnesene produced by fermentation using cellulose hydrolysate.

In order to achieve the above objects, in one aspect, the present disclosure provides an engineered strain of escherichia coli having a accession number of CGMCC No. 14818.

In another aspect, the present disclosure also provides a method for preparing engineering bacteria, comprising: transferring the recombinant plasmid into the engineering bacterium host bacterium to enable the transferred engineering bacterium host bacterium to express the gene inserted into the recombinant plasmid; the recombinant plasmid comprises a pMevT plasmid shown as SEQ ID NO.1, a pMBIS plasmid shown as SEQ ID NO.2 and a pFII plasmid shown as SEQ ID NO. 3.

On the other hand, the disclosure also provides the engineering bacteria prepared by the method.

On the other hand, the disclosure also provides the application of the engineering strain or the engineering bacteria prepared by the method in fermentation of a cellulose hydrolysate to generate farnesene.

In still another aspect, the present disclosure also provides a method for preparing farnesene from cellulose, wherein the method comprises the following steps: (1) hydrolyzing a solid raw material containing cellulose to obtain a hydrolysate containing monosaccharide; (2) inoculating the engineering strain or engineering bacterium into the liquid fermentation raw material containing the hydrolysate containing monosaccharide, and performing fermentation culture to obtain a cultured material.

The yield of farnesene produced by fermentation of cellulose hydrolysate is probably caused by poor stress resistance or poor synthesis capability of the strain, and the stress resistance of the engineering bacteria can be remarkably improved by selecting a proper carrier, a metabolic pathway, an overexpression key metabolic enzyme, a gene arrangement sequence and a promoter for combined application to balance the metabolic pathway. Therefore, the present disclosure significantly improves the yield of farnesene produced by fermentation using cellulose hydrolysate according to the above technical solution.

Additional features and advantages of the disclosure will be set forth in the detailed description which follows.

Biological preservation

The biological preservation information to which the present disclosure relates includes: the classification is named as: escherichia coli; the preservation unit is as follows: china general microbiological culture Collection center; the address is as follows: xilu No.1 Hospital No.3, Beijing, Chaoyang, North; the preservation date is as follows: year 2017, month 10, day 16; the preservation number is: CGMCC NO. 14818.

Detailed Description

The following describes in detail specific embodiments of the present disclosure. It should be understood that the detailed description and specific examples, while indicating the present disclosure, are given by way of illustration and explanation only, not limitation.

In one aspect, the present disclosure provides an engineered strain, wherein the engineered strain is escherichia coli with a collection number of CGMCC No. 14818.

Wherein, the Escherichia coli with the preservation number of CGMCC NO.14818 is a clone with higher activity which is unexpectedly obtained in the process of preparing the engineering bacteria according to the method for preparing the engineering bacteria, so that the engineering strain with better effect is obtained.

The present disclosure also provides a method for preparing engineering bacteria, comprising: transferring the recombinant plasmid into the engineering bacterium host bacterium to enable the transferred engineering bacterium host bacterium to express the gene inserted into the recombinant plasmid; the recombinant plasmid comprises a pMevT plasmid shown as SEQ ID NO.1, a pMBIS plasmid shown as SEQ ID NO.2 and a pFII plasmid shown as SEQ ID NO. 3.

Optionally, wherein the engineering bacterium is escherichia coli.

Optionally, wherein the escherichia coli is escherichia coli DH5 a and/or escherichia coli BL 21.

Wherein, the recombinant plasmid composed of three plasmids, i.e. pMevT plasmid shown as SEQ ID NO.1, pMBIS plasmid shown as SEQ ID NO.2 and pFII plasmid shown as SEQ ID NO.3, enables the engineering bacteria to have the expression of isopentenyl pyrophosphate isomerase (IDI), farnesene pyrophosphate synthetase (ispA) and farnesene synthetase (FG), and can have the expression of various enzymes forming a Mevalonate (MVA) pathway and can also have the over-expression of citrate lyase (ACL); and the recombinant plasmid with the specific combination can enable the expression amount of various metabolic enzymes in the engineering bacteria to form an efficient balanced metabolic pathway, thereby improving the yield of farnesene produced by fermentation of a cellulose hydrolysate.

Wherein the plurality of enzymes comprising the Mevalonate (MVA) pathway comprise acetoacetyl CoA thiolase, HMG-CoA synthase, HMG-CoA reductase, mevalonate kinase (E12), phosphomevalonate kinase (E8), and mevalonate pyrophosphate decarboxylase (MVD 1); the acetoacetyl-CoA thiolase is an atoB enzyme; the HMG-CoA synthase is HMGS enzyme, and the HMG-CoA reductase is HMGR enzyme.

The species source, amino acid sequence, and nucleotide coding sequence of the various enzymes involved in this disclosure may be a known choice, among others. For example, isopentenyl pyrophosphate isomerase (IDI) refers to the enzyme numbered EC 5.3.3.2, the source and sequence of which can be of known species (including artificial sequences). Reference information for the various enzymes involved in the present disclosure is given in table 1, but the selection and use of the various enzymes involved in the present disclosure is not limited to the information in table 1.

TABLE 1

Name of enzyme	Numbering in enzymology	Reference coding sequence
			Isopentenyl pyrophosphate isomerase (IDI)	EC 5.3.3.2	Position 7197-744 in SEQ ID NO.2
Farnesene pyrophosphate synthetase (ispA)	EC 2.5.1.10	Position 7767-8666 of SEQ ID NO.2
			Farnesene synthetase (FG)	EC 4.2.3.47	5069-6792 position of SEQ ID NO.6
atoB enzyme	EC 2.3.1.9	1320-2503 of SEQ ID NO.5
			HMGS enzyme	EC 2.3.3.10	2519-3993 in SEQ ID NO.5
HMGR enzymes	EC 1.1.1.34	4009-7145 in SEQ ID NO.5
			Mevalonate kinase (E12)	EC 2.7.1.36	3535-4565 position in SEQ ID NO.2
Phosphomevalonate kinase (E8)	EC 2.7.4.2	4588-5943 in SEQ ID NO.2
			Mevalonate pyrophosphate decarboxylase (MVD1)	EC 2.5.1.10	5975-7164 in SEQ ID NO.2
Citric acid lyase (ACL)	EC2.3.3.8	6037-7496 position in SEQ ID NO.1

On the other hand, the disclosure also provides the engineering bacteria prepared by the method.

In another aspect, the present disclosure also provides a use of the above engineered strain or the above engineered bacterium in fermentation of a cellulose hydrolysate to produce farnesene.

In another aspect, the present disclosure also provides a method for preparing farnesene from cellulose, wherein the method comprises the following steps: (1) hydrolyzing a solid raw material containing cellulose to obtain a hydrolysate containing monosaccharide; (2) inoculating the engineering strain or the engineering bacteria into the liquid fermentation raw material containing the hydrolysate containing the monosaccharide, and performing fermentation culture to obtain a cultured material.

Optionally, wherein the cellulose-containing material comprises at least one of straw, bark, wood chips, corn cobs, rice bran, and wheat bran.

Optionally, wherein the step of subjecting the raw material containing cellulose to hydrolysis treatment comprises: s1, sequentially carrying out acid pretreatment and alkali pretreatment on the solid raw material containing cellulose, or sequentially carrying out alkali pretreatment and acid pretreatment to obtain a pretreated solid material; s2, carrying out enzymatic hydrolysis on the pretreated solid material.

Optionally, wherein the acid pretreatment is formic acid treatment and/or dilute sulfuric acid treatment; the formic acid treatment conditions include: the temperature is 40-100 ℃, the time is 1-10 hours, the volume of a formic acid solution is 5-50 liters per kilogram of pretreated solid material, and the formic acid solution contains 70-95 wt% of formic acid and 0-2 wt% of HCl; the conditions of the dilute sulfuric acid treatment include: the temperature is 80-150 ℃, the time is 5 minutes-2 hours, and the volume of the dilute sulfuric acid solution is 5-50 liters per kilogram of pretreated solid material, wherein the dilute sulfuric acid solution contains 0.5-10 wt% of dilute sulfuric acid.

Optionally, wherein the alkali pretreatment is ammonia treatment and/or sodium hydroxide treatment; the ammonia water treatment conditions comprise: the temperature is 40-80 ℃, the time is 2-24 hours, the volume of ammonia water is 2-20 liters per kilogram of pretreated solid material, and the ammonia water contains 10-25 wt% of NH₃(ii) a The sodium hydroxide treatment conditions include: the temperature is 60-120 ℃, the time is 1-12 hours, and the volume of the sodium hydroxide aqueous solution is 5-50 liters per kilogram of pretreated solid material, wherein the sodium hydroxide aqueous solution contains 0.5-10 wt% of sodium hydroxide.

Optionally, wherein the conditions of the enzymatic hydrolysis comprise: the temperature is 30-70 ℃, the time is 3-72 hours, the pH is 4.0-6.0, the volume of a buffer solution is 5-100 liters per kilogram of pretreated solid materials, the buffer solution used for enzymatic hydrolysis contains 0.01-1 mol/L citric acid buffer solution and/or 0.01-1 mol/L acetic acid buffer solution, the enzyme used for enzymatic hydrolysis is cellulase and/or beta-glucosidase, the enzyme adding amount of the cellulase is 10-500U per gram of pretreated solid materials, and the enzyme adding amount of the beta-glucosidase is 10-250U per gram of pretreated solid materials.

Optionally, wherein the conditions of the fermentation culture comprise: the temperature is 16-35 ℃, the time is 3-120 h, the dosage of the extracting agent is 50-1000 mL relative to each liter of the liquid fermentation raw material, and the inoculation amount of the engineering bacteria or the engineering strains is (10-300) multiplied by 10⁷A plurality of; the extractant is selected from n-hexane, n-decane, dodecane or kerosene; and in each liter of the liquid fermentation raw material, the content of the hydrolysate containing monosaccharide is 0.01-0.9 kg calculated by the amount of monosaccharide.

The invention is illustrated in further detail by the following examples:

example 1

This example illustrates the preparation of a cellulase hydrolysate.

