CN109722433B - Application of nucleic acid for encoding NhaA protein and method for preparing farnesene by using cellulose - Google Patents

Abstract

The present disclosure provides the use of nucleic acid encoding a NhaA protein in the preparation of an engineered bacterium that produces farnesene. The disclosure also provides a method for preparing the Albizzia julibrissin production engineering bacteria. The disclosure also provides the engineering bacteria prepared by the method. 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 bacteria into the liquid fermentation raw material containing the hydrolysate of monosaccharide, and performing fermentation culture to obtain a cultured material. Therefore, the present disclosure significantly improves the yield of farnesene produced by fermentation using cellulose hydrolysate according to the above technical solution.

Description

Application of nucleic acid for encoding NhaA protein and method for preparing farnesene by using cellulose

Technical Field

The disclosure relates to the field of bioengineering, in particular to application of nucleic acid for encoding NhaA protein in preparation of an engineered bacterium for producing farnesene, a method for preparing the engineered bacterium for producing the farnesene, the engineered bacterium prepared by the method and a method for preparing the 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 a use of a nucleic acid encoding a NhaA protein in the preparation of an engineered bacterium producing farnesene, wherein the NhaA protein is a protein of (a) or (b): (a) a protein consisting of an amino acid sequence shown as SEQ ID NO.1, and (b) a protein which is derived from (a) and has the activity of a NhaA protein, wherein one or more amino acids are substituted, deleted or added in the amino acid sequence in (a).

In another aspect, the present disclosure also provides a method for preparing an engineered bacterium for producing ajoene, wherein the method includes: the transformed host bacteria of the engineering bacteria can express isopentenyl pyrophosphate isomerase (IDI), farnesene pyrophosphate synthetase (ispA) and farnesene synthetase (FG) by transforming the host bacteria of the engineering bacteria into recombinant plasmids or integrating and knocking in chromosomes, and can express a plurality of enzymes forming a Mevalonate (MVA) pathway or a plurality of enzymes forming a deoxyxylulose-5-phosphate (MEP) pathway and can over-express NhaA protein; the NhaA protein is a protein of the following (a) or (b): (a) a protein consisting of an amino acid sequence shown as SEQ ID NO.1, and (b) a protein which is derived from (a) and has the activity of a NhaA protein, wherein one or more amino acids are substituted, deleted or added in the amino acid sequence in (a).

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

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 bacteria into the liquid fermentation raw material containing the hydrolysate of monosaccharide, and performing fermentation culture to obtain a cultured material.

The reason that the yield is low when farnesene is produced by fermenting the cellulose hydrolysate is probably that factors inhibiting fermentation exist in the cellulose hydrolysate, and the stress resistance of engineering bacteria can be remarkably improved by the over-expression of the NhaA protein. 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.

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 a use of a nucleic acid encoding a NhaA protein in preparing an engineered bacterium producing farnesene, wherein the NhaA protein is a protein of (a) or (b): (a) a protein consisting of an amino acid sequence shown as SEQ ID NO.1, and (b) a protein which is derived from (a) and has the activity of a NhaA protein, wherein one or more amino acids are substituted, deleted or added in the amino acid sequence in (a).

Wherein the NhaA protein activity refers to Na⁺/H⁺Activity of an ion antiporter; the method for detecting the activity of the NhaA protein may be a known method. Wherein the number of amino acids substituted, deleted or added in the amino acid sequence in (a) may be any number of 1 to 100.

Wherein, the nucleic acid for coding the NhaA protein can be a plurality of nucleic acids according to the degeneracy of codons, and the nucleotide sequence of the nucleic acid for coding the NhaA protein can be optimized according to the codon bias under the condition that the amino acid sequence of the coded NhaA protein is not changed.

Optionally, wherein the nucleic acid encoding the NhaA protein is a nucleic acid consisting of the nucleotide sequence shown in SEQ ID No. 2.

Optionally, the farnesene-producing engineering bacteria is engineering bacteria capable of producing farnesene by fermentation of a cellulose hydrolysate.

Optionally, wherein the engineering bacterium is escherichia coli.

Optionally, wherein the engineered bacteria capable of producing farnesene by fermentation with a cellulose hydrolysate can have expression of isopentenyl pyrophosphate isomerase (IDI), farnesene pyrophosphate synthetase (ispA) and farnesene synthetase (FG), and can have expression of a plurality of enzymes constituting a Mevalonate (MVA) pathway or a plurality of enzymes constituting a deoxyxylulose-5-phosphate (MEP) pathway.

Optionally, the plurality of enzymes comprising a Mevalonate (MVA) pathway comprises 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 and/or an MvaE enzyme; the HMG-CoA synthase is HMGS enzyme and/or MvaS enzyme, and the HMG-CoA reductase is HMGR enzyme and/or MvaE enzyme.

