KR20120006463A - Agrobacterium tumefaciens species capable of reconstituting infection activity of cucumber mosaic virus in plant - Google Patents

Abstract

PURPOSE: An Agrobacterium tumefaciens mixture which is able to reproduce CMV(cucumber mosaic virus) infection is provided to identify plant development and differentiation. CONSTITUTION: An Agrobacterium tumefaciens mixture contains three kinds of Agrobacterium tumefaciens of sequence numbers 1-3. The Agrobacterium tumefaciens is deposited by KACC95081P, KACC95082P, or KACC95083P. The plant is Nicotiana benthamiana.

Description

Agrobacterium tumefaciens species capable of reconstituting infection activity of Cucumber mosaic virus in plant

The present invention relates to Agrobacterium tumefacience capable of reproducing the infectious activity of Cucumber mosaic virus (CMV) in plants.

Numerous technologies are required for biotechnology research, and among them, transformation technology is one of the important technologies that are essentially indispensable. Transformation literally means introducing a trait (gene) that it does not have or removing a trait that it has. Through this, the function of the trait can be analyzed.

In particular, plant transformation technology is an important technology that can be very usefully used for analyzing the function or expression of plant genes. It identifies various phenomena that are involved in the development and differentiation of plants, or develops new functional varieties through the introduction of useful genes. It has been used as a technology. In general, plant transformation has been developed based on tissue culture technology, but when using tissue culture, there is a disadvantage that requires a lot of time and effort due to a long regeneration period. Serious problems, such as the appearance of, have been raised. Therefore, a plant transformation technology that does not undergo tissue culture has been developed, which transforms the cells located at the growth point site as the plant grows without undergoing tissue culture, and then transforms the stem redifferentiating from the transformed cells. It is a technology that can obtain a genetically stable transgenic plant by obtaining the seed. Derived from this technology is the Agro-infiltration method using the Agrobacterium strain.

Agrobacterium tumefaciens tumefaciens ) has long been known as a soil bacterium that causes Crown gall disease in many dicotyledonous plants. In the 1970s, it was found that Agrobacterium's Ti plasmid was associated with pathogenicity, and that T-DNA, part of the Ti plasmid, was integrated into the plant genome. Later, it was found that T-DNA contains hormone-synthesizing genes (cytokinin and auxin) necessary for the formation of myohead carcinoma, and these genes are derived from bacteria, but expressed in plants. A group of genes located in the virulence region (Vir region) of the Ti plasmid are required for the resection of T-DNA and their migration to plants, and also the right border sequence and the left side located at the opposite ends of the T-DNA. It has also been found that border sequences, termed border sequences, are required for the ablation.

Cucumber mosaic virus ( CMV ) was first reported in 1916 (Doolittle, S, The mosaic diseases of cucurbits, USDA Agri. Bull (1961) 879:69). CMV is a multiparticulate virus with a ball-shaped three-segment genome with a diameter of 28-30 nm. It exhibits mosaic symptoms on leaves in cucumbers (Cucumis sativus), etc., and the fruit becomes smaller and deformed, leading to a significant drop in yield as well as product value. CMV has a very wide host range and is distributed throughout the world.

The CMV genome consists of three positive-sense single-stranded RNAs (RNA1, RNA2 and RNA3) and RNA4 derived from RNA3. RNA1 and RNA2 are pathogenic and encode proteins 1a and 2a, respectively, involved in viral replication. RNA3 encodes 3a protein and coat protein ( CP ). The 3a protein is involved in cell-to-cell migration functions. The coat protein is translated from subgenome RNA (RNA4) transcribed from RNA3. These characteristics also appear in bromovirus, ilarvirus and alfalfa mosaic virus.

International Patent Application No. PCT/JP1999/005386 (International Application Date 1999. 09. 30) relates to a plant transformation vector. Vectors for plant transformation that use bacteria and thus lower the likelihood of transfer of DNAs other than T-DNA into plant chromosomes, in particular, these vectors are right and left border sequences that can be recognized by Agrobacterial vir proteins. Field, a T-DNA sequence that is located between the border sequences and into which a gene to be introduced into a plant can be inserted, and a replication initiation point that enables the replication of the vectors in bacteria, and a plurality of left borders Vectors used for plant transformation, characterized by having sequences, are disclosed.

Therefore, the present inventors put infectious clones of cucumber mosaic virus isolated from red beans in domestic packaging in a binary vector and introduce them into plants using Agrobacterium tumefaciens, agro-infiltration ( Agro-infiltration) method was used to introduce viral genes into plants to confirm that infectious CMV activity was reconstituted, and the present invention was completed.

An object of the present invention is to provide Agrobacterium tumefacience capable of reproducing the infectious activity of Cucumber mosaic virus (CMV) in plants.

In order to achieve the above object, the present invention is Agrobacterium tumefaciens each transformed with SEQ ID NO: 1 to SEQ ID NO: 3 capable of reproducing the infectious activity of Cucumber mosaic virus (CMV) in plants ( Agrobacterium tumefaciens ) It provides a mixture of Agrobacterium tumefaciens containing three species. In addition, Agrobacterium Tumefacience of the accession number KACC95081P, KACC95082P or KACC95083P is provided. Preferably, the plant is a mixture of Agrobacterium tumefaciens, characterized in that Nicotiana benthamiana (Nicotiana benthamiana).