100g of corn straw with the particle size of 20-80 meshes is weighed, 1.5L of formic acid solution (containing 88 wt% of formic acid and 1 wt% of HCl) is added, and the mixture is placed in a treatment tank. Reacting in a constant temperature water bath at 65 ℃ for 3h, and then carrying out solid-liquid separation on the reaction slurry. And (3) carrying out ammonia water treatment on the solid component, wherein the treatment conditions are as follows: the ammonia concentration was 15% by weight, the volume of ammonia used per kg of solid component was 8 liters, the treatment temperature was 60 ℃ and the reaction time was 16 hours. And after the reaction is finished, carrying out solid-liquid separation on the material, washing the solid component to pH 6-7 with distilled water, and drying to obtain the pretreated solid material.

Carrying out enzymatic hydrolysis on the pretreated solid material, wherein the enzymatic hydrolysis conditions comprise: the temperature is 50 ℃, the volume of a buffer solution used by each kilogram of pretreated solid materials is 20 liters, the buffer solution is a citric acid buffer solution (0.15mol/L) with the pH value of 4.8, enzymes used for enzymatic hydrolysis are cellulase and beta-glucosidase, the enzyme adding amount of the cellulase is 30U per gram of pretreated solid materials, and the enzyme adding amount of the beta-glucosidase is 60U per gram of pretreated solid materials. Taking out the reaction slurry after enzymolysis for 24h, inactivating the enzyme in a boiling water bath for 10min, and then carrying out suction filtration. And analyzing and detecting the liquid component obtained by suction filtration by using HPLC (high performance liquid chromatography), wherein the glucose concentration of the obtained cellulose enzymolysis liquid is 38.2 g/L.

Example 2

This example serves to illustrate the sequence structure of the plasmid. Commercially available vectors used in this example include the pBAD33 vector, the pACYDCDuet-1 vector, the pBBR1MCS-4 vector, and the pet-28a vector.

According to the instruction of the vector, molecular biological operations such as PCR cloning, whole gene synthesis, restriction enzyme digestion and connection are adopted to construct 1 set of MVA metabolic path plasmid, specifically including pMevT plasmid, pMBIS plasmid and pFII plasmid, 1 plasmid for replacing pFII plasmid, namely pFG plasmid, and then 2 plasmids for replacing pMevT plasmid, namely pMevT-1 and pMevT-2, are constructed. The sequence information of the above plasmids is specifically shown in Table 2.

TABLE 2

Example 3

This example illustrates the construction of engineered strains.

Construction of experimental group engineered strain F1: selecting Escherichia coli BL21 as host cell, passing through CaCl₂Transformation methods were transferred to the pMevT, pMBIS and pFII plasmids described in table 2. The strain can synthesize farnesene through an MVA metabolic pathway, has an optimized high-efficiency balanced metabolic pathway, and has overexpression of citrate lyase (ACL). On an antibiotic resistance screening plate, 9 clones of the strain are selected, namely F1-1, F1-2, F1-3, F1-4, F1-5, F1-6, F1-7, F1-8 and F1-9.

Construction of control engineered strain DF 1: selecting Escherichia coli BL21 as host cell, passing through CaCl₂Transformation methods were transferred to the pMevT-1 plasmid, pMBIS plasmid and pFG plasmid described in Table 2. The strain can synthesize farnesene through MVA metabolic pathway, but is not balancedOptimization of metabolic pathways. On the antibiotic resistance selection plate, 3 clones of the strain were selected, DF1-1, DF1-2 and DF1-3, respectively.

Construction of control engineered strain DF 2: selecting Escherichia coli BL21 as host cell, passing through CaCl₂Transformation methods were transferred to the pMevT-2 plasmid, pMBIS plasmid and pFG plasmid described in Table 2. The strain can synthesize farnesene through an MVA metabolic pathway, has an optimized efficient balanced metabolic pathway, and does not have the over-expression of citrate lyase (ACL). On the antibiotic resistance selection plate, 3 clones of the strain were selected, DF2-1, DF2-2 and DF2-3, respectively.

Example 4

This example illustrates the preparation of farnesene from cellulose.

The cellulase hydrolysate obtained in example 1 was used as a hydrolysate containing monosaccharides to prepare a liquid fermentation raw material. The liquid fermentation raw material contained 100mL of the cellulase hydrolysate obtained in example 1 per liter, 10g of peptone, 5g of yeast extract, 10g of NaCl, and the balance of water. And sterilizing the liquid fermentation raw material for later use.

Antibiotics (chloramphenicol, kanamycin and ampicillin at a final concentration of 30mg/mL) were added to the above liquid fermentation raw materials, and then 15 clones obtained in example 3 were inoculated, respectively, at an inoculation density of 200X 10⁷And (2) per liter. Cultured to OD at 37 ℃₆₀₀Adding IPTG (isopropyl thiogalactoside) with the final concentration of 200mg/L, L-arabinose with the final concentration of 1g/L and 20mL of n-decane into the fermentation liquor between 0.6 and 08 ℃, and performing induction expression at 30 ℃ to synthesize farnesene by fermentation. After 24h of induction, 1mL of the fermentation broth and 100. mu.L of the extraction layer (n-decane) were used to determine the cell density and farnesene concentration, respectively. The results are shown in Table 3.

TABLE 3

As can be seen from the data in table 3, 9 clones F1-1, F1-2, F1-3, F1-4, F1-5, F1-6, F1-7, F1-8 and F1-9 of the experimental engineered strain F1 were all significantly higher than 3 clones DF1-1, DF1-2 and DF1-3 of the control engineered strain DF1, and also significantly higher than 3 clones DF2-1, DF2-2 and DF2-3 of the control engineered strain DF2, and of 9 clones of the experimental engineered strain F1, F1-3 had the highest farnesene content in the fermentation product, which was selected as the optimal strain to be deposited with the accession number: CGMCC NO. 14818. It can be seen that the co-transformation of the pMevT, pMBIS and pFII plasmids described in table 2 significantly improved the yield of farnesene produced by fermentation using cellulose hydrolysate, and more preferably, the accession number: CGMCC NO.14818 has more outstanding conversion effect.

The preferred embodiments of the present disclosure have been described in detail above, however, the present disclosure is not limited to the specific details of the above embodiments, and various simple modifications may be made to the technical solution of the present disclosure within the technical idea of the present disclosure, and these simple modifications all fall within the protection scope of the present disclosure.

It should be noted that, in the foregoing embodiments, various features described in the above embodiments may be combined in any suitable manner, and in order to avoid unnecessary repetition, various combinations that are possible in the present disclosure are not described again.

In addition, any combination of various embodiments of the present disclosure may be made, and the same should be considered as the disclosure of the present disclosure, as long as it does not depart from the spirit of the present disclosure.

Sequence listing

<110> China petrochemical Co., Ltd

Research Institute of petrochemicals, Sinopec

<120> engineering strain and method for preparing farnesene by using cellulose

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agaggcccag tcgttcgttt ccctacttta ataagatctg gtgcctgcaa gatatggtta 5040