Alternatively, the plurality of enzymes constituting the deoxyxylulose-5-phosphate (MEP) pathway include 1-deoxy-D-xylulose-5-phosphate synthase (DXS), 1-deoxy-D-xylulose-5-phosphate reductoisomerase (IspC), cytidine-2C-methyl-D-erythritol 4-diphosphate synthase (IspD), cytidine-2C-methyl-D-erythritol 4-diphosphate synthase (IspE), 2C-methyl-D-erythrose-2, 4-cyclodiphosphate synthase (IspF), 1-hydroxy-2-methyl-2- (E) -butenyl 4-diphosphate synthase (IspG), and isopentenyl/dimethylallyl diphosphate synthase (ispH).

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

In another aspect, the present disclosure provides a method for preparing an engineered bacterium producing farnesene, wherein the method comprises: the transformed host bacteria of the engineering bacteria can express isopentenyl pyrophosphate isomerase (IDI), farnesene pyrophosphate synthetase (ispA) and farnesene synthetase (FG) by transforming the host bacteria of the engineering bacteria into recombinant plasmids or integrating and knocking in chromosomes, and can express a plurality of enzymes forming a Mevalonate (MVA) pathway or a plurality of enzymes forming a deoxyxylulose-5-phosphate (MEP) pathway and can over-express NhaA protein; the NhaA protein is a protein of the following (a) or (b): (a) a protein consisting of an amino acid sequence shown as SEQ ID NO.1, and (b) a protein which is derived from (a) and has the activity of a NhaA protein, wherein one or more amino acids are substituted, deleted or added in the amino acid sequence in (a).

In a preferred embodiment, the engineered bacteria host bacteria can be transformed by transferring recombinant plasmids or integrating and knocking in chromosomes, so that the transformed engineered bacteria host bacteria can express citrate lyase (ACL) and/or acetate kinase (ack). In this preferred embodiment, citrate lyase and acetate kinase can make citric acid and acetic acid as buffer components available to the fermenting engineering bacteria, thereby further improving the yield of farnesene.

Optionally, wherein the engineering bacteria is escherichia coli; the plurality of enzymes comprising a Mevalonate (MVA) pathway include 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 and/or an MvaE enzyme; the HMG-CoA synthase is HMGS enzyme and/or MvaS enzyme, and the HMG-CoA reductase is HMGR enzyme and/or MvaE enzyme.

Alternatively, the plurality of enzymes constituting the deoxyxylulose-5-phosphate (MEP) pathway include 1-deoxy-D-xylulose-5-phosphate synthase (DXS), 1-deoxy-D-xylulose-5-phosphate reductoisomerase (IspC), cytidine-2C-methyl-D-erythritol 4-diphosphate synthase (IspD), cytidine-2C-methyl-D-erythritol 4-diphosphate synthase (IspE), 2C-methyl-D-erythrose-2, 4-cyclodiphosphate synthase (IspF), 1-hydroxy-2-methyl-2- (E) -butenyl 4-diphosphate synthase (IspG), and isopentenyl/dimethylallyl diphosphate synthase (ispH).

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

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 bacteria into the liquid fermentation raw material containing the hydrolysate of 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: at a temperature ofThe 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 is (10-300) x 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 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, firstly 1 set of MEP metabolic pathway plasmids are constructed, specifically including pDCDE plasmid, pFGHII plasmid and pFG plasmid, then 1 set of MVA metabolic pathway plasmids are constructed, specifically including pMevT plasmid, pMBIS plasmid and pFG plasmid, and then 1 plasmid for replacing pFG plasmid and inserting NhaA gene is constructed, namely pFGN plasmid. 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 method into plasmid pDCDE, pFGHII and pFGN. The strain can synthesize farnesene through an MEP metabolic pathway and has over-expression of NhaA protein. On the antibiotic resistance selection plate, 3 clones of the strain were selected, F1-1, F1-2 and F1-3, respectively.

Construction of experimental group engineered strain F2: selecting Escherichia coli BL21 as host cell, passing through CaCl₂Transformation methods were transformed with plasmids pMevT, pMBIS and pFGN. The strain can synthesize farnesene through an MVA metabolic pathway and has over-expression of NhaA protein. On the antibiotic resistance selection plate, 3 clones of the strain were selected, F2-1, F2-2 and F2-3, respectively.