The present invention relates to an Agrobacterium tumefaciens mixture capable of reproducing the infectious activity of Cucumber mosaic virus (CMV) in plants. Plant transformation technology using traditional tissue culture has a disadvantage that requires a lot of time and effort due to a long regeneration period as well as low efficiency of the transformation process, and somatic cell mutation or natural malformation occurs during the regeneration process. There was a serious problem. The use of the Agrobacterium tumefaciens mixture of the present invention has the advantage of being quick and convenient when compared to traditional plant transformation in introducing genes into plants. The development of gene carriers for plant transformation with high utilization can be used as a technology to identify various phenomena involved in the development and differentiation of plants, or to develop new functional varieties through the introduction of useful genes.

1 shows a schematic diagram of the structure of a pSNU1 vector.
Multiple cloning site part: 1-68 bp, cis- cleaving ribozyme sequence (Rz): 69-119 bp, NOS poly-A: 184-436 bp, T-border (right): 474-499 bp, pVS1 sta :1540-2540bp, pVS1 rep:3223-4223bp, pBR322 bom:4543-4803bp, pBR322 ori:4943-5223bp, Kanamycin resistance gene (aadA):5514-6308bp, T-border (left):6733-6758bp , Cauliflower mosaic virus ( CaMV ) 35S duplication promoter: 6877-7628bp
Figure 2 shows a schematic diagram of the structure of an infectious clone of Cucumber mosaic virus ( CMV). A) a schematic diagram of pCkpaR1 from which CMV RNA1 is cloned, B) pCkpaR2 from which CMV RNA2 is cloned, C) pCkpaR3 from which CMV RNA3 is cloned; CP: coat protein.
Figure 3 shows a schematic diagram of the inoculation method of infectious clones through agro-infiltration (agro-infiltration).
Figure 4 is Nicotiana Ventamiana (Nicotiana) infected with CMV benthamiana ) It shows the symptom expression picture of the plant.
5 is a picture of a gel confirming CMV infectivity by using the RT-PCR technique.
Lane 1: size marker (size marker), lane 2: control (mock), lane 3: agro-infiltration (agro-infiltration) via a CMV infectious clone was inoculated experimental group (upper lobe), lane 4: agro- Experimental sphere inoculated with CMV infectious clone through agro-infiltration (inoculated leaves)

Embodiment of the first aspect of the present invention is cucumber mosaic virus; the respective transformants to be reproduced to an active infection in plants SEQ ID NO: 1 to SEQ ID NO: 3 in the (Cucumber mosaic virus CMV) Agrobacterium-to mepa sieon switch (Agrobacterium tumefaciens ) It provides a mixture of Agrobacterium tumefaciens containing three species. Preferably, the plant is a mixture of Agrobacterium tumefaciens, characterized in that Nicotiana benthamiana (Nicotiana benthamiana).

A second aspect of the present invention also provides Agrobacterium tumefaciens with accession numbers KACC95081P, KACC95082P or KACC95083P.

The strains containing the genes pCkpaR1 (SEQ ID NO: 1), pCkpaR2 (SEQ ID NO: 2), and pCkpaR3 (SEQ ID NO: 3) were deposited with the Institute of Agriculture and Biotechnology on May 21, 2008 under the deposit numbers KACC 95081P, KACC 95082P, and KACC 95083P, respectively. .

The term'Agro-infiltration' used in the present invention refers to a method for introducing and expressing a gene into a plant or producing a desired protein in a plant. This is a method of injecting a suspension of Agrobacterium containing the gene to be transferred into the plant leaf by applying pressure using a needleless syringe to transfer the desired gene to the plant cell. This has the advantage of being quick and convenient compared to traditional plant transformation.

The term'vector' used in the present invention refers to a nucleic acid molecule capable of transporting by linking another nucleic acid to it. A preferred vector is a vector capable of self-replicating and expressing the nucleic acid to which it is bound.

The term'transformation' used in the present invention refers to a change in the genotype of a cell by absorption of foreign DNA or RNA into a cell. Host cells include, but are not limited to, plant cells, prokaryotic cells, yeast cells, and insect cells.

Hereinafter, the present invention will be described in more detail by examples.

However, the following examples are merely illustrative of the present invention, and the contents of the present invention are not limited to the following examples.

< Example 1> Design and manufacture of a recombinant vector for plant transformation using an infectious clone of cucumber mosaic virus

The pSNU1 vector was used as a vector for carrying the infectious clone of Cucumber mosaic virus ( CMV ) (FIG. 1 ). pSNU1 vector is the left border of T-DNA, double 35S promoter of Cauliflower mosaic virus (CaMV ), multiple cloning site ( MCS ; Eco RI , Xba I , Stu I , Nco) I , Bam HI , Kpn I , Sac I , Mlu I , Xho I , Srf I , Sma I , Spe I), cis -cleaving ribozyme sequence(Rz), NOS terminator(NOS poly-A), right side of T-DNA It contains the right border in order. Therefore, it can be transformed into Agrobacterium to deliver a desired gene to a plant through Agro-infiltration. In addition, the pSNU1 vector contains a kanamycin resistance gene.