gactcggaag agggacaaaa ttcaattaaa aaagctttta attctacatc aaggtttgca 5100

cgtttgcaac atattcaaac ctgtctagca ggcgatttgc tttttatgag atttcggaca 5160

actaccggtg acgcaatggg tatgaacatg atatcgaaag gtgtcgaata ctctttgaaa 5220

caaatggtag aagaatatgg ttgggaagat atggaagttg tctccgtatc tggtaactat 5280

tgtactgata agaaacctgc cgcaatcaat tggattgaag gtcgtggtaa aagtgtcgta 5340

gctgaagcta ctattcctgg tgatgtcgta aaaagtgttt taaagagcga tgtttccgct 5400

ttagttgaat taaatatatc caagaacttg gttggatccg caatggctgg atctgttggt 5460

ggtttcaacg cgcacgcagc taatttggtc actgcacttt tcttggcatt aggccaagat 5520

cctgcgcaga acgtcgaaag ttccaactgt ataactttga tgaaggaagt tgatggtgat 5580

ttaaggatct ctgtttccat gccatctatt gaagttggta cgattggcgg gggtactgtt 5640

ctggagcctc agggcgccat gcttgatctt ctcggcgttc gtggtcctca ccccactgaa 5700

cctggagcaa atgctaggca attagctaga ataatcgcgt gtgctgtctt ggctggtgaa 5760

ctgtctctgt gctccgcact tgctgccggt cacctggtac aaagccatat gactcacaac 5820

cgtaaaacaa acaaagccaa tgaactgcca caaccaagta acaaagggcc cccctgtaaa 5880

acctcagcat tattataatg cttaagtcga acagaaagta atcgtattgt acacggccgc 5940

ataatcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attccccatc 6000

ttagtatatt agttaagtat aaaggaggaa ttaaccatgt ctgccaagtc aatccttgaa 6060

gcggatggca aggccattct caactaccac ctcaccagaa gccccgtcat caaatcctcc 6120

cctctacccc ccccatccac ccacaatccc cccccaaagc tcgcctccct ctacttcccc 6180

gaagatgcct ctgttgacgc catcctcgac caggctgaga gcacctatcc ctggctgctc 6240

gcaaaggata tccgcctcgt cgcaaaacca gaccaattga tcaagcggcg gggcaagagt 6300

ggcctgcttg ctctggacag gacctggcct gaagcgagag catggattgc tgagcgcgca 6360

ggcaaggagc agaaggttga gactgttacg ggtgtcctga ggcagttcct cgttgaacca 6420

tttgtgcctc acccgcaaga aacggaatat tatatcaaca tcaactccgt acgagagggc 6480

gactggatcc tcttcaccca cgaaggcggc gtagatgtcg gcgacgtcga cgccaaggca 6540

gaaaagctct tgatcccagt caatctcgca aactacccct ccaacgacga aatcgcctct 6600

gccctcctca gcaaaatccc ctcgggcctc cacaacgtgc tcgtcgactt catcacccgc 6660

ctctacgccg tctacgtcga ctgccaattc acctatctcg aaatcaaccc cctcgttgtc 6720

attcccaatc cctccaaaac ctccgcagca gtgcacttcc ttgacctagc cgccaaacta 6780

gaccaaactg ccgaattcga atgcggtacc aaatgggcca ttgcccgcag ccccgccgct 6840

ctggggatcc gcaaggccac caatacggat ggcaaagtaa ccatcgacgc ggggccgccc 6900

atggaattcc ccgccccctt cggccgcgag cttagcaagg aagaaaaatt catcgcagac 6960

atggacgcca agacaggctc gtcgcttaaa ctcaccgtcc tcaacccgaa cggacgcatc 7020

tggaccctcg tcgccggcgg tggtgcatcc gtcgtctacg ccgacgccat cgcgtccgct 7080

ggcttcgtct ccgaactcgc aaactacgga gaatactccg gcgcacccac cgaaacgcag 7140

acattcaact acgcacgcac catcctggat ctgatgctgc gcgcaccctt gcatccggat 7200

gggaaggtcc tgttcattgg tggtgggatt gcgaatttca cgaacgtcgc atcgacgttc 7260

aagggggtta ttcgggcgct gagggaggtg gcgcctatct tgaatgagca taagacacag 7320

atctgggtta ggcgggctgg gcctaattac caggaaggcc tgaagaatat taaggctgtg 7380

ggcgaggagt tggggttgga tatgcatgtt tatgggccag agatgcatgt tagtgggatc 7440

gtgccgttgg cattgttggg gaagaagacc gatgttaagg aatttggagc ggcgtagtta 7500

acctaggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg 7560

gtcttgaggg gttttttgct gaaacctcag gcatttgaga agcacacggt cacactgctt 7620

ccggtagtca ataaaccggt aaaccagcaa tagacataag cggctattta acgaccctgc 7680

cctgaaccga cgaccgggtc gaatttgctt tcgaatttct gccattcatc cgcttattat 7740

cacttattca ggcgtagcac caggcgttta agggcaccaa taactgcctt aaaaaaatta 7800

cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg 7860

gaagccatca cagacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc 7920

ttgcgtataa tatttgccca tagtgaaaac gggggcgaag aagttgtcca tattggccac 7980

gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc 8040

aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata 8100

tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc 8160

agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc 8220

gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat 8280

aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc 8340

cagctgaacg gtctggttat aggtacattg agcaactgac tgaaatgcct caaaatgttc 8400

tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt 8460

agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat 8520

ttcattatgg tgaaagttgg aacctcttac gtgccgatca acgtctcatt ttcgccaaaa 8580

gttggcccag ggcttcccgg tatcaacagg gacaccagga tttatttatt ctgcgaagtg 8640

atcttccgtc acaggtattt attcggcgca aagtgcgtcg ggtgatgctg ccaacttact 8700

gatttagtgt atgatggtgt ttttgaggtg ctccagtggc ttctgtttct atcagctgtc 8760

cctcctgttc agctactgac ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc 8820