Construction of control engineered strain DF 1: selecting Escherichia coli BL21 as host cell, passing through CaCl₂Transformation method into plasmids pDCDE, pFGHII and pFGG. The strain can synthesize farnesene through an MEP metabolic pathway, but does not over-express the NhaA protein. Selecting the bacteria on antibiotic resistance screening plate3 clones of the strain are 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 method into plasmid pMevT, pMBIS and pFG. The strain can synthesize farnesene through an MVA metabolic pathway, but does not over-express NhaA protein. On the antibiotic resistance selection plate, 3 clones of the strain were selected, DF1-1, DF1-2 and DF1-3, 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/L) were added to the above liquid fermentation raw materials, and then 12 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, 3 clones F1-1, F1-2 and F1-3 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, while 3 clones F2-1, F2-2 and F2-3 of the experimental engineered strain F2 were all significantly higher than 3 clones DF2-1, DF2-2 and DF2-3 of the control engineered strain DF2 with respect to the farnesene content in the fermentation product. Therefore, the high expression of the NhaA protein can obviously improve the yield of farnesene produced by fermentation of a cellulose hydrolysate.

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

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Research Institute of petrochemicals, Sinopec

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tggcgctgca ctatgtctac aacaccccgt ttgaccaatt gatttgggat gtggggcatc 1560