In order to produce a CMV infectious clone capable of agro-infiltration based on T-DNA, a CMV isolate isolated from red beans in domestic packaging was used. Total RNA was extracted from red beans infected with the CMV isolate. From this total RNA, using a primer (5'-G GAATTC TGGTCTCCTTTTRGAGRCC-3' (SEQ ID NO: 4) or 5'-AG GGATCC TGGTCTCCTTTTRGAGRCC-3' (SEQ ID NO: 5)) through reverse transcription reaction to CMV RNA1, RNA2, RNA3 For cDNA was synthesized. To amplify the CMV RNA1 from the cDNA synthesis primer (5'-A GGATCC GTTTATTTACAAGAGCGTACGG-3 '( SEQ ID NO: 6) and 5'-GGATCC TGGTCTCCTTTTRGAGRCC AG-3' (SEQ ID NO: 7)) was subjected to PCR reaction using a . In order to amplify the CMV RNA2 primer (5'-GGATCC GTTTATTTACAAGAGCGTACGG AG-3 '(SEQ ID NO: 8) and 5'-G ACTAGT GGTCTCCTTTTRGAGRCC-3' ( SEQ ID NO: 9)) to, in order to amplify a CMV RNA3 primer (5 ' the -AG GGATCC GTAATCTTACCACTGTG-3 '(SEQ ID NO: 10) and 5'-GGATCC TGGTCTCCTTTTRGAGRCC AG-3' (SEQ ID NO: 11)) was used. After denaturing at 94°C for 3 minutes using each primer, repeat 35 cycles of 30 seconds at 94°C, 30 seconds at 55°C, 3 minutes at 72°C, and react at 72°C for 10 minutes. DNA fragments containing the nucleotide sequence of were amplified.

In order to clone into the pSNU1 vector, the CMV RNA1 PCR product was treated with BamH I and cloned into the corresponding position of the vector. For CMV RNA2, Bam HI and Spe I, and for CMV RNA3, Bam HI restriction enzyme was used. The resulting CMV RNA1 cloned vector was named pCkpaR1 (SEQ ID NO: 1), the CMV RNA2 cloned vector was named pCkpaR2 (SEQ ID NO: 2), and the CMV RNA3 cloned vector was named pCkpaR3 (SEQ ID NO: 3) ( Fig. 2 ).

Strains containing the pCkpaR1 (SEQ ID NO: 1), pCkpaR2 (SEQ ID NO: 2), and pCkpaR3 (SEQ ID NO: 3) genes were deposited with the Agricultural Biotechnology Research Institute on May 21, 2008 under the deposit numbers KACC 95081P, KACC 95082P, and KACC 95083P, respectively. Became.

<Example 2> Agrobacterium-bit-fill migration check inoculation and infectivity of the infectious clone with (Agro-infiltration)

Using the plasmid DNA of each clone, Agrobacterium strain GV2260 was transformed through an electroporation method. Agrobacterium transformed with pCkpaR1, pCkpaR2, and pCkpaR3, respectively, in 5 ml of YEP liquid medium (including 50 mg/ml of kanamycin and 50 mg/ml of rifampicin) at 30° C. for 16 hours After shaking culture during the period, 1 ml of the primary culture solution was inoculated into 50 ml of new YEP medium (including 50 mg/ml of kanamycin, 50 mg/ml of rifampicin and 20 μM of acetosyringon) at 30°C. Shake culture was carried out for 6 hours under conditions. The culture solution was centrifuged at 4800G for 10 minutes to precipitate Agrobacterium, and in MMA infiltration buffer (MS salts, 10mM MES, pH5.6, 200uM acetosyringon) at a wavelength of 600nm. It was re-suspended to a concentration of OD 1. Then, the suspension was cultured with shaking for 2 hours at 30°C. After mixing the Agrobacterium cultures transformed with pCkpaR1, pCkpaR2 and pCkpaR3 in the same ratio, Nicotiana ventamiana (Nicotiana benthamiana ) on the hypocotyl surface of the leaf using a 1ml syringe was inflation (infiltration) under pressure. 5-8 days after Agro-infiltration, the infectivity of the CMV DNA clone was confirmed through RT-PCR and observation of the development of symptoms ( FIG. 3 ). As a result, CMV infectious clones were collected from Nicotiana ventamiana through Agro-infiltration. benthamiana ) 7 days after inoculation on the plant, the symptoms indicated by CMV could be observed from the upper leaf ( FIG. 4 ).

In addition, CMV infectious clones were collected from Nicotiana ventamiana through Agro-infiltration. benthamiana ) 7 days after inoculation to plants, after extracting total RNA from the upper leaves, primers specific for CMV RNA3 (5'-TCCCTGTTGAGCCCCCTTACTTT-3' (SEQ ID NO: 12) and 5'-CAACTCAGATCCCGCCACAGA-3' (SEQ ID NO: 13)) PCR product was confirmed through gel electrophoresis by performing RT-PCR using. PCR conditions were denatured at 94°C for 2 minutes, then repeated 35 cycles at 94°C for 20 seconds, 60°C for 30 seconds, and 1 minute at 72°C for 1 cycle, and then reacted at 72°C for 10 minutes.

As a result of the confirmation, it was possible to confirm the PCR product of the expected size for the plant inoculated with the CMV infectious clone, and thus it was confirmed that the plant became a systemic infection by CMV ( FIG. 5 ).