gctagcggag tgtatactgg cttactatgt tggcactgat gagggtgtca gtgaagtgct 8880

tcatgtggca ggagaaaaaa ggctgcaccg gtgcgtcagc agaatatgtg atacaggata 8940

tattccgctt cctcgctcac tgactcgcta cgctcggtcg ttcgactgcg gcgagcggaa 9000

atggcttacg aacggggcgg agatttcctg gaagatgcca ggaagatact taacagggaa 9060

gtgagagggc cgcggcaaag ccgtttttcc ataggctccg cccccctgac aagcatcacg 9120

aaatctgacg ctcaaatcag tggtggcgaa acccgacagg actataaaga taccaggcgt 9180

ttcccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat 9240

tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag 9300

ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat 9360

ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag 9420

ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact 9480

gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg 9540

gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa 9600

gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat 9660

ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga 9720

tataagttgt aattctcatg ttagtcatgc cccgcgccca ccggaaggag ctgactgggt 9780

tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc taacttacat 9840

taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt 9900

aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag ggtggttttt 9960

cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc ctgagagagt 10020

tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt 10080

aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac cgagatgtcc 10140

gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg 10200

ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat ggtttgttga 10260

aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat ttgattgcga 10320

gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact taatgggccc 10380

gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc cagtcgcgta 10440

ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac atcaagaaat 10500

aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc atccagcgga 10560

tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag 10620

gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag ttgatcggcg 10680

cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact ggaggtggca 10740

acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa 10800

ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac gtggctggcc 10860

tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc gacatcgtat 10920

aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg ctatcatgcc 10980

ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct ctcccttatg 11040

cgactcctgc attaggaaat taatacgact cactata 11077

<210> 2

<211> 10396

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 2

ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg 60

cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt 120

ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg 180

cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg 240

cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca 300

gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt 360

tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg 420

tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt 480

acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg 540

ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct 600

tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg 660

ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc 720

ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca 780

tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg 840

cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg 900

cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc 960

ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc 1020

gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga 1080

tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc 1140

ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt 1200

ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg 1260

acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac 1320

gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc 1380

tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg 1440

caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg 1500

gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag 1560

aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta 1620

cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg 1680

taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc 1740

tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac 1800

gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga 1860

acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct 1920

gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac 1980

actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt 2040

ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt 2100

ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag 2160

tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct 2220

gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga 2280

gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac 2340

ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct 2400

ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt 2460

gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg 2520

ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga 2580

cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga 2640

ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg 2700

gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa 2760

ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa 2820

tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag 2880

gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc 2940

agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca 3000

aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg 3060

atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg 3120

attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga 3180

gcgcgcgtaa tacgactcac tatagggcga attggagctc aggaggaatt aaccatgtca 3240

ttaccgttct taacttctgc accgggaaag gttattattt ttggtgaaca ctctgctgtg 3300

tacaacaagc ctgccgtcgc tgctagtgtg tctgcgttga gaacctacct gctaataagc 3360

gagtcatctg caccagatac tattgaattg gacttcccgg acattagctt taatcataag 3420

tggtccatca atgatttcaa tgccatcacc gaggatcaag taaactccca aaaattggcc 3480

aaggctcaac aagccaccga tggcttgtct caggaactcg ttagtctttt ggatccgttg 3540

ttagctcaac tatccgaatc cttccactac catgcagcgt tttgtttcct gtatatgttt 3600

gtttgcctat gcccccatgc caagaatatt aagttttctt taaagtctac tttacccatc 3660

ggtgctgggt tgggctcaag cgcctctatt tctgtatcac tggccttagc tatggcctac 3720

ttgggggggt taataggatc taatgacttg gaaaagctgt cagaaaacga taagcatata 3780

gtgaatcaat gggccttcat aggtgaaaag tgtattcacg gtaccccttc aggaatagat 3840

aacgctgtgg ccacttatgg taatgccctg ctatttgaaa aagactcaca taatggaaca 3900

ataaacacaa acaattttaa gttcttagat gatttcccag ccattccaat gatcctaacc 3960

tatactagaa ttccaaggtc tacaaaagat cttgttgctc gcgttcgtgt gttggtcacc 4020

gagaaatttc ctgaagttat gaagccaatt ctagatgcca tgggtgaatg tgccctacaa 4080

ggcttagaga tcatgactaa gttaagtaaa tgtaaaggca ccgatgacga ggctgtagaa 4140

actaataatg aactgtatga acaactattg gaattgataa gaataaatca tggactgctt 4200

gtctcaatcg gtgtttctca tcctggatta gaacttatta aaaatctgag cgatgatttg 4260

agaattggct ccacaaaact taccggtgct ggtggcggcg gttgctcttt gactttgtta 4320

cgaagagaca ttactcaaga gcaaattgac agcttcaaaa agaaattgca agatgatttt 4380

agttacgaga catttgaaac agacttgggt gggactggct gctgtttgtt aagcgcaaaa 4440

aatttgaata aagatcttaa aatcaaatcc ctagtattcc aattatttga aaataaaact 4500

accacaaagc aacaaattga cgatctatta ttgccaggaa acacgaattt accatggact 4560

tcataaactt cataggaggc agatcaaatg tcagagttga gagccttcag tgccccaggg 4620

aaagcgttac tagctggtgg atatttagtt ttagatacaa aatatgaagc atttgtagtc 4680

ggattatcgg caagaatgca tgctgtagcc catccttacg gttcattgca agggtctgat 4740

aagtttgaag tgcgtgtgaa aagtaaacaa tttaaagatg gggagtggct gtaccatata 4800

agtcctaaaa gtggcttcat tcctgtttcg ataggcggat ctaagaaccc tttcattgaa 4860

aaagttatcg ctaacgtatt tagctacttt aaacctaaca tggacgacta ctgcaataga 4920

aacttgttcg ttattgatat tttctctgat gatgcctacc attctcagga ggatagcgtt 4980

accgagcatc gtggcaacag aagattgagt tttcattcgc acagaattga agaagttccc 5040

aaaacagggc tgggctcctc ggcaggttta gtcacagttt taactacagc tttggcctcc 5100

ttttttgtat cggacctgga aaataatgta gacaaatata gagaagttat tcataattta 5160

gcacaagttg ctcattgtca agctcagggt aaaattggaa gcgggtttga tgtagcggcg 5220

gcagcatatg gatctatcag atatagaaga ttcccacccg cattaatctc taatttgcca 5280

gatattggaa gtgctactta cggcagtaaa ctggcgcatt tggttgatga agaagactgg 5340

aatattacga ttaaaagtaa ccatttacct tcgggattaa ctttatggat gggcgatatt 5400

aagaatggtt cagaaacagt aaaactggtc cagaaggtaa aaaattggta tgattcgcat 5460

atgccagaaa gcttgaaaat atatacagaa ctcgatcatg caaattctag atttatggat 5520

ggactatcta aactagatcg cttacacgag actcatgacg attacagcga tcagatattt 5580

gagtctcttg agaggaatga ctgtacctgt caaaagtatc ctgaaatcac agaagttaga 5640

gatgcagttg ccacaattag acgttccttt agaaaaataa ctaaagaatc tggtgccgat 5700

atcgaacctc ccgtacaaac tagcttattg gatgattgcc agaccttaaa aggagttctt 5760

acttgcttaa tacctggtgc tggtggttat gacgccattg cagtgattac taagcaagat 5820

gttgatctta gggctcaaac cgctaatgac aaaagatttt ctaaggttca atggctggat 5880

gtaactcagg ctgactgggg tgttaggaaa gaaaaagatc cggaaactta tcttgataaa 5940

taagcggccg cacgataaat aggaggtaat actcatgacc gtttacacag catccgttac 6000

cgcacccgtc aacatcgcaa cccttaagta ttgggggaaa agggacacga agttgaatct 6060

gcccaccaat tcgtccatat cagtgacttt atcgcaagat gacctcagaa cgttgacctc 6120

tgcggctact gcacctgagt ttgaacgcga cactttgtgg ttaaatggag aaccacacag 6180

catcgacaat gaaagaactc aaaattgtct gcgcgaccta cgccaattaa gaaaggaaat 6240

ggaatcgaag gacgcctcat tgcccacatt atctcaatgg aaactccaca ttgtctccga 6300

aaataacttt cctacagcag ctggtttagc ttcctccgct gctggctttg ctgcattggt 6360

ctctgcaatt gctaagttat accaattacc acagtcaact tcagaaatat ctagaatagc 6420

aagaaagggg tctggttcag cttgtagatc gttgtttggc ggatacgtgg cctgggaaat 6480

gggaaaagct gaagatggtc atgattccat ggcagtacaa atcgcagaca gctctgactg 6540

gcctcagatg aaagcttgtg tcctagttgt cagcgatatt aaaaaggatg tgagttccac 6600

tcagggtatg caattgaccg tggcaacctc cgaactattt aaagaaagaa ttgaacatgt 6660

cgtaccaaag agatttgaag tcatgcgtaa agccattgtt gaaaaagatt tcgccacctt 6720

tgcaaaggaa acaatgatgg attccaactc tttccatgcc acatgtttgg actctttccc 6780

tccaatattc tacatgaatg acacttccaa gcgtatcatc agttggtgcc acaccattaa 6840

tcagttttac ggagaaacaa tcgttgcata cacgtttgat gcaggtccaa atgctgtgtt 6900

gtactactta gctgaaaatg agtcgaaact ctttgcattt atctataaat tgtttggctc 6960

tgttcctgga tgggacaaga aatttactac tgagcagctt gaggctttca accatcaatt 7020

tgaatcatct aactttactg cacgtgaatt ggatcttgag ttgcaaaagg atgttgccag 7080

agtgatttta actcaagtcg gttcaggccc acaagaaaca aacgaatctt tgattgacgc 7140

aaagactggt ctaccaaagg aataactgca ggtaatcccg ggaggaggat tactatatgc 7200

aaacggaaca cgtcatttta ttgaatgcac agggagttcc cacgggtacg ctggaaaagt 7260

atgccgcaca cacggcagac acccgcttac atctcgcgtt ctccagttgg ctgtttaatg 7320

ccaaaggaca attattagtt acccgccgcg cactgagcaa aaaagcatgg cctggcgtgt 7380

ggactaactc ggtttgtggg cacccacaac tgggagaaag caacgaagac gcagtgatcc 7440

gccgttgccg ttatgagctt ggcgtggaaa ttacgcctcc tgaatctatc tatcctgact 7500