aggcttatcc gcataaaatt ttgaccggac gccgcgacaa aatcggcacc atccgtcaga 1620

aaggcggtct gcacccgttc ccgtggcgcg gcgaaagcga atatgacgta ttaagcgtcg 1680

ggcattcatc aacctccatc agtgccggaa ttggtattgc ggttgctgcc gaaaaagaag 1740

gcaaaaatcg ccgcaccgtc tgtgtcattg gcgatggcgc gattaccgca ggcatggcgt 1800

ttgaagcgat gaatcacgcg ggcgatatcc gtcctgatat gctggtgatt ctcaacgaca 1860

atgaaatgtc gatttccgaa aatgtcggcg cgctcaacaa ccatctggca cagctgcttt 1920

ccggtaagct ttactcttca ctgcgcgaag gcgggaaaaa agttttctct ggcgtgccgc 1980

caattaaaga gctgctcaaa cgcaccgaag aacatattaa aggcatggta gtgcctggca 2040

cgttgtttga agagctgggc tttaactaca tcggcccggt ggacggtcac gatgtgctgg 2100

ggcttatcac cacgctaaag aacatgcgcg acctgaaagg cccgcagttc ctgcatatca 2160

tgaccaaaaa aggtcgtggt tatgaaccgg cagaaaaaga cccgatcact ttccacgccg 2220

tgcctaaatt tgatccctcc agcggttgtt tgccgaaaag tagcggcggt ttgccgagct 2280

attcaaaaat ctttggcgac tggttgtgcg aaacggcagc gaaagacaac aagctgatgg 2340

cgattactcc ggcgatgcgt gaaggttccg gcatggtcga gttttcacgt aaattcccgg 2400

atcgctactt cgacgtggca attgccgagc aacacgcggt gacctttgct gcgggtctgg 2460

cgattggtgg gtacaaaccc attgtcgcga tttactccac tttcctgcaa cgcgcctatg 2520

atcaggtgct gcatgacgtg gcgattcaaa agcttccggt cctgttcgcc atcgaccgcg 2580

cgggcattgt tggtgctgac ggtcaaaccc atcagggtgc ttttgatctc tcttacctgc 2640

gctgcatacc ggaaatggtc attatgaccc cgagcgatga aaacgaatgt cgccagatgc 2700

tctataccgg ctatcactat aacgatggcc cgtcagcggt gcgctacccg cgtggcaacg 2760

cggtcggcgt ggaactgacg ccgctggaaa aactaccaat tggcaaaggc attgtgaagc 2820

gtcgtggcga gaaactggcg atccttaact ttggtacgct gatgccagaa gcggcgaaag 2880

tcgccgaatc gctgaacgcc acgctggtcg atatgcgttt tgtgaaaccg cttgatgaag 2940

cgttaattct ggaaatggcc gccagccatg aagcgctggt caccgtagaa gaaaacgcca 3000

ttatgggcgg cgcaggcagc ggcgtgaacg aagtgctgat ggcccatcgt aaaccagtac 3060

ccgtgctgaa cattggcctg ccggacttct ttattccgca aggaactcag gaagaaatgc 3120

gcgccgaact cggcctcgat gccgctggta tggaagccaa aatcaaggcc tggctggcat 3180

aaaggaggtt aattggatga agcaactcac cattctgggc tcgaccggct cgattggttg 3240

cagcacgctg gacgtggtgc gccataatcc cgaacacttc cgcgtagttg cgctggtggc 3300

aggcaaaaat gtcactcgca tggtagaaca gtgcctggaa ttctctcccc gctatgccgt 3360

aatggacgat gaagcgagtg cgaaacttct taaaacgatg ctacagcaac agggtagccg 3420

caccgaagtc ttaagtgggc aacaagccgc ttgcgatatg gcagcgcttg aggatgttga 3480

tcaggtgatg gcagccattg ttggcgctgc tgggctgtta cctacgcttg ctgcgatccg 3540

cgcgggtaaa accattttgc tggccaataa agaatcactg gttacctgcg gacgtctgtt 3600

tatggacgcc gtaaagcaga gcaaagcgca attgttaccg gtcgatagcg aacataacgc 3660

catttttcag agtttaccgc aacctatcca gcataatctg ggatacgctg accttgagca 3720

aaatggcgtg gtgtccattt tacttaccgg gtctggtggc cctttccgtg agacgccatt 3780

gcgcgatttg gcaacaatga cgccggatca agcctgccgt catccgaact ggtcgatggg 3840

gcgtaaaatt tctgtcgatt cggctaccat gatgaacaaa ggtctggaat acattgaagc 3900

gcgttggctg tttaacgcca gcgccagcca gatggaagtg ctgattcacc cgcagtcagt 3960

gattcactca atggtgcgct atcaggacgg cagtgttctg gcgcagctgg gggaaccgga 4020

tatgcgtacg ccaattgccc acaccatggc atggccgaat cgcgtgaact ctggcgtgaa 4080

gccgctcgat ttttgcaaac taagtgcgtt gacatttgcc gcaccggatt atgatcgtta 4140

tccatgcctg aaactggcga tggaggcgtt cgaacaaggc caggcagcga cgacagcatt 4200

gaatgccgca aacgaaatca ccgttgctgc ttttcttgcg caacaaatcc gctttacgga 4260

tatcgctgcg ttgaatttat ccgtactgga aaaaatggat atgcgcgaac cacaatgtgt 4320

ggacgatgtg ttatctgttg atgcgaacgc gcgtgaagtc gccagaaaag aggtgatgcg 4380

tctcgcaagc tgaaggaggt taattggatg gcaaccactc atttggatgt ttgcgccgtg 4440

gttccggcgg ccggatttgg ccgtcgaatg caaacggaat gtcctaagca atatctctca 4500

atcggtaatc aaaccattct tgaacactcg gtgcatgcgc tgctggcgca tccccgggtg 4560

aaacgtgtcg tcattgccat aagtcctggc gatagccgtt ttgcacaact tcctctggcg 4620

aatcatccgc aaatcaccgt tgtagatggc ggtgatgagc gtgccgattc cgtgctggca 4680

ggtctgaaag ccgctggcga cgcgcagtgg gtattggtgc atgacgccgc tcgtccttgt 4740

ttgcatcagg atgacctcgc gcgattgttg gcgttgagcg aaaccagccg cacggggggg 4800

atcctcgccg caccagtgcg cgatactatg aaacgtgccg aaccgggcaa aaatgccatt 4860

gctcataccg ttgatcgcaa cggcttatgg cacgcgctga cgccgcaatt tttccctcgt 4920

gagctgttac atgactgtct gacgcgcgct ctaaatgaag gcgcgactat taccgacgaa 4980

gcctcggcgc tggaatattg cggattccat cctcagttgg tcgaaggccg tgcggataac 5040

attaaagtca cgcgcccgga agatttggca ctggccgagt tttacctcac ccgaaccatc 5100

catcaggaga atacataaag gaggttaatt ggatgcggac acagtggccc tctccggcaa 5160

aacttaatct gtttttatac attaccggtc agcgtgcgga tggttaccac acgctgcaaa 5220

cgctgtttca gtttcttgat tacggcgaca ccatcagcat tgagcttcgt gacgatggag 5280

atattcgtct gttaacgccc gttgaaggcg tggaacatga agataacctg atcgttcgcg 5340

cagcgcgatt gttgatgaaa actgcggcag acagcgggcg tcttccgacg ggaagcggtg 5400

cgaatatcag cattgacaag cgtttgccga tgggcggcgg tctcggcggt ggttcatcca 5460

atgccgcgac ggtcctggtg gcattaaatc atctctggca atgcgggcta agcatggatg 5520