Institute of Agricultural Biotechnology KACC95081 20080414 Institute of Agricultural Biotechnology KACC95082 20080414 Institute of Agricultural Biotechnology KACC95083 20080414

<110> Seoul National University Industry Foundation Rural Development Administration <120> The mixture composition comprising Agrobacterium tumefaciens species capable of reconstituting infection activity of Cucumber mosaic virus in plant <160> 13 <170> KopatentIn 1.71 <210> 1 <211> 10993 <212> DNA <213> Artificial Sequence <220> <223> pCkpaR1 <400> 1 gaattctcta gaaggcctcc atggggatcc gtttatttac aagagcgtac ggttcaatcc 60 ctgcctcctc tgtaaaatta ccctttgaaa acctctcttt tctaatcttt tcttagtaat 120 tcctatggcg acgtcctcgt tcagtatcaa tgaattggta gcctcccacg gcgataaagg 180 actactcgcg accgccctcg ttgataaaac agctcatgag cagctcgagg agcaattaca 240 acatcaacgt agaggccgta aggtctacat tcggaacgtt ctgagtgtaa aggattccga 300 agtcatccgg aatcggtatg gaggaaagta cgacctccat cttacccagc aggagtttgc 360 tccccacggc ctagctggtg ccctccgctt gtgtgaaact ctcgattgtc tagactcttt 420 cccttcatcg ggtctgcggc aggacctcgt cttagacttc ggaggaagtt gggtcacaca 480 ttacctccgc ggacacaacg tacactgctg ttccccttgt ttgggtatcc gcgataagat 540 gcgccacgcg gaacgtttaa tgaacatgcg caagatcatc ttgaacgatc cacaacagtt 600 cgatggtcga cagccggatt tttgcactca tccagctgct gattgcaaag tacaagctca 660 ctttgctata tctattcatg gaggttatga tatgggcttt agaggattat gtgaggcgat 720 gaacgctcac ggaaccacta ttttgaaggg aacgatgatg ttcgatggtg cgatgatgtt 780 tgacgaccaa ggcacgatac ccgaacttaa ttgccagtgg agaaaaatta ggagcgcatt 840 ctctgaagct gaagacgtca catctctgtc tggtaaaatt gattcctctg gattcaccgg 900 tgtgcgtaag ttcaagactt tggttgcttt tgattttatc aatgagtcta ccatgtcgta 960 cgttcatgat tggaagaata taaaatcctt tctaacggac cagacttact cgtataaagg 1020 gatgacttat ggcattgaac gatgtgtcat taatgcgggt atcatgacgt ataagataat 1080 cggtgtgcct gggatgtgtc cgcccgaact cattcgacat tgtatttggt tcccctctat 1140 taaagattat gttggtttaa agattcccgc gtcgcaggac ttggttaagt ggaaaacagt 1200 gcgtgtttta atgtcaacat tacgtgaacc agaacagatt gctatgaggt gttacaacga 1260 taagaaagcg tggatggaac aattcaaggt tatcctgggt gttttgtctg cgaaatcatc 1320 taccattgtc attaatggta tgtccatgca atctggtgag cgtatagata ttaatgatta 1380 ccattatatt ggtttcgcta ttcttctgca cacaaaaatg aagtatgaac aactgggaaa 1440 aatgtatgat atgtggaatg cttcgagcat atcgaagtgg tttgcagcgt tgactcgacc 1500 gctgcgtgtg tttttctcca gtgttgttca cgcactgttc ccgactttga gaccccgcga 1560 ggaaaaagaa ttcctgatta agctctccac cttcgttact tttaatgaag agtgctcatt 1620 tgatggtgga gaggaatggg acgttatatc atctgctgca tatgttgcta cgcaggctgt 1680 taccgatggg aagattttgg ctacgcagaa agctgagaag cttgctgaga agcttgcaca 1740 acccgtgatt gaggtatcgg acagacctga ggtgtcatct ccaacacctg atgatacagc 1800 tgatgtttgt ggaaaagagc aagaagtttc ggaacttgac tctttatcag ctcagacacg 1860 ttcccccatc accagaattg ctgaaagggc tactgctatg ttagagtatg ccgcttatga 1920 gaaacagtta cacgacacca cagtgtctaa tctgaaacgt atttggaaca tggcgggcgg 1980 tgatgataag agaaattccc ttgagggtaa tctgaagttc gttttcgata cgtatttcac 2040 tgttgatccc atggtgaata ttcatttctc cacgggtcgg tggatgcgtc ctgtgcccga 2100 agggattatc tattctgttg gttataatga acgcggttta ggtccgaagt cggatggaga 2160 gctttacatt gtcaacagag agtgcgtgat ttgcaatagc gagtctttgt ctgctatcac 2220 tcgttctctt caggccccga ccggaaccat tagtcaagtc gatggggttg ccggttgcgg 2280 gaagacaacg gcaataaaat ccatttttga gccatccact gacatgatcg ttaccgcgaa 2340 caagaaatcc gcccaagatg ttcgtatggc actgtttaaa tcatcggatt ccaaagaagc 2400 ttgcaccttt gttcgtaccg ccgattctgt cctacttaat gaatgtccga ctgttagtag 2460 ggttttaatt gatgaggttg tgctgcttca ctttggtcaa ttgtgtgccg tgatgtctaa 2520 gctgaaggct gtgcgggcta tatgtttcgg ggattcggag caaattgcct tttcttctcg 2580 cgacgcctcg tttgacatgc gtttccctaa gattattcct gatgaaacta gtgatgcgga 2640 tactacattc cgtagcccac aagacgttgt gccgcttgtg cgtttaatgg ctacgaaggc 2700 tcttccgaaa ggaactcatt cgaaatacac gaaatgggtt tctcaatcta aagtgaaaag 2760 atctattaca tctcgtgcca tcgctagcgt gacattggtt gatctggatt cttccaggtt 2820 ttacattacg atgacccaag ctgataaggc ctcactgatt tcaagagcga aagagatgaa 2880 tctaccaaag actttctggg acgaaaggat taaaactgtg catgagtctc aaggcatttc 2940 tgaagatcat gttactttgg taagattaaa gagcacaaag tgtgacctgt tcaagaaatt 3000 ttcatattgt cttgttgctt tgacgagaca caaggtcaca ttccgctacg agtactgcgg 3060 tgtactgaac ggcgatttaa tcgccgaatg tattgctcgt gcttagcgga ctccctcctt 3120 gtgaggcggg aggtgagttg gcagtattgc tataaactgt ctgaagtcac taaatacatt 3180 tgtagtgaac gggttgtcca tccagcttac ggctaaaatg gtcagttgta gaaaaatcta 3240 cgccggaggc ttacaagtct ccgaggcgcc tttgaaacca tctcctaggt ttcttcggaa 3300 ggacttcggt ccgtgtactt ctagcacaag atgctagttt cagggtacag gtgccccccc 3360 cgcttggtgg ggcctctaaa aggagaccag gatccggtac cgagctcacg cgtctcgagg 3420 cccgggcact agtgtctgta cttatatcag tacactgacg agtccctaaa ggacgaaacg 3480 ggagaacgct agccaccacc accaccacca cgtgtgaatt acaggtgacc agctcgaatt 3540 tccccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc 3600 ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt 3660 aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt 3720 aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt 3780 catctatgtt actagatcgg gaattaaact atcagtgttt gacaggatat attggcgggt 