ttcgctaccg cgccaccgat ccgagtggca ttgtggaaaa tgaagtgtgt ccggtatttg 7560

ccgcacgcac cactagtgcg ttacagatca atgatgatga agtgatggat tatcaatggt 7620

gtgatttagc agatgtatta cacggtattg atgccacgcc gtgggcgttc agtccgtgga 7680

tggtgatgca ggcgacaaat cgcgaagcca gaaaacgatt atctgcattt acccagctta 7740

aataactgca gtaaggagga taccctatgg actttccgca gcaactcgaa gcctgcgtta 7800

agcaggccaa ccaggcgctg agccgtttta tcgccccact gccctttcag aacactcccg 7860

tggtcgaaac catgcagtat ggcgcattat taggtggtaa gcgcctgcga cctttcctgg 7920

tttatgccac cggtcatatg ttcggcgtta gcacaaacac gctggacgca cccgctgccg 7980

ccgttgagtg tatccacgct tactcattaa ttcatgatga tttaccggca atggatgatg 8040

acgatctgcg tcgcggtttg ccaacctgcc atgtgaagtt tggcgaagca aacgcgattc 8100

tcgctggcga cgctttacaa acgctggcgt tctcgatttt aagcgatgcc gatatgccgg 8160

aagtgtcgga ccgcgacaga atttcgatga tttctgaact ggcgagcgcc agtggtattg 8220

ccggaatgtg cggtggtcag gcattagatt tagacgcgga aggcaaacac gtacctctgg 8280

acgcgcttga gcgtattcat cgtcataaaa ccggcgcatt gattcgcgcc gccgttcgcc 8340

ttggtgcatt aagcgccgga gataaaggac gccgtgctct gccggtactc gacaagtatg 8400

cagagagcat cggccttgcc ttccaggttc aggacgacat cctggatgtg gtgggagata 8460

ctgcaacgtt gggaaaacgc cagggtgccg accagcaact tggtaaaagt acctaccctg 8520

cacttctggg tcttgagcaa gcccggaaga aagcccggga tctgatcgac gatgcccgtc 8580

agtcgctgaa acaactggct gaacagtcac tcgatacctc ggcactggaa gcgctagcgg 8640

actacatcat ccagcgtaat aaataacacc gcggtggcgg ccgctctaga actagtggat 8700

cccccgggct gcaggaattc gatatcaagc ttatcgatac cgtcgacctc gagggggggc 8760

ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg 8820

tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc 8880

ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg 8940

ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc 9000

ggccaacgcg cggggagagg cggtttgcgt attgggcgca tgcataaaaa ctgttgtaat 9060

tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac ctgaatcgcc 9120

agcggcatca gcaccttgtc gccttgcgta taatatttgc ccattcaaat atgtatccgc 9180

tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 9240

ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 9300

ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 9360

gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 9420

gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 9480

acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 9540

actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 9600

ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 9660

cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 9720

gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 9780

caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 9840

aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 9900

ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 9960

tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 10020

ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 10080

ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 10140

ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 10200

tccaggtggc acttttcggg gaaatgtgcg cgcccgcgtt cctgctggcg ctgggcctgt 10260

ttctggcgct ggacttcccg ctgttccgtc agcagctttt cgcccacggc cttgatgatc 10320

gcggcggcct tggcctgcat atcccgattc aacggcccca gggcgtccag aacgggcttc 10380

aggcgctccc gaaggt 10396

<210> 3

<211> 8392

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 3

tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60

cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120

ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180

gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240

acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300

ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360

ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420

acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480

tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540

tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600

tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660

actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720

gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780

aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840

agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900

cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960

aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020

tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080

tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140

taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200

ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260

tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320

tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380

cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440

cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500

gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560

gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620

agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680

aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740

agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800

cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860

accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920

aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980

ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040

cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100

gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160

tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220

agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280

tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340

caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400

ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460

gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520

gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580

gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640

aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700

ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760

acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820

ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880

tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940

tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000

cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060

gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120

ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc 3180

catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa 3240

ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc 3300

gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac 3360

gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca 3420

ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta 3480

atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 3540

cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 3600

tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca 3660

ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa 3720

aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt 3780

atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg 3840

cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca 3900

gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta 3960

tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg 4020

agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat 4080

gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct 4140

ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg 4200

catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat 4260

tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc 4320

tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca 4380

gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg 4440

ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt 4500

tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg 4560

catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct 4620

cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga 4680

tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg 4740

ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc 4800

ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg 4860

cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg 4920

gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga 4980

aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa 5040

ttttgtttaa ctttaggagg aattaaccat gagtacactg ccgatcagta gtgtgagctt 5100

cagtagcagc accagcccgc tggtggtgga cgataaagtg agtaccaagc aggacgttat 5160

tcgccatacc atgaacttta acgcaagcat ctggggcgac cagttcctga cctacgatga 5220

accggaggat ctggtgatga agaagcagct ggtggaagaa ttaaaagaag aagttaaaaa 5280

agaactgatt accattaaag gcagcaatga gccgatgcag cacgtgaagc tgctggagct 5340

gattgatgca gttcagcgcc tgggcatcgc ctatcatttt gaagaagaaa tcgaggaagc 5400

actgcagcac atccatgtta cctacggtga acagtgggtg gacaaagaga acctgcaaag 5460

catcagcctg tggttccgct tactgcgcca gcaaggtttc aacgtgagca gcggcgtgtt 5520

caaagatttc atggatgaga aaggtaaatt taaggagagt ctgtgcaatg atgcacaggg 5580

catcttagcc ctgtacgagg ccgcctttat gcgcgtggag gacgaaacca tcctggacaa 5640

cgcactggaa ttcaccaaag tgcatctgga tatcattgcc aaggacccga gttgcgacag 5700

cagcctgcgt acacagattc atcaagcact gaaacaacct ttacgtcgcc gcctggcacg 5760

cattgaagcc ctgcactata tgccgatcta tcagcaggag accagccaca acgaagttct 5820

gttaaagctg gccaagctgg acttcagcgt gctgcagagc atgcacaaga aagaactgag 5880

ccatatttgt aaatggtgga aggacttaga tctgcagaac aaactgccgt atgtgcgtga 5940

tcgcgttgtt gagggttatt tttggattct gagtatctat tacgagccgc agcatgcccg 6000

tacccgcatg ttcctgatga agacatgcat gtggctggtg gtgctggacg acacctttga 6060

taactacggc acctacgagg agctggaaat tttcacccag gccgtggaac gctggagcat 6120

tagctgtctg gatatgctgc cggagtacat gaaattaatt taccaggaac tggtgaacct 6180

gcatgttgag atggaggaga gtctggaaaa agaaggtaag acataccaga tccattacgt 6240

taaagagatg gccaaggagc tggtgcgtaa ttacctggtg gaggcccgtt ggctgaaaga 6300

gggctatatg ccgaccctgg aagagtacat gagcgttagt atggtgaccg gtacctatgg 6360

cctgatgatt gcccgcagct atgttggtcg cggtgatatc gtgacagagg atacctttaa 6420

gtgggtgagc agttacccgc cgatcattaa ggccagctgt gtgatcgtgc gcctgatgga 6480

cgatatcgtg agccataaag aagaacagga gcgtggccat gttgccagca gtatcgagtg 6540

ctacagcaaa gagagtggcg ccagtgaaga agaggcatgt gagtacatca gcggcaaagt 6600

tgaggacgcc tggaaagtga ttaaccgcga aagcctgcgt ccgacagcag ttccgttccc 6660

gctgctgatg ccggccatta atctggcccg tatgtgtgag gttctgtaca gcgtgaacga 6720

tggctttacc catgccgagg gcgacatgaa aagctatatg aaaagctttt ttgtgcaccc 6780

gatggtggtt taaaggagga attaaccatg gactttccgc agcaactcga agcctgcgtt 6840

aagcaggcca accaggcgct gagccgtttt atcgccccac tgccctttca gaacactccc 6900

gtggtcgaaa ccatgcagta tggcgcatta ttaggtggta agcgcctgcg acctttcctg 6960

gtttatgcca ccggtcatat gttcggcgtt agcacaaaca cgctggacgc acccgctgcc 7020

gccgttgagt gtatccacgc ttactcatta attcatgatg atttaccggc aatggatgat 7080

gacgatctgc gtcgcggttt gccaacctgc catgtgaagt ttggcgaagc aaacgcgatt 7140

ctcgctggcg acgctttaca aacgctggcg ttctcgattt taagcgatgc cgatatgccg 7200

gaagtgtcgg accgcgacag aatttcgatg atttctgaac tggcgagcgc cagtggtatt 7260

gccggaatgt gcggtggtca ggcattagat ttagacgcgg aaggcaaaca cgtacctctg 7320

gacgcgcttg agcgtattca tcgtcataaa accggcgcat tgattcgcgc cgccgttcgc 7380