agctggcgga aatggggctg acgctgggcg cagatgttcc tgtctttgtt cgggggcatg 5580

ccgcgtttgc cgaaggcgtt ggtgaaatac taacgccggt ggatccgcca gagaagtggt 5640

atctggtggc gcaccctggt gtaagtattc cgactccggt gatttttaaa gatcctgaac 5700

tcccgcgcaa tacgccaaaa aggtcaatag aaacgttgct aaaatgtgaa ttcagcaatg 5760

attgcgaggt tatcgcaaga aaacgttttc gcgaggttga tgcggtgctt tcctggctgt 5820

tagaatacgc cccgtcgcgc ctgactggga caggggcctg tgtctttgct gaatttgata 5880

cagagtctga agcccgccag gtgctagagc aagccccgga atggctcaat ggctttgtgg 5940

cgaaaggcgc taatctttcc ccattgcaca gagccatgct ttaatgtttt ggcggatgag 6000

agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga 6060

atttgcctgg cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga 6120

aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg 6180

catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg 6240

tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag 6300

caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag 6360

cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt 6420

ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 6480

atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 6540

tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 6600

tgaagatcag ttggggcaaa ctattaactg gcgaactact tactctagct tcccggcaac 6660

aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc 6720

cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca 6780

ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga 6840

gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta 6900

agcattggta actgtcagac caagtttact catatatact ttagattgat ttacgcgccc 6960

tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 7020

gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 7080

ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 7140

cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc 7200

tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 7260

ttccaaactt gaacaacact caaccctatc tcgggctatt cttttgattt ataagggatt 7320

ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 7380

tttaacaaaa tattaacgtt tacaatttaa aaggatctag gtgaagatcc tttttgataa 7440

tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 7500

aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 7560

aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 7620

tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc 7680

gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 7740

cctgttacca gtcaggcatt tgagaagcac acggtcacac tgcttccggt agtcaataaa 7800

ccggtaaacc agcaatagac ataagcggct atttaacgac cctgccctga accgacgacc 7860

gggtcgaatt tgctttcgaa tttctgccat tcatccgctt attatcactt attcaggcgt 7920

agcaccaggc gtttaagggc accaataact gccttaaaaa aattacgccc cgccctgcca 7980

ctcatcgcag tactgttgta attcattaag cattctgccg acatggaagc catcacagac 8040

ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg tataatattt 8100

gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact 8160

ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg 8220

gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg 8280

ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa 8340

aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat 8400

acggaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa 8460

cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct gaacggtctg 8520

gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac gatgccattg 8580

ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt ccttagctcc 8640

tgaaaatctc gataactcaa aaaatacgcc cggtagtgat cttatttcat tatggtgaaa 8700

gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg cccagggctt 8760

cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt ccgtcacagg 8820

tatttattcg gcgcaaagtg cgtcgggtga tgctgccaac ttactgattt agtgtatgat 8880

ggtgtttttg aggtgctcca gtggcttctg tttctatcag ctgtccctcc tgttcagcta 8940

ctgacggggt ggtgcgtaac ggcaaaagca ccgccggaca tcagcgctag cggagtgtat 9000

actggcttac tatgttggca ctgatgaggg tgtcagtgaa gtgcttcatg tggcaggaga 9060

aaaaaggctg caccggtgcg tcagcagaat atgtgataca ggatatattc cgcttcctcg 9120

ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc ttacgaacgg 9180

ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg 9240

caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa 9300

atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct 9360

ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc 9420

cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg 9480

gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt 9540

cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga 9600

tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt 9660

tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac 9720

cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag 9780

accaaaacga tctcaagaag atcatcttat taatcagata aaatatttgc tcatgagccc 9840

gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc cagcaaccgc 9900

acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatct gctcatgttt 9960

gacagcttat c 9971

<210> 4

<211> 9034

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 4

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 aggaggttaa ttggatgcga 3240

attggacacg gttttgacgt acatgccttt ggcggtgaag gcccaattat cattggtggc 3300

gtacgcattc cttacgaaaa aggattgctg gcgcattctg atggcgacgt ggcgctccat 3360

gcgttgaccg atgcattgct tggcgcggcg gcgctggggg atatcggcaa gctgttcccg 3420

gataccgatc cggcatttaa aggtgccgat aaccgcgagc tgctacgcga agcctggcgt 3480

cgtattcagg cgaagggtta tacccttggc aacgtcgatg tcactatcat cgctcaggca 3540

ccgaagatgt tgccgcacat tccacaaatg cgcgtgttta ttgccgaaga tctcggctgc 3600

catatggatg atgttaacgt gaaagccact actacggaaa aactgggatt taccggacgt 3660

ggggaaggga ttgcctgtga agcggtggcg ctactcatta aggcaacaaa atgaaggagg 3720

ttaattggat gcataaccag gctccaattc aacgtagaaa atcaacacgt atttacgttg 3780

ggaatgtgcc gattggcgat ggtgctccca tcgccgtaca gtccatgacc aatacgcgta 3840

cgacagacgt cgaagcaacg gtcaatcaaa tcaaggcgct ggaacgcgtt ggcgctgata 3900

tcgtccgtgt atccgtaccg acgatggacg cggcagaagc gttcaaactc atcaaacagc 3960

aggttaacgt gccgctggtg gctgacatcc acttcgacta tcgcattgcg ctgaaagtag 4020

cggaatacgg cgtcgattgt ctgcgtatta accctggcaa tatcggtaat gaagagcgta 4080

ttcgcatggt ggttgactgt gcgcgcgata aaaacattcc gatccgtatt ggcgttaacg 4140

ccggatcgct ggaaaaagat ctgcaagaaa agtatggcga accgacgccg caggcgttgc 4200

tggaatctgc catgcgtcat gttgatcatc tcgatcgcct gaacttcgat cagttcaaag 4260

tcagcgtgaa agcgtctgac gtcttcctcg ctgttgagtc ttatcgtttg ctggcaaaac 4320

agatcgatca gccgttgcat ctggggatca ccgaagccgg tggtgcgcgc agcggggcag 4380

taaaatccgc cattggttta ggtctgctgc tgtctgaagg catcggcgac acgctgcgcg 4440

tatcgctggc ggccgatccg gtcgaagaga tcaaagtcgg tttcgatatt ttgaaatcgc 4500

tgcgtatccg ttcgcgaggg atcaacttca tcgcctgccc gacctgttcg cgtcaggaat 4560

ttgatgttat cggtacggtt aacgcgctgg agcaacgcct ggaagatatc atcactccga 4620

tggacgtttc gattatcggc tgcgtggtga atggcccagg tgaggcgctg gtttctacac 4680

tcggcgtcac cggcggcaac aagaaaagcg gcctctatga agatggcgtg cgcaaagacc 4740

gtctggacaa caacgatatg atcgaccagc tggaagcacg cattcgtgcg aaagccagtc 4800

agctggacga agcgcgtcga attgacgttc agcaggttga aaaataaagg aggttaattg 4860

gatgcagatc ctgttggcca acccgcgtgg tttttgtgcc ggggtagacc gcgctatcag 4920

cattgttgaa aacgcgctgg ccatttacgg cgcaccgata tatgtccgtc acgaagtggt 4980

acataaccgc tatgtggtcg atagcttgcg tgagcgtggg gctatcttta ttgagcagat 5040

tagcgaagta ccggacggcg cgatcctgat tttctccgca cacggtgttt ctcaggcggt 5100

acgtaacgaa gcaaaaagtc gcgatttgac ggcgtttgat gccacctgtc cgctggtgac 5160

caaagtgcat atggaagtcg cccgcgccag tcgccgtggc gaagaatcta ttctcatcgg 5220

tcacgccggg cacccggaag tggaagggac aatgggccag tacagtaacc cggaaggggg 5280

aatgtatctg gtcgaatcgc cggacgatgt gtggaaactg acggtcaaaa acgaagagaa 5340

gctctccttt atgacccaga ccacgctgtc ggtggatgac acgtctgatg tgatcgacgc 5400

gctgcgtaaa cgcttcccga aaattgtcgg tccgcgcaaa gatgacatct gctacgccac 5460

gactaaccgt caggaagcgg tacgcgccct ggcagaacag gcggaagttg tgttggtggt 5520

cggttcgaaa aactcctcca actccaaccg tctggcggag ctggcccagc gtatgggcaa 5580

acgcgcgttt ttgattgacg atgcgaaaga catccaggaa gagtgggtga aagaggttaa 5640

atgcgtcggc gtgactgcgg gcgcatcggc tccggatatt ctggtgcaga atgtggtggc 5700

acgtttgcag cagctgggcg gtggtgaagc cattccgctg gaaggccgtg aagaaaacat 5760

tgttttcgaa gtgccgaaag agctgcgtgt cgatattcgt gaagtcgatt aaatcccggg 5820

aggaggatta ctatatgcaa acggaacacg tcattttatt gaatgcacag ggagttccca 5880

cgggtacgct ggaaaagtat gccgcacaca cggcagacac ccgcttacat ctcgcgttct 5940

ccagttggct gtttaatgcc aaaggacaat tattagttac ccgccgcgca ctgagcaaaa 6000

aagcatggcc tggcgtgtgg actaactcgg tttgtgggca cccacaactg ggagaaagca 6060

acgaagacgc agtgatccgc cgttgccgtt atgagcttgg cgtggaaatt acgcctcctg 6120

aatctatcta tcctgacttt cgctaccgcg ccaccgatcc gagtggcatt gtggaaaatg 6180

aagtgtgtcc ggtatttgcc gcacgcacca ctagtgcgtt acagatcaat gatgatgaag 6240

tgatggatta tcaatggtgt gatttagcag atgtattaca cggtattgat gccacgccgt 6300

gggcgttcag tccgtggatg gtgatgcagg cgacaaatcg cgaagccaga aaacgattat 6360

ctgcatttac ccagcttaaa taactgcagt aaggaggata ccctatggac tttccgcagc 6420

aactcgaagc ctgcgttaag caggccaacc aggcgctgag ccgttttatc gccccactgc 6480

cctttcagaa cactcccgtg gtcgaaacca tgcagtatgg cgcattatta ggtggtaagc 6540

gcctgcgacc tttcctggtt tatgccaccg gtcatatgtt cggcgttagc acaaacacgc 6600

tggacgcacc cgctgccgcc gttgagtgta tccacgctta ctcattaatt catgatgatt 6660

taccggcaat ggatgatgac gatctgcgtc gcggtttgcc aacctgccat gtgaagtttg 6720

gcgaagcaaa cgcgattctc gctggcgacg ctttacaaac gctggcgttc tcgattttaa 6780

gcgatgccga tatgccggaa gtgtcggacc tcgacagaat ttcgatgatt tctgaactgg 6840

cgagcgccag tggtattgcc ggaatgtgcg gtggtcaggc attagattta gacgcggaag 6900