3840 aaacctaaga gaaaagagcg tttattagaa taacggatat ttaaaagggc gtgaaaaggt 3900 ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggatcaaagt 3960 actttgatcc aacccctccg ctgctatagt gcagtcggct tctgacgttc agtgcagccg 4020 tcttctgaaa acgacatgtc gcacaagtcc taagttacgc gacaggctgc cgccctgccc 4080 ttttcctggc gttttcttgt cgcgtgtttt agtcgcataa agtagaatac ttgcgactag 4140 aaccggagac attacgccat gaacaagagc gccgccgctg gcctgctggg ctatgcccgc 4200 gtcagcaccg acgaccagga cttgaccaac caacgggccg aactgcacgc ggccggctgc 4260 accaagctgt tttccgagaa gatcaccggc accaggcgcg accgcccgga gctggccagg 4320 atgcttgacc acctacgccc tggcgacgtt gtgacagtga ccaggctaga ccgcctggcc 4380 cgcagcaccc gcgacctact ggacattgcc gagcgcatcc aggaggccgg cgcgggcctg 4440 cgtagcctgg cagagccgtg ggccgacacc accacgccgg ccggccgcat ggtgttgacc 4500 gtgttcgccg gcattgccga gttcgagcgt tccctaatca tcgaccgcac ccggagcggg 4560 cgcgaggccg ccaaggcccg aggcgtgaag tttggccccc gccctaccct caccccggca 4620 cagatcgcgc acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa agaggcggct 4680 gcactgcttg gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag cgaggaagtg 4740 acgcccaccg aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac cgaggccgac 4800 gccctggcgg ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg caccaggacg 4860 gccaggacga accgtttttc attaccgaag agatcgaggc ggagatgatc gcggccgggt 4920 acgtgttcga gccgcccgcg cacgtctcaa ccgtgcggct gcatgaaatc ctggccggtt 4980 tgtctgatgc caagctggcg gcctggccgg ccagcttggc cgctgaagaa accgagcgcc 5040 gccgtctaaa aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt 5100 atatgatgcg atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt 5160 acttaaccag aaaggcgggt caggcaagac gaccatcgca acccatctag cccgcgccct 5220 gcaactcgcc ggggccgatg ttctgttagt cgattccgat ccccagggca gtgcccgcga 5280 ttgggcggcc gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat 5340 tgaccgcgac gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg gagcgcccca 5400 ggcggcggac ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca 5460 gccaagccct tacgacatat gggccaccgc cgacctggtg gagctggtta agcagcgcat 5520 tgaggtcacg gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac 5580 gcgcatcggc ggtgaggttg ccgaggcgct ggccgggtac gagctgccca ttcttgagtc 5640 ccgtatcacg cagcgcgtga gctacccagg cactgccgcc gccggcacaa ccgttcttga 5700 atcagaaccc gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc 5760 aaaactcatt tgagttaatg aggtaaagag aaaatgagca aaagcacaaa cacgctaagt 5820 gccggccgtc cgagcgcacg cagcagcaag gctgcaacgt tggccagcct ggcagacacg 5880 ccagccatga agcgggtcaa ctttcagttg ccggcggagg atcacaccaa gctgaagatg 5940 tacgcggtac gccaaggcaa gaccattacc gagctgctat ctgaatacat cgcgcagcta 6000 ccagagtaaa tgagcaaatg aataaatgag tagatgaatt ttagcggcta aaggaggcgg 6060 catggaaaat caagaacaac caggcaccga cgccgtggaa tgccccatgt gtggaggaac 6120 gggcggttgg ccaggcgtaa gcggctgggt tgtctgccgg ccctgcaatg gcactggaac 6180 ccccaagccc gaggaatcgg cgtgacggtc gcaaaccatc cggcccggta caaatcggcg 6240 cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac 6300 gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca 6360 aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg 6420 acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca 6480 gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga 6540 tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca 6600 gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc 6660 gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg 6720 tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct 6780 gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc 6840 gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg 6900 aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca 6960 cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg 7020 gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat 7080 ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt 7140 tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg 7200 cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat 7260 ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag 7320 gtcgaaaagg tctctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg 7380 ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt 7440 aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta 7500 ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag 7560 agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc 7620 ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag 7680 ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc 7740 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc 7800 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt 7860 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact 7920 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa 7980 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca 8040 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg 8100 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc 8160 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc 8220 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac 8280 