cttggtgcat taagcgccgg agataaagga cgtcgtgctc tgccggtact cgacaagtat 7440

gcagagagca tcggccttgc cttccaggtt caggatgaca tcctggatgt ggtgggagat 7500

actgcaacgt tgggaaaacg ccagggtgcc gaccagcaac ttggtaaaag tacctaccct 7560

gcacttctgg gtcttgagca agcccggaag aaagcccggg atctgatcga cgatgcccgt 7620

cagtcgctga aacaactggc tgaacagtca ctcgatacct cggcactgga agcgctagcg 7680

gactacatca tccagcgtaa taaataaagg aggaattaac catgcaaacg gaacacgtca 7740

ttttattgaa tgcacaggga gttcccacgg gtacgctgga aaagtatgcc gcacacacgg 7800

cagacacccg cttacatctc gcgttctcca gttggctgtt taatgccaaa ggacaattat 7860

tagttacccg ccgcgcactg agcaaaaaag catggcctgg cgtgtggact aactcggttt 7920

gtgggcaccc acaactggga gaaagcaacg aagacgcagt gatccgccgt tgccgttatg 7980

agcttggcgt ggaaattacg cctcctgaat ctatctatcc tgactttcgc taccgcgcca 8040

ccgatccgag tggcattgtg gaaaatgaag tgtgtccggt atttgccgca cgcaccacta 8100

gtgcgttaca gatcaatgat gatgaagtga tggattatca atggtgtgat ttagcagatg 8160

tattacacgg tattgatgcc acgccgtggg cgttcagtcc gtggatggtg atgcaggcga 8220

caaatcgcga agccagaaaa cgattatctg catttaccca gcttaaataa caaagcccga 8280

aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc 8340

tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg at 8392

<210> 4

<211> 9746

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaatagg 60

aggaattaac catgaaaaat tgtgtcatcg tcagtgcggt acgtactgct atcggtagtt 120

ttaacggttc actcgcttcc accagcgcca tcgacctggg ggcgacagta attaaagccg 180

ccattgaacg tgcaaaaatc gattcacaac acgttgatga agtgattatg ggtaacgtgt 240

tacaagccgg gctggggcaa aatccggcgc gtcaggcact gttaaaaagc gggctggcag 300

aaacggtgtg cggattcacg gtcaataaag tatgtggttc gggtcttaaa agtgtggcgc 360

ttgccgccca ggccattcag gcaggtcagg cgcagagcat tgtggcgggg ggtatggaaa 420

atatgagttt agccccctac ttactcgatg caaaagcacg ctctggttat cgtcttggag 480

acggacaggt ttatgacgta atcctgcgcg atggcctgat gtgcgccacc catggttatc 540

atatggggat taccgccgaa aacgtggcta aagagtacgg aattacccgt gaaatgcagg 600

atgaactggc gctacattca cagcgtaaag cggcagccgc aattgagtcc ggtgctttta 660

cagccgaaat cgtcccggta aatgttgtca ctcgaaagaa aaccttcgtc ttcagtcaag 720

acgaattccc gaaagcgaat tcaacggctg aagcgttagg tgcattgcgc ccggccttcg 780

ataaagcagg aacagtcacc gctgggaacg cgtctggtat taacgacggt gctgccgctc 840

tggtgattat ggaagaatct gcggcgctgg cagcaggcct tacccccctg gctcgcatta 900

aaagttatgc cagcggtggc gtgccccccg cattgatggg tatggggcca gtacctgcca 960

cgcaaaaagc gttacaactg gcggggctgc aactggcgga tattgatctc attgaggcta 1020

atgaagcatt tgctgcacag ttccttgccg ttgggaaaaa cctgggcttt gattctgaga 1080

aagtgaatgt caacggcggg gccatcgcgc tcgggcatcc tatcggtgcc agtggtgctc 1140

gtattctggt cacactatta catgccatgc aggcacgcga taaaacgctg gggctggcaa 1200

cactgtgcat tggcggcggt cagggaattg cgatggtgat tgaacggttg aattaaagga 1260

ggaattaacc atgaaactct caactaaact ttgttggtgt ggtattaaag gaagacttag 1320

gccgcaaaag caacaacaat tacacaatac aaacttgcaa atgactgaac taaaaaaaca 1380

aaagaccgct gaacaaaaaa ccagacctca aaatgtcggt attaaaggta tccaaattta 1440

catcccaact caatgtgtca accaatctga gctagagaaa tttgatggcg tttctcaagg 1500

taaatacaca attggtctgg gccaaaccaa catgtctttt gtcaatgaca gagaagatat 1560

ctactcgatg tccctaactg ttttgtctaa gttgatcaag agttacaaca tcgacaccaa 1620

caaaattggt agattagaag tcggtactga aactctgatt gacaagtcca agtctgtcaa 1680

gtctgtcttg atgcaattgt ttggtgaaaa cactgacgtc gaaggtattg acacgcttaa 1740

tgcctgttac ggtggtacca acgcgttgtt caactctttg aactggattg aatctaacgc 1800

atgggatggt agagacgcca ttgtagtttg cggtgatatt gccatctacg ataagggtgc 1860

cgcaagacca accggtggtg ccggtactgt tgctatgtgg atcggtcctg atgctccaat 1920

tgtatttgac tctgtaagag cttcttacat ggaacacgcc tacgattttt acaagccaga 1980

tttcaccagc gaatatcctt acgtcgatgg tcatttttca ttaacttgtt acgtcaaggc 2040

tcttgatcaa gtttacaaga gttattccaa gaaggctatt tctaaagggt tggttagcga 2100

tcccgctggt tcggatgctt tgaacgtttt gaaatatttc gactacaacg ttttccatgt 2160

tccaacctgt aaattggtca caaaatcata cggtagatta ctatataacg atttcagagc 2220

caatcctcaa ttgttcccag aagttgacgc cgaattagct actcgcgatt atgacgaatc 2280

tttaaccgat aagaacattg aaaaaacttt tgttaatgtt gctaagccat tccacaaaga 2340

gagagttgcc caatctttga ttgttccaac aaacacaggt aacatgtaca ccgcatctgt 2400

ttatgccgcc tttgcatctc tattaaacta tgttggatct gacgacttac aaggcaagcg 2460

tgttggttta ttttcttacg gttccggttt agctgcatct ctatattctt gcaaaattgt 2520

tggtgacgtc caacatatta tcaaggaatt agatattact aacaaattag ccaagagaat 2580

caccgaaact ccaaaggatt acgaagctgc catcgaattg agagaaaatg cccatttgaa 2640

gaagaacttc aaacctcaag gttccattga gcatttgcaa agtggtgttt actacttgac 2700

caacatcgat gacaaattta gaagatctta cgatgttaaa aaataaagga ggaattaacc 2760

atgtcacttc ccttaaaaac gatagtacat ttggtaaagc cctttgcttg cactgctagg 2820

tttagtgcga gatacccaat ccacgtcatt gttgttgctg ttttattgag tgccgctgct 2880

tatctatccg tgacacaatc ttaccttaac gaatggaagc tggactctaa tcagtattct 2940

acatacttaa gcataaagcc ggatgagttg tttgaaaaat gcacacacta ctataggtct 3000

cctgtgtctg atacatggaa gttactcagc tctaaagaag ccgccgatat ttatacccct 3060

tttcattatt atttgtctac cataagtttt caaagtaagg acaattcaac gactttgcct 3120

tcccttgatg acgttattta cagtgttgac cataccaggt acttattaag tgaagagcca 3180

aagataccaa ctgaactagt gtctgaaaac ggaacgaaat ggagattgag aaacaacagc 3240

aattttattt tggacctgca taatatttac cgaaatatgg tgaagcaatt ttctaacaaa 3300

acgagcgaat ttgatcagtt cgatttgttt atcatcctag ctgcttacct tactcttttt 3360

tatactctct gttgcctgtt taatgacatg aggaaaatcg gatcaaagtt ttggttaagc 3420

ttttctgctc tttcaaactc tgcatgcgca ttatatttat cgctgtacac aactcacagt 3480

ttattgaaga aaccggcttc cttattaagt ttggtcattg gactaccatt tatcgtagta 3540

attattggct ttaagcataa agttcgactt gcggcattct cgctacaaaa attccacaga 3600

attagtattg acaagaaaat aacggtaagc aacattattt atgaggctat gtttcaagaa 3660

ggtgcctact taatccgcga ctacttattt tatattagct ccttcattgg atgtgctatt 3720

tatgctagac atcttcccgg attggtcaat ttctgtattt tgtctacatt tatgctagtt 3780

ttcgacttgc ttttgtctgc tactttttat tctgccattt tatcaatgaa gctggaaatt 3840

aacatcattc acagatcaac cgtcatcaga cagactttgg aagaggacgg agttgtccca 3900

actacagcag atattatata taaggatgaa actgcctcag aaccacattt tttgagatct 3960

aacgtggcta tcattctggg aaaagcatca gttattggtc ttttgcttct gatcaacctt 4020

tatgttttca cagataagtt aaatgctaca atactaaaca cggtatattt tgactctaca 4080

atttactcgt taccaaattt tatcaattat aaagatattg gcaatctcag caatcaagtg 4140

atcatttccg tgttgccaaa gcaatattat actccgctga aaaaatacca tcagatcgaa 4200

gattctgttc tacttatcat tgattccgtt agcaatgcta ttcgggacca atttatcagc 4260

aagttacttt tttttgcatt tgcagttagt atttccatca atgtctactt actgaatgct 4320

gcaaaaattc acacaggata catgaacttc caaccacaat caaataagat cgatgatctt 4380

gttgttcagc aaaaatcggc aacgattgag ttttcagaaa ctcgaagtat gcctgcttct 4440

tctggcctag aaactccagt gaccgcgaaa gatataatta tctctgaaga aatccagaat 4500

aacgaatgcg tctatgcttt gagttcccag gacgagccta tccgtccttt atcgaattta 4560

gtggaactta tggagaaaga acaattaaag aacatgaata atactgaggt ttcgaatctt 4620

gtcgtcaacg gtaaactgcc attatattcc ttagagaaaa aattagagga cacaactcgt 4680

gcggttttag ttaggagaaa ggcactttca actttggctg aatcgccaat tttagtttcc 4740

gaaaaattgc ccttcagaaa ttatgattat gatcgcgttt ttggagcttg ctgtgaaaat 4800

gtcatcggct atatgccaat accagttggt gtaattggtc cattaattat tgatggaaca 4860

tcttatcaca taccaatggc aaccacggaa ggttgtttag tggcttcagc tatgcgtggt 4920

tgcaaagcca tcaatgctgg tggtggtgca acaactgttt taaccaaaga tggtatgact 4980

agaggcccag tcgttcgttt ccctacttta ataagatctg gtgcctgcaa gatatggtta 5040

gactcggaag agggacaaaa ttcaattaaa aaagctttta attctacatc aaggtttgca 5100

cgtttgcaac atattcaaac ctgtctagca ggcgatttgc tttttatgag atttcggaca 5160

actaccggtg acgcaatggg tatgaacatg atatcgaaag gtgtcgaata ctctttgaaa 5220

caaatggtag aagaatatgg ttgggaagat atggaagttg tctccgtatc tggtaactat 5280

tgtactgata agaaacctgc cgcaatcaat tggattgaag gtcgtggtaa aagtgtcgta 5340

gctgaagcta ctattcctgg tgatgtcgta aaaagtgttt taaagagcga tgtttccgct 5400

ttagttgaat taaatatatc caagaacttg gttggatccg caatggctgg atctgttggt 5460

ggtttcaacg cgcacgcagc taatttggtc actgcacttt tcttggcatt aggccaagat 5520

cctgcgcaga acgtcgaaag ttccaactgt ataactttga tgaaggaagt tgatggtgat 5580

ttaaggatct ctgtttccat gccatctatt gaagttggta cgattggcgg gggtactgtt 5640

ctggagcctc agggcgccat gcttgatctt ctcggcgttc gtggtcctca ccccactgaa 5700

cctggagcaa atgctaggca attagctaga ataatcgcgt gtgctgtctt ggctggtgaa 5760

ctgtctctgt gctccgcact tgctgccggt cacctggtac aaagccatat gactcacaac 5820

cgtaaaacaa acaaagccaa tgaactgcca caaccaagta acaaagggcc cccctgtaaa 5880

acctcagcat tattataatg cttaagtcga acagaaagta atcgtattgt acacggccgc 5940

ataatcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca attccccatc 6000

ttagtatatt agttaagtat aagaaggaga tatacatatg gcagatctca attggatatc 6060

ggccggccac gcgatcgctg acgtcggtac cctcgagtct ggtaaagaaa ccgctgctgc 6120

gaaatttgaa cgccagcaca tggactcgtc tactagcgca gcttaattaa cctaggctgc 6180

tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg 6240

ttttttgctg aaacctcagg catttgagaa gcacacggtc acactgcttc cggtagtcaa 6300

taaaccggta aaccagcaat agacataagc ggctatttaa cgaccctgcc ctgaaccgac 6360

gaccgggtcg aatttgcttt cgaatttctg ccattcatcc gcttattatc acttattcag 6420

gcgtagcacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct 6480

gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac 6540

agacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat 6600

atttgcccat agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa 6660

aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt 6720

tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa 6780

actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat 6840

ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg 6900

ccatacggaa ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat 6960

aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg 7020

tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc 7080

attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag 7140

ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt 7200

gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg 7260

gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca 7320

caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta 7380

tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca 7440

gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt 7500

gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag 7560

gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc 7620

ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga 7680

acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc 7740

gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc 7800

tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tcccctggcg 7860

gctccctcgt gcgctctcct gttcctgcct ttcggtttac cggtgtcatt ccgctgttat 7920

ggccgcgttt gtctcattcc acgcctgaca ctcagttccg ggtaggcagt tcgctccaag 7980

ctggactgta tgcacgaacc ccccgttcag tccgaccgct gcgccttatc cggtaactat 8040

cgtcttgagt ccaacccgga aagacatgca aaagcaccac tggcagcagc cactggtaat 8100

tgatttagag gagttagtct tgaagtcatg cgccggttaa ggctaaactg aaaggacaag 8160

ttttggtgac tgcgctcctc caagccagtt acctcggttc aaagagttgg tagctcagag 8220

aaccttcgaa aaaccgccct gcaaggcggt tttttcgttt tcagagcaag agattacgcg 8280

cagaccaaaa cgatctcaag aagatcatct tattaatcag ataaaatatt tctagatttc 8340

agtgcaattt atctcttcaa atgtagcacc tgaagtcagc cccatacgat ataagttgta 8400

attctcatgt tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt gaaggctctc 8460

aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt aattgcgttg 8520

cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc 8580

caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc ttttcaccag 8640

tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt gcagcaagcg 8700

gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta acggcgggat 8760

ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg caccaacgcg 8820

cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt tggcaaccag 8880

catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa aaccggacat 8940

ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag tgagatattt 9000

atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg ctaacagcgc 9060

gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac cgtcttcatg 9120

ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata acgccggaac 9180

attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat agttaatgat 9240

cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg cttcgacgcc 9300

gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc gagatttaat 9360

cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa cgccaatcag 9420

caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat tcagctccgc 9480

catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct ggttcaccac 9540

gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata acgttactgg 9600

tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca taccgcgaaa 9660

ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc gactcctgca 9720

ttaggaaatt aatacgactc actata 9746

<210> 5

<211> 11133

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac 60

tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca 120

ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta 180

aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata 240

ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag 300

cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag 360

caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg 420

tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct 480

tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc 540

ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc 600

gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca 660

tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga 720

tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa 780

acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata 840

taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc 900

ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt 960

tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat 1020

tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta 1080

accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt 1140

aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca 1200

ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta 1260

tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattaacca 1320

tgaaaaattg tgtcatcgtc agtgcggtac gtactgctat cggtagtttt aacggttcac 1380

tcgcttccac cagcgccatc gacctggggg cgacagtaat taaagccgcc attgaacgtg 1440

caaaaatcga ttcacaacac gttgatgaag tgattatggg taacgtgtta caagccgggc 1500

tggggcaaaa tccggcgcgt caggcactgt taaaaagcgg gctggcagaa acggtgtgcg 1560

gattcacggt caataaagta tgtggttcgg gtcttaaaag tgtggcgctt gccgcccagg 1620

ccattcaggc aggtcaggcg cagagcattg tggcgggggg tatggaaaat atgagtttag 1680

ccccctactt actcgatgca aaagcacgct ctggttatcg tcttggagac ggacaggttt 1740

atgacgtaat cctgcgcgat ggcctgatgt gcgccaccca tggttatcat atggggatta 1800

ccgccgaaaa cgtggctaaa gagtacggaa ttacccgtga aatgcaggat gaactggcgc 1860

tacattcaca gcgtaaagcg gcagccgcaa ttgagtccgg tgcttttaca gccgaaatcg 1920

tcccggtaaa tgttgtcact cgaaagaaaa ccttcgtctt cagtcaagac gaattcccga 1980

aagcgaattc aacggctgaa gcgttaggtg cattgcgccc ggccttcgat aaagcaggaa 2040

cagtcaccgc tgggaacgcg tctggtatta acgacggtgc tgccgctctg gtgattatgg 2100

aagaatctgc ggcgctggca gcaggcctta cccccctggc tcgcattaaa agttatgcca 2160

gcggtggcgt gccccccgca ttgatgggta tggggccagt acctgccacg caaaaagcgt 2220

tacaactggc ggggctgcaa ctggcggata ttgatctcat tgaggctaat gaagcatttg 2280

ctgcacagtt ccttgccgtt gggaaaaacc tgggctttga ttctgagaaa gtgaatgtca 2340

acggcggggc catcgcgctc gggcatccta tcggtgccag tggtgctcgt attctggtca 2400

cactattaca tgccatgcag gcacgcgata aaacgctggg gctggcaaca ctgtgcattg 2460

gcggcggtca gggaattgcg atggtgattg aacggttgaa ttaaaggagg aattaaccat 2520

gaaactctca actaaacttt gttggtgtgg tattaaagga agacttaggc cgcaaaagca 2580

acaacaatta cacaatacaa acttgcaaat gactgaacta aaaaaacaaa agaccgctga 2640

acaaaaaacc agacctcaaa atgtcggtat taaaggtatc caaatttaca tcccaactca 2700

atgtgtcaac caatctgagc tagagaaatt tgatggcgtt tctcaaggta aatacacaat 2760

tggtctgggc caaaccaaca tgtcttttgt caatgacaga gaagatatct actcgatgtc 2820

cctaactgtt ttgtctaagt tgatcaagag ttacaacatc gacaccaaca aaattggtag 2880

attagaagtc ggtactgaaa ctctgattga caagtccaag tctgtcaagt ctgtcttgat 2940

gcaattgttt ggtgaaaaca ctgacgtcga aggtattgac acgcttaatg cctgttacgg 3000

tggtaccaac gcgttgttca actctttgaa ctggattgaa tctaacgcat gggatggtag 3060

agacgccatt gtagtttgcg gtgatattgc catctacgat aagggtgccg caagaccaac 3120

cggtggtgcc ggtactgttg ctatgtggat cggtcctgat gctccaattg tatttgactc 3180

tgtaagagct tcttacatgg aacacgccta cgatttttac aagccagatt tcaccagcga 3240

atatccttac gtcgatggtc atttttcatt aacttgttac gtcaaggctc ttgatcaagt 3300

ttacaagagt tattccaaga aggctatttc taaagggttg gttagcgatc ccgctggttc 3360

ggatgctttg aacgttttga aatatttcga ctacaacgtt ttccatgttc caacctgtaa 3420

attggtcaca aaatcatacg gtagattact atataacgat ttcagagcca atcctcaatt 3480

gttcccagaa gttgacgccg aattagctac tcgcgattat gacgaatctt taaccgataa 3540

gaacattgaa aaaacttttg ttaatgttgc taagccattc cacaaagaga gagttgccca 3600

atctttgatt gttccaacaa acacaggtaa catgtacacc gcatctgttt atgccgcctt 3660

tgcatctcta ttaaactatg ttggatctga cgacttacaa ggcaagcgtg ttggtttatt 3720

ttcttacggt tccggtttag ctgcatctct atattcttgc aaaattgttg gtgacgtcca 3780

acatattatc aaggaattag atattactaa caaattagcc aagagaatca ccgaaactcc 3840

aaaggattac gaagctgcca tcgaattgag agaaaatgcc catttgaaga agaacttcaa 3900

acctcaaggt tccattgagc atttgcaaag tggtgtttac tacttgacca acatcgatga 3960

caaatttaga agatcttacg atgttaaaaa ataaaggagg aattaaccat gtcacttccc 4020

ttaaaaacga tagtacattt ggtaaagccc tttgcttgca ctgctaggtt tagtgcgaga 4080

tacccaatcc acgtcattgt tgttgctgtt ttattgagtg ccgctgctta tctatccgtg 4140

acacaatctt accttaacga atggaagctg gactctaatc agtattctac atacttaagc 4200

ataaagccgg atgagttgtt tgaaaaatgc acacactact ataggtctcc tgtgtctgat 4260

acatggaagt tactcagctc taaagaagcc gccgatattt ataccccttt tcattattat 4320

ttgtctacca taagttttca aagtaaggac aattcaacga ctttgccttc ccttgatgac 4380

gttatttaca gtgttgacca taccaggtac ttattaagtg aagagccaaa gataccaact 4440

gaactagtgt ctgaaaacgg aacgaaatgg agattgagaa acaacagcaa ttttattttg 4500

gacctgcata atatttaccg aaatatggtg aagcaatttt ctaacaaaac gagcgaattt 4560

gatcagttcg atttgtttat catcctagct gcttacctta ctctttttta tactctctgt 4620

tgcctgttta atgacatgag gaaaatcgga tcaaagtttt ggttaagctt ttctgctctt 4680

tcaaactctg catgcgcatt atatttatcg ctgtacacaa ctcacagttt attgaagaaa 4740

ccggcttcct tattaagttt ggtcattgga ctaccattta tcgtagtaat tattggcttt 4800

aagcataaag ttcgacttgc ggcattctcg ctacaaaaat tccacagaat tagtattgac 4860

aagaaaataa cggtaagcaa cattatttat gaggctatgt ttcaagaagg tgcctactta 4920

atccgcgact acttatttta tattagctcc ttcattggat gtgctattta tgctagacat 4980

cttcccggat tggtcaattt ctgtattttg tctacattta tgctagtttt cgacttgctt 5040

ttgtctgcta ctttttattc tgccatttta tcaatgaagc tggaaattaa catcattcac 5100

agatcaaccg tcatcagaca gactttggaa gaggacggag ttgtcccaac tacagcagat 5160

attatatata aggatgaaac tgcctcagaa ccacattttt tgagatctaa cgtggctatc 5220

attctgggaa aagcatcagt tattggtctt ttgcttctga tcaaccttta tgttttcaca 5280

gataagttaa atgctacaat actaaacacg gtatattttg actctacaat ttactcgtta 5340

ccaaatttta tcaattataa agatattggc aatctcagca atcaagtgat catttccgtg 5400

ttgccaaagc aatattatac tccgctgaaa aaataccatc agatcgaaga ttctgttcta 5460

cttatcattg attccgttag caatgctatt cgggaccaat ttatcagcaa gttacttttt 5520

tttgcatttg cagttagtat ttccatcaat gtctacttac tgaatgctgc aaaaattcac 5580

acaggataca tgaacttcca accacaatca aataagatcg atgatcttgt tgttcagcaa 5640

aaatcggcaa cgattgagtt ttcagaaact cgaagtatgc ctgcttcttc tggcctagaa 5700

actccagtga ccgcgaaaga tataattatc tctgaagaaa tccagaataa cgaatgcgtc 5760

tatgctttga gttcccagga cgagcctatc cgtcctttat cgaatttagt ggaacttatg 5820

gagaaagaac aattaaagaa catgaataat actgaggttt cgaatcttgt cgtcaacggt 5880

aaactgccat tatattcctt agagaaaaaa ttagaggaca caactcgtgc ggttttagtt 5940

aggagaaagg cactttcaac tttggctgaa tcgccaattt tagtttccga aaaattgccc 6000

ttcagaaatt atgattatga tcgcgttttt ggagcttgct gtgaaaatgt catcggctat 6060

atgccaatac cagttggtgt aattggtcca ttaattattg atggaacatc ttatcacata 6120

ccaatggcaa ccacggaagg ttgtttagtg gcttcagcta tgcgtggttg caaagccatc 6180

aatgctggtg gtggtgcaac aactgtttta accaaagatg gtatgactag aggcccagtc 6240

gttcgtttcc ctactttaat aagatctggt gcctgcaaga tatggttaga ctcggaagag 6300

ggacaaaatt caattaaaaa agcttttaat tctacatcaa ggtttgcacg tttgcaacat 6360

attcaaacct gtctagcagg cgatttgctt tttatgagat ttcggacaac taccggtgac 6420

gcaatgggta tgaacatgat atcgaaaggt gtcgaatact ctttgaaaca aatggtagaa 6480

gaatatggtt gggaagatat ggaagttgtc tccgtatctg gtaactattg tactgataag 6540

aaacctgccg caatcaattg gattgaaggt cgtggtaaaa gtgtcgtagc tgaagctact 6600

attcctggtg atgtcgtaaa aagtgtttta aagagcgatg tttccgcttt agttgaatta 6660

aatatatcca agaacttggt tggatccgca atggctggat ctgttggtgg tttcaacgcg 6720

cacgcagcta atttggtcac tgcacttttc ttggcattag gccaagatcc tgcgcagaac 6780

gtcgaaagtt ccaactgtat aactttgatg aaggaagttg atggtgattt aaggatctct 6840

gtttccatgc catctattga agttggtacg attggcgggg gtactgttct ggagcctcag 6900

ggcgccatgc ttgatcttct cggcgttcgt ggtcctcacc ccactgaacc tggagcaaat 6960

gctaggcaat tagctagaat aatcgcgtgt gctgtcttgg ctggtgaact gtctctgtgc 7020

tccgcacttg ctgccggtca cctggtacaa agccatatga ctcacaaccg taaaacaaac 7080

aaagccaatg aactgccaca accaagtaac aaagggcccc cctgtaaaac ctcagcatta 7140

ttataatgtt ttggcggatg agagaagatt ttcagcctga tacagattaa atcagaacgc 7200

agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt cccacctgac 7260

cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg gtctccccat 7320

gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc 7380

ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg 7440

agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata 7500

aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct 7560

acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 7620

aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 7680

gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 7740

cgctggtgaa agtaaaagat gctgaagatc agttggggca aactattaac tggcgaacta 7800

cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga 7860

ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt 7920

gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc 7980

gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct 8040

gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata 8100

ctttagattg atttacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 8160

gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 8220

ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 8280

agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg 8340

ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 8400

gttctttaat agtggactct tgttccaaac ttgaacaaca ctcaacccta tctcgggcta 8460

ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 8520

ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt aaaaggatct 8580

aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc 8640

actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc 8700

gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg 8760

atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa 8820

atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc 8880

ctacatacct cgctctgcta atcctgttac cagtcaggca tttgagaagc acacggtcac 8940

actgcttccg gtagtcaata aaccggtaaa ccagcaatag acataagcgg ctatttaacg 9000

accctgccct gaaccgacga ccgggtcgaa tttgctttcg aatttctgcc attcatccgc 9060

ttattatcac ttattcaggc gtagcaccag gcgtttaagg gcaccaataa ctgccttaaa 9120

aaaattacgc cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc 9180

cgacatggaa gccatcacag acggcatgat gaacctgaat cgccagcggc atcagcacct 9240

tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat 9300

tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca 9360

tattctcaat aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt 9420

gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa 9480

acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca 9540

gctcaccgtc tttcattgcc atacggaatt ccggatgagc attcatcagg cgggcaagaa 9600

tgtgaataaa ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg 9660

taatatccag ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa 9720

aatgttcttt acgatgccat tgggatatat caacggtggt atatccagtg atttttttct 9780

ccattttagc ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg 9840

atcttatttc attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattttc 9900

gccaaaagtt ggcccagggc ttcccggtat caacagggac accaggattt atttattctg 9960

cgaagtgatc ttccgtcaca ggtatttatt cggcgcaaag tgcgtcgggt gatgctgcca 10020

acttactgat ttagtgtatg atggtgtttt tgaggtgctc cagtggcttc tgtttctatc 10080

agctgtccct cctgttcagc tactgacggg gtggtgcgta acggcaaaag caccgccgga 10140

catcagcgct agcggagtgt atactggctt actatgttgg cactgatgag ggtgtcagtg 10200

aagtgcttca tgtggcagga gaaaaaaggc tgcaccggtg cgtcagcaga atatgtgata 10260

caggatatat tccgcttcct cgctcactga ctcgctacgc tcggtcgttc gactgcggcg 10320

agcggaaatg gcttacgaac ggggcggaga tttcctggaa gatgccagga agatacttaa 10380

cagggaagtg agagggccgc ggcaaagccg tttttccata ggctccgccc ccctgacaag 10440

catcacgaaa tctgacgctc aaatcagtgg tggcgaaacc cgacaggact ataaagatac 10500

caggcgtttc cccctggcgg ctccctcgtg cgctctcctg ttcctgcctt tcggtttacc 10560

ggtgtcattc cgctgttatg gccgcgtttg tctcattcca cgcctgacac tcagttccgg 10620

gtaggcagtt cgctccaagc tggactgtat gcacgaaccc cccgttcagt ccgaccgctg 10680

cgccttatcc ggtaactatc gtcttgagtc caacccggaa agacatgcaa aagcaccact 10740

ggcagcagcc actggtaatt gatttagagg agttagtctt gaagtcatgc gccggttaag 10800

gctaaactga aaggacaagt tttggtgact gcgctcctcc aagccagtta cctcggttca 10860

aagagttggt agctcagaga accttcgaaa aaccgccctg caaggcggtt ttttcgtttt 10920

cagagcaaga gattacgcgc agaccaaaac gatctcaaga agatcatctt attaatcaga 10980

taaaatattt gctcatgagc ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg 11040

cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg atgcgtccgg 11100

cgtagaggat ctgctcatgt ttgacagctt atc 11133

<210> 6

<211> 6915

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 60

cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 120

ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 180

gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 240

acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 300

ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 360

ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 420

acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 480

tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 540

tccgctcatg aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat 600

tcatatcagg attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa 660

actcaccgag gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc 720

gtccaacatc aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga 780

aatcaccatg agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc 840

agacttgttc aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac 900

cgttattcat tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac 960

aattacaaac aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat 1020

tttcacctga atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag 1080

tggtgagtaa ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca 1140

taaattccgt cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac 1200

ctttgccatg tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg 1260

tcgcacctga ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca 1320

tgttggaatt taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac 1380

cccttgtatt actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa 1440

cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga 1500

gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg 1560

gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc 1620

agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag 1680

aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc 1740

agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg 1800

cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac 1860

accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga 1920

aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt 1980

ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag 2040

cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg 2100

gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta 2160

tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc 2220

agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg 2280

tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatatggtgc actctcagta 2340

caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg 2400

ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 2460

gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 2520

gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc 2580

gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag 2640

aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt 2700

ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatgaa 2760

acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt tactggaacg 2820

ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa tcactcaggg 2880

tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc agcagcatcc 2940

tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt ccagacttta 3000

cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg ttttgcagca 3060

gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag taaggcaacc 3120

ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc gtggggccgc 3180

catgccggcg ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa 3240

ggcttgagcg agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc 3300

gctccagcga aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac 3360

gagttgcatg ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca 3420

ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta 3480

atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa 3540

cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat 3600

tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca 3660

ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa 3720

aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt 3780

atcccactac cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg 3840

cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca 3900

gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta 3960

tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg 4020

agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat 4080

gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct 4140

ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg 4200

catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat 4260

tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc 4320

tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca 4380

gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg 4440

ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt 4500

tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg 4560

catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct 4620

cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga 4680

tctcgacgct ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg 4740

ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc 4800

ccggccacgg ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg 4860

cgagcccgat cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg 4920

gcgccggtga tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga 4980

aattaatacg actcactata ggggaattgt gagcggataa caattcccct ctagaaataa 5040

ttttgtttaa ctttaggagg aattaaccat gtcgactctt cctatttcta gtgtttcatt 5100

ctcttcatct acgtcacctt tggtagttga tgataaagtt agcacgaaac aagatgttat 5160

ccgccatacc atgaatttca atgctagcat atggggagat caatttctta catatgatga 5220

gccagaagat ttggttatga agaaacaact tgttgaagag ctcaaagagg aagtcaagaa 5280

agagctaata acgatcaaag gttctaatga accgatgcag catgtcaagt tgttagaact 5340

tattgatgca gtccaacggc ttggcatagc ctatcatttt gaagaggaga ttgaagaagc 5400

cttgcaacat atccatgtta catatggcga gcagtgggtc gataaagaaa acctacaaag 5460

catttctctt tggttccgac tcctacgaca acaaggcttc aacgtctcat caggagtatt 5520

caaggacttt atggacgaga agggaaaatt taaggaatcc ttatgtaatg atgctcaggg 5580

aattcttgct ctgtatgaag cggcatttat gagggtggaa gatgaaacaa tactagataa 5640

tgcgctcgag ttcactaaag ttcaccttga catcatagcc aaggatcctt cttgtgactc 5700

ttccctaaga acccaaatac accaagcgtt aaagcagcca cttaggagaa ggctggcaag 5760

gatagaggcg ttgcattaca tgcctatcta ccaacaagaa acatcccaca atgaggtctt 5820

actgaagctt gcaaaactag attttagcgt gcttcaatca atgcacaaaa aggagcttag 5880

ccacatttgc aaatggtgga aggatttgga ccttcaaaat aagcttcctt atgttcgaga 5940

cagggtggtg gaaggctatt tttggatact atccatctat tacgagcctc aacatgctcg 6000

aacaagaatg ttcctaatga aaacatgcat gtggttagtc gttttagatg atacatttga 6060

taattatggt acttatgaag aactcgaaat ctttacacaa gctgttgaaa gatggtcaat 6120

aagctgcctg gatatgcttc cagaatacat gaaactaata tatcaagagc ttgtgaatct 6180

tcacgtggaa atggaggaat cacttgaaaa ggagggaaaa acatatcaaa ttcactatgt 6240

caaggagatg gcaaaagagt tggttcgcaa ctacttggta gaagcgagat ggctaaaaga 6300

ggggtacatg ccaactcttg aggagtacat gtctgtgtca atggtgactg gtacctatgg 6360

cttaatgata gcgagatctt atgtcggtag gggtgatatt gtcaccgagg atacctttaa 6420

atgggtgtcc tcgtatcctc ctattataaa agcttcatgt gtgattgtaa gacttatgga 6480

tgacattgtc agccacaagg aggaacaaga gagaggccat gttgcttcaa gcatcgaatg 6540

ctattctaag gaaagtggtg catcagagga ggaagcgtgt gaatatatct caggaaaagt 6600

tgaagatgca tggaaagtta taaaccgaga gtcgctcagg cctacagcgg tcccgtttcc 6660

tctacttatg cctgcaatca accttgcacg tatgtgtgaa gtcctatata gcgtcaacga 6720

tggcttcact catgctgagg gagacatgaa aagttacatg aaatcgttct tcgttcaccc 6780

tatggttgtc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat 6840

aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag 6900

gaactatatc cggat 6915

Claims (6)

1. An engineering strain is characterized in that the engineering strain is escherichia coli with the preservation number of CGMCC NO. 14818.

2. Use of the engineered strain of claim 1 to produce farnesene by fermentation with a cellulose hydrolysate.

3. A method for preparing farnesene by using cellulose is characterized by comprising the following steps:

(1) hydrolyzing a solid raw material containing cellulose to obtain a hydrolysate containing monosaccharide;

(2) inoculating the engineering strain of claim 1 into a liquid fermentation raw material containing the hydrolysate containing monosaccharide, and performing fermentation culture to obtain a cultured material.

4. The method of claim 3, wherein,

the step of subjecting the cellulose-containing raw material to hydrolysis treatment comprises:

s1, carrying out acid pretreatment and alkali pretreatment on the solid raw material containing cellulose in sequence, or carrying out alkali pretreatment and acid pretreatment in sequence;

s2, carrying out enzymatic hydrolysis on the pretreated solid material;

the acid pretreatment is formic acid treatment and/or dilute sulfuric acid treatment; the formic acid treatment conditions include: the temperature is 40-100 ℃, the time is 1-10 hours, the volume of a formic acid solution is 5-50 liters per kilogram of pretreated solid material, and the formic acid solution contains 70-95 wt% of formic acid and 0-2 wt% of HCl; the conditions of the dilute sulfuric acid treatment include: the temperature is 80-150 ℃, the time is 5 minutes-2 hours, and the volume of the dilute sulfuric acid solution is 5-50 liters per kilogram of pretreated solid material, wherein the dilute sulfuric acid solution contains 0.5-10 wt% of dilute sulfuric acid;

the alkali pretreatment is ammonia water treatment and/or sodium hydroxide treatment; the ammonia water treatment conditions comprise: the temperature is 40-80 ℃, the time is 2-24 hours, the volume of ammonia water is 2-20 liters per kilogram of pretreated solid material, and the ammonia water contains 10-25 wt% of NH₃(ii) a The sodium hydroxide treatment conditions include: the temperature is 60-120 ℃, the time is 1-12 hours, and the volume of a sodium hydroxide aqueous solution is 5-50 liters per kilogram of pretreated solid material, wherein the sodium hydroxide aqueous solution contains 0.5-10 wt% of sodium hydroxide;

the conditions of the enzymatic hydrolysis include: the temperature is 30-70 ℃, the time is 3-72 hours, the pH is 4.0-6.0, the volume of a buffer solution is 5-100 liters per kilogram of pretreated solid materials, the buffer solution used for enzymatic hydrolysis contains 0.01-1 mol/L citric acid buffer solution and/or 0.01-1 mol/L acetic acid buffer solution, the enzyme used for enzymatic hydrolysis is cellulase and/or beta-glucosidase, the enzyme adding amount of the cellulase is 10-500U per gram of pretreated solid materials, and the enzyme adding amount of the beta-glucosidase is 10-250U per gram of pretreated solid materials.

5. The method of claim 3 or 4, wherein the conditions of the fermentation culture comprise: the temperature is 16-35 ℃, the time is 3-120 h, the dosage of the extracting agent is 50-1000 mL relative to each liter of the liquid fermentation raw material, and the inoculation amount of the engineering bacteria or the engineering strains is (10-300) multiplied by 10⁷A plurality of; the extractant is selected from n-hexane, n-decane, dodecane or kerosene; and in each liter of the liquid fermentation raw material, the content of the hydrolysate containing monosaccharide is 0.01-0.9 kg calculated by the amount of monosaccharide.

6. The method of claim 3, wherein the cellulose-containing feedstock comprises at least one of straw, bark, wood chips, corn cobs, rice bran, and wheat bran.