gcaaacacgt acctctggac gcgcttgagc gtattcatcg tcataaaacc ggcgcattga 6960

ttcgcgccgc cgttcgcctt ggtgcattaa gcgccggaga taaaggacgc cgtgctctgc 7020

cggtactcga caagtatgca gagagcatcg gccttgcctt ccaggttcag gacgacatcc 7080

tggatgtggt gggagatact gcaacgttgg gaaaacgcca gggtgccgac cagcaacttg 7140

gtaaaagtac ctaccctgca cttctgggtc ttgagcaagc ccggaagaaa gcccgggatc 7200

tgatcgacga tgcccgtcag tcgctgaaac aactggctga acagtcactc gatacctcgg 7260

cactggaagc gctagcggac tacatcatcc agcgtaataa ataacaccgc ggtggcggcc 7320

gctctagaac tagtggatcc cccgggctgc aggaattcga tatcaagctt atcgataccg 7380

tcgacctcga gggggggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg 7440

cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 7500

ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 7560

taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 7620

cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgcatg 7680

cataaaaact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca 7740

tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc 7800

attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa 7860

aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat 7920

tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc 7980

agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga 8040

gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg 8100

cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc 8160

agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag 8220

taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc 8280

tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg 8340

taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg 8400

acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac 8460

ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac 8520

cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg 8580

agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg 8640

tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg 8700

agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac 8760

tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg 8820

ataatctcat gaccaaaatc caggtggcac ttttcgggga aatgtgcgcg cccgcgttcc 8880

tgctggcgct gggcctgttt ctggcgctgg acttcccgct gttccgtcag cagcttttcg 8940

cccacggcct tgatgatcgc ggcggccttg gcctgcatat cccgattcaa cggccccagg 9000

gcgtccagaa cgggcttcag gcgctcccga aggt 9034

<210> 5

<211> 6915

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 5

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

<210> 6

<211> 11133

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 6

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> 7

<211> 10396

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 7

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> 8

<211> 8096

<212> DNA

<213> Artificial Sequence (Artificial Sequence)

<400> 8

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 taaaggagga attaaccatg aaacatctgc atcgattctt tagcagtgat 6840

gcctcgggag gcattattct tatcattgcc gctatcctgg cgatgattat ggccaacagc 6900

ggcgcaacca gtggatggta tcacgacttt ctggagacgc cggttcagct ccgggttggt 6960

tcactcgaaa tcaacaaaaa catgctgtta tggataaatg acgcgctgat ggcggtattt 7020

ttcctgttag tcggtctgga agttaaacgt gaactgatgc aaggatcgct agccagctta 7080

cgccaggccg catttccagt tatcgccgct attggtggga tgattgtgcc ggcattactc 7140

tatctggctt ttaactatgc cgatccgatt acccgcgaag ggtgggcgat cccggcggct 7200

actgacattg cttttgcact tggtgtactg gcgctgttgg gaagtcgtgt tccgttagcg 7260

ctgaagatct ttttgatggc tctggctatt atcgacgatc ttggggccat cattatcatc 7320

gcattgttct acactaatga cttatcgatg gcctctcttg gcgtcgcggc tgtagcaatt 7380

gcggtactcg cggtattgaa tctgtgtggt gcacgccgca cgggcgtcta tattcttgtt 7440

ggcgtggtgt tgtggactgc ggtgttgaaa tcgggggttc acgcaactct ggcgggggta 7500

attgtcggct tctttattcc tttgaaagag aagcatgggc gttctccagc gaagcgactg 7560

gagcatgtgt tgcacccgtg ggtggcgtat ctgattttgc cgctgtttgc atttgctaat 7620

gctggcgttt cactgcaagg cgtcacgctg gatggcttga cctccattct gccattgggg 7680

atcatcgctg gcttgctgat tggcaaaccg ctggggatta gtctgttctg ctggttggcg 7740

ctgcgtttga aactggcgca tctgcctgag ggaacgactt atcagcaaat tatggtggtg 7800

gggatcctgt gcggtatcgg ttttactatg tctatcttta ttgccagcct ggcctttggt 7860

agcgtagatc cagaactgat taactgggcg aaactcggta tcctggtcgg ttctatctct 7920

tcggcggtaa ttggatacag ctggttacgc gttcgtttgc gtccatcagt ttgacaaagc 7980

ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg 8040

ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggat 8096

Claims (10)

1. The application of nucleic acid for encoding the NhaA protein in preparing the engineering bacteria for producing the albizium julibrissin, wherein the amino acid sequence of the NhaA protein is the amino acid sequence shown in SEQ ID NO. 1; the engineering bacteria is escherichia coli.