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc 8340 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata 8400 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc 8460 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca 8520 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag 8580 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta 8640 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg 8700 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc 8760 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt 8820 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat tctaggtact 8880 aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca 8940 gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg atcgaccgga 9000 cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc 9060 cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga 9120 caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa 9180 atggagtgtc ttcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt 9240 aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga 9300 tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt 9360 catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc 9420 agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata 9480 gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca 9540 tttttaaata taggttttca ttttctccca ccagcttata taccttagca ggagacattc 9600 cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc 9660 tcattttagc catttattat ttccttcctc ttttctacag tatttaaaga taccccaaga 9720 agctaattat aacaagacga actccaattc actgttcctt gcattctaaa accttaaata 9780 ccagaaaaca gctttttcaa agttgttttc aaagttggcg tataacatag tatcgacgga 9840 gccgattttg aaaccgcggt gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca 9900 tgctaccctc cgcgagatca tccgtgtttc aaacccggca gcttagttgc cgttcttccg 9960 aatagcatcg gtaacatgag caaagtctgc cgccttacaa cggctctccc gctgacgccg 10020 tcccggactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg 10080 agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac gcttagacaa 10140 cttaataaca cattgcggac gtttttaatg tactgaatta acgccgaatt aattcctagg 10200 ccaccatgtt gggcccggcg cgccaagctt gcatgcctgc aggtcaacat ggtggagcac 10260 gacactctcg tctactccaa gaatatcaaa gatacagtct cagaagacca gagggctatt 10320 gagacttttc aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc 10380 tgtcacttca tcgaaaggac agtagaaaag gaagatggct tctacaaatg ccatcattgc 10440 gataaaggaa aggctatcgt tcaagatgcc tctaccgaca gtggtcccaa agatggaccc 10500 ccacccacga ggaacatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg 10560 gattgatgtg atggtcaaca tggtggagca cgacactctc gtctactcca agaatatcaa 10620 agatacagtc tcagaagacc agagggctat tgagactttt caacaaaggg taatatcggg 10680 aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga cagtagaaaa 10740 ggaagatggc ttctacaaat gccatcattg cgataaagga aaggctatcg ttcaagatgc 10800 ctctaccgac agtggtccca aagatggacc cccacccacg aggaacatcg tggaaaaaga 10860 agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca ctgacgtaag 10920 ggatgacgca caatcccact atccttcgca agacccttcc tctatataag gaagttcatt 10980 tcatttggag agg 10993 <210> 2 <211> 10643 <212> DNA <213> Artificial Sequence <220> <223> pCkpaR2 <400> 2 gaattctcta gaaggcctcc atggggatcc gtttatttac aagagcgtac ggttcaaccc 60 ctgcctcccc tgtaaaactc cctagactga aaactttctt tctagtatct tttctatggc 120 ttcccttgcc cccacattct cactagccaa tcttttgaac ggtagttacg gtgtcgacac 180 tcccgaggat gtggaacgtt tgcgatctga gcaacgtgaa gaggctgctg cggcctgtcg 240 taactacagg cccctacccg ctgtggatgt cagcgagagt gtcacagagg acgcacattc 300 cctccgaact cctgacggag ctcccgctga agcggtgtct gatgagtttg taacttatgg 360 tgctgaagat taccttgaaa aatctgatga tgagctcctt gtcgcttttg agacgatggt 420 caaacccatg cgtatcggac aactatggtg ccccgcgttt aataaatgtt cttttatttc 480 cagcattgct atggccagag ctttgttgtt ggcacctaga acatccaacc gaaccatgaa 540 gtgttttgaa gacctggtcg cggctattta cactaaatcc gatttctatt atggtgaaga 600 gtgtgaagcc gacgacgttc agatggatat ctcgtctcgc gatgtacccg gttattcttt 660 cgaaccgtgg tcccgaacgt ctggattcga accgccgccc atttgtgaag cgtgcgacat 720 gatcatgtac cagtgcccgt gttttgattt caatgcttta aagaaatcgt gcgctgagag 780 gactttcgct gatgactatg ttattgaagg tttagatggt gttgttgata atgcgactct 840 gttgtcgaat ttgggtccat ttttggtacc cgtgaagtgt caatatgaaa aatgtccaac 900 accaaccctc gcgattcctc cagatttaaa tcgtgctact gatcgtgttg atattaattt 960 agttcaatcc atttgtgact cgactctgcc cactcatagt aattacgacg actcttttca 1020 tcaagtgttt gtcgaaagtg cagactactc tatagatctg gatcatgtta gacttcgaca 1080 gtctgatctt atcgcaaaaa ttccagattc agggcatatg ataccggttt tgaacaccgg 1140 gagcggtcac aagagagtag gtacaacgaa ggaggtcctt acagcaatta agaaacgtaa 1200 tgctgacgtt ccagagctag gtgattccgt taatctgtct agattgagta aagctgtggc 1260 tgagagattc ttcatttcat acatcaatgg taactctcta gcgtccagta actttgtcaa 1320 tgtcgttagt aacttccacg attacatgga aaaatggaag tcctcaggtc tttcttatga 1380 tgatctccca gatcttcatg ctgagaatct gcagttttat gatcacatga taaaatctga 1440 tgtgaagcct gtggtgagcg acacactcaa tatcgacaga ccggttccag ctactataac 1500 gtatcataag aagagtataa cctcccagtt ctcaccgtta ttcacagcgc tatttgagcg 1560 cttccagaga tgccttcgag aacgtattat tcttcctgtt ggtaagattt catcccttga 1620 gatggcagga tttgatgtca aaaacaagca ctgcctcgaa attgatctgt ctaagtttga 1680 taaatctcag ggtgaattcc atttgctaat tcaggaacat attttgaatg gtctaggatg 1740 tccagctccg ataactaagt ggtggtgtga ttttcaccga ttctcttaca ttagagaccg 1800 tagagctggt gttggtatgc ctattagttt ccagagacga actggtgatg cattcactta 1860 ttttggcaat accattgtca ccatggcaga gtttgcctgg tgttacgaca ccgaccaatt 1920 cgaaaagctt ttattctcgg gcgatgactc tctaggattt tcattgcttc cccctgttgg 1980 cgatccgagt aaattcacga ctcttttcaa catggaagct aaggtgatgg aacctgcagt 2040 accatatatt tgttcgaagt tcttactctc tgacgagttc ggtaatacat tttccgttcc 2100 agatccattg cgcgaggttc agcggttagg aacaaagaaa attccctatt ctgacaatga 2160 tgaattcttg tttgctcact tcatgagctt tgttgatcga ttgaagtttt tggaccgaat 2220 gtctcagtcg tgcatcgatc agctttcgat tttctttgaa ttgaaataca agaagtctgg 2280 ggaagaggct gctttaatgt taggcgcctt taagaaatat accgctaatt ttcagtccta 2340 caaggaactc tattatttag atcgtcgtca gtgcgaattg atcaattcgt tttgtagtac 2400 agagttcagg gttgagcgtg taaattccaa taaacagcga aagaaatatg gaattgaacg 2460 taggtgcaat gacaaacgtc gaactccaac tggctcgtat ggtggaggcg aagaagcaga 2520 gacgaagatc tcacaagcag aatcgacggg aacgaggtca caaaagtccc agcgagagag 2580 cgcgttcaaa tctcagactg ttccgcttcc taccgttcta tcaagtagat ggttcggaac 2640 tgatagggtc ggaccgcctt gtgaacgcgg cggagttacc cgagcctgag gcctctcgtt 2700 tagagttatc ggcggaagac catgagtttg acgatacgga ttggttcgcc ggtaacgaat 2760 gggcggaagg tgctttctga aacctcccct tccgcaactc cctccggtca cgtggcggga 2820 gctgagttgg cagtattgct ataaactgtc tgaagtcact aaacacattg tggtgaacgg 2880 gttgtccatc cagcttacgg ctaaaatggt cagtcgtaga gaaatctaca ccagcagact 2940 tgcaagtctc tgaggcacct ttgaaaccat ctcctaggtt tcttcggaag gactccggtc 3000 cgtgtacttc tagcacaacg tgctagtttt agggtacggg tgccccccca ctttcgtggg 3060 gcctctaaag gagaccaact agtgtctgta cttatatcag tacactgacg agtccctaaa 3120 ggacgaaacg ggagaacgct agccaccacc accaccacca cgtgtgaatt acaggtgacc 3180 agctcgaatt tccccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct 3240 gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata 3300 attaacatgt aatgcatgac gttatttatg agatgggttt ttatgattag agtcccgcaa 3360 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg 3420 cgcgcggtgt catctatgtt actagatcgg gaattaaact atcagtgttt gacaggatat 3480 attggcgggt aaacctaaga gaaaagagcg tttattagaa taacggatat ttaaaagggc 3540 gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg gttcccctcg 3600 ggatcaaagt actttgatcc aacccctccg ctgctatagt gcagtcggct tctgacgttc 3660 agtgcagccg tcttctgaaa acgacatgtc gcacaagtcc taagttacgc gacaggctgc 3720 cgccctgccc ttttcctggc gttttcttgt cgcgtgtttt agtcgcataa agtagaatac 3780 ttgcgactag aaccggagac attacgccat gaacaagagc gccgccgctg gcctgctggg 3840 ctatgcccgc gtcagcaccg acgaccagga cttgaccaac caacgggccg aactgcacgc 3900 ggccggctgc accaagctgt tttccgagaa gatcaccggc accaggcgcg accgcccgga 3960 gctggccagg atgcttgacc acctacgccc tggcgacgtt gtgacagtga ccaggctaga 4020 ccgcctggcc cgcagcaccc gcgacctact ggacattgcc gagcgcatcc aggaggccgg 4080 cgcgggcctg cgtagcctgg cagagccgtg ggccgacacc accacgccgg ccggccgcat 4140 ggtgttgacc gtgttcgccg gcattgccga gttcgagcgt tccctaatca tcgaccgcac 4200 ccggagcggg cgcgaggccg ccaaggcccg aggcgtgaag tttggccccc gccctaccct 4260 caccccggca cagatcgcgc acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa 4320 agaggcggct gcactgcttg gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag 4380 cgaggaagtg acgcccaccg aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac 4440 cgaggccgac gccctggcgg ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg 4500 caccaggacg gccaggacga accgtttttc attaccgaag agatcgaggc ggagatgatc 4560 gcggccgggt acgtgttcga gccgcccgcg cacgtctcaa ccgtgcggct gcatgaaatc 4620 ctggccggtt tgtctgatgc caagctggcg gcctggccgg ccagcttggc cgctgaagaa 4680 accgagcgcc gccgtctaaa aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg 4740 gtcgctgcgt atatgatgcg atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg 4800 ttatcgctgt acttaaccag aaaggcgggt caggcaagac gaccatcgca acccatctag 4860 cccgcgccct gcaactcgcc ggggccgatg ttctgttagt cgattccgat ccccagggca 4920 gtgcccgcga ttgggcggcc gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc 4980 gcccgacgat tgaccgcgac gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg 5040 gagcgcccca ggcggcggac ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga 5100 ttccggtgca gccaagccct tacgacatat gggccaccgc cgacctggtg gagctggtta 5160 agcagcgcat tgaggtcacg gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga 5220 tcaaaggcac gcgcatcggc ggtgaggttg ccgaggcgct ggccgggtac gagctgccca 5280 ttcttgagtc ccgtatcacg cagcgcgtga gctacccagg cactgccgcc gccggcacaa 5340 ccgttcttga atcagaaccc gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg 5400 aaattaaatc aaaactcatt tgagttaatg aggtaaagag aaaatgagca aaagcacaaa 5460 cacgctaagt gccggccgtc cgagcgcacg cagcagcaag gctgcaacgt tggccagcct 5520 ggcagacacg ccagccatga agcgggtcaa ctttcagttg ccggcggagg atcacaccaa 5580 gctgaagatg tacgcggtac gccaaggcaa gaccattacc gagctgctat ctgaatacat 5640 cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag tagatgaatt ttagcggcta 5700 