2. The use according to claim 1, wherein the nucleic acid encoding a NhaA protein is a nucleic acid consisting of the nucleotide sequence shown in SEQ ID No. 2; the engineering bacteria for producing the farnesene are engineering bacteria capable of producing the farnesene by fermenting cellulose hydrolysate.

3. The use according to claim 2, wherein the engineered bacterium capable of producing farnesene by fermentation using a cellulose hydrolysate can have the expression of isopentenyl pyrophosphate isomerase, farnesene pyrophosphate synthase and farnesene synthase, and can have the expression of a plurality of enzymes constituting a mevalonate pathway or a plurality of enzymes constituting a deoxyxylulose-5-phosphate pathway;

the plurality of enzymes constituting the mevalonate pathway include acetoacetyl-CoA thiolase, HMG-CoA synthase, HMG-CoA reductase, mevalonate kinase, phosphomevalonate kinase, and mevalonate pyrophosphate decarboxylase; the acetoacetyl CoA thiolase is an atoB enzyme and/or an MvaE enzyme; the HMG-CoA synthase is HMGS enzyme and/or MvaS enzyme, and the HMG-CoA reductase is HMGR enzyme and/or MvaE enzyme;

the various enzymes constituting the deoxyxylulose-5-phosphate pathway include 1-deoxy-D-xylulose-5-phosphate synthase, 1-deoxy-D-xylulose-5-phosphate reductoisomerase, cytidine-2C-methyl-D-erythritol 4-diphosphate synthase, 2C-methyl-D-erythrose-2, 4-cyclic diphosphate synthase, 1-hydroxy-2-methyl-2- (E) -butenyl 4-diphosphate synthase, and isopentenyl/dimethylallyl diphosphate synthase.

4. A method for preparing an engineering bacterium for producing silktree albizzia, which is characterized by comprising the following steps: the transformed host bacteria of the engineering bacteria can have the expression of isopentenyl pyrophosphate isomerase, farnesene pyrophosphate synthetase and farnesene synthetase, the expression of a plurality of enzymes forming a mevalonate pathway or a plurality of enzymes forming a deoxyxylulose-5-phosphate pathway, and the over-expression of NhaA protein by transforming the host bacteria of the engineering bacteria into recombinant plasmids or transforming chromosome integration knock-in;

the amino acid sequence of the NhaA protein is an amino acid sequence shown in SEQ ID NO. 1;

the engineering bacteria is escherichia coli.

5. The method according to claim 4, wherein the engineered bacteria host bacteria can have the overexpression of citrate lyase and/or acetate kinase by transforming the engineered bacteria host bacteria with recombinant plasmid transfer or chromosome integration knock-in.

6. The method of claim 4 or 5, wherein the plurality of enzymes that make up the mevalonate pathway comprise acetoacetyl-CoA thiolase, HMG-CoA synthase, HMG-CoA reductase, mevalonate kinase, phosphomevalonate kinase, and mevalonate pyrophosphate decarboxylase; the acetoacetyl CoA thiolase is an atoB enzyme and/or an MvaE enzyme; the HMG-CoA synthase is HMGS enzyme and/or MvaS enzyme, and the HMG-CoA reductase is HMGR enzyme and/or MvaE enzyme;

the various enzymes constituting the deoxyxylulose-5-phosphate pathway include 1-deoxy-D-xylulose-5-phosphate synthase, 1-deoxy-D-xylulose-5-phosphate reductoisomerase, cytidine-2C-methyl-D-erythritol 4-diphosphate synthase, 2C-methyl-D-erythrose-2, 4-cyclic diphosphate synthase, 1-hydroxy-2-methyl-2- (E) -butenyl 4-diphosphate synthase, and isopentenyl/dimethylallyl diphosphate synthase.

7. An engineered bacterium produced by the method of any one of claims 4 to 6.

8. 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 bacterium of claim 7 into a liquid fermentation raw material containing the hydrolysate containing monosaccharide, and performing fermentation culture to obtain a cultured material.

9. The method of claim 8, wherein the cellulose-containing feedstock comprises at least one of straw, bark, wood chips, corn cobs, rice bran, and wheat bran;

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.

10. The method of claim 8 or 9, 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 is (10-300) x 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.