aaggaggcgg catggaaaat caagaacaac caggcaccga cgccgtggaa tgccccatgt 5760 gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt tgtctgccgg ccctgcaatg 5820 gcactggaac ccccaagccc gaggaatcgg cgtgacggtc gcaaaccatc cggcccggta 5880 caaatcggcg cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc 5940 cagcggcaac gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat 6000 cgaatccgca aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg 6060 cccaagggcg acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc 6120 gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct 6180 ggcgaggtga tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc 6240 ggcatggcca gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa 6300 tccatgaacc gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac 6360 gttgcggacg tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg 6420 gtagaaacct gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc 6480 aagaacggcc gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc 6540 gtaaagagcg aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac 6600 cgcgagatca cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg 6660 atcgatcccg gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca 6720 gaagccagat ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag 6780 aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg 6840 aaggaggagg cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag 6900 ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca 6960 ggggaaaaag gtcgaaaagg tctctttcct gtggatagca cgtacattgg gaacccaaag 7020 ccgtacattg ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg 7080 tcacacatgt aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct 7140 ttaaaactta ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc 7200 acagccgaag agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc 7260 gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc 7320 aatctaccag ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa 7380 ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc 7440 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc 7500 gtcagcgggt gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg 7560 agtgtatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg 7620 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct 7680 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 7740 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 7800 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 7860 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 7920 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 7980 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 8040 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 8100 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 8160 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 8220 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 8280 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 8340 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 8400 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 8460 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat 8520 tctaggtact aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc 8580 ttgatcccca gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg 8640 atcgaccgga cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga 8700 tcaataaagc cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg 8760 tgggaaaaga caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc 8820 ggatctttaa atggagtgtc ttcttcccag ttttcgcaat ccacatcggc cagatcgtta 8880 ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc 8940 gatatgtcga tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca 9000 gggctttgtt catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc 9060 agattgctcc agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct 9120 tccagccata gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg 9180 ctgtccgtca tttttaaata taggttttca ttttctccca ccagcttata taccttagca 9240 ggagacattc cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc 9300 ggtgatattc tcattttagc catttattat ttccttcctc ttttctacag tatttaaaga 9360 taccccaaga agctaattat aacaagacga actccaattc actgttcctt gcattctaaa 9420 accttaaata ccagaaaaca gctttttcaa agttgttttc aaagttggcg tataacatag 9480 tatcgacgga gccgattttg aaaccgcggt gatcacaggc agcaacgctc tgtcatcgtt 9540 acaatcaaca tgctaccctc cgcgagatca tccgtgtttc aaacccggca gcttagttgc 9600 cgttcttccg aatagcatcg gtaacatgag caaagtctgc cgccttacaa cggctctccc 9660 gctgacgccg tcccggactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg 9720 ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac 9780 gcttagacaa cttaataaca cattgcggac gtttttaatg tactgaatta acgccgaatt 9840 aattcctagg ccaccatgtt gggcccggcg cgccaagctt gcatgcctgc aggtcaacat 9900 ggtggagcac gacactctcg tctactccaa gaatatcaaa gatacagtct cagaagacca 9960 gagggctatt gagacttttc aacaaagggt aatatcggga aacctcctcg gattccattg 10020 cccagctatc tgtcacttca tcgaaaggac agtagaaaag gaagatggct tctacaaatg 10080 ccatcattgc gataaaggaa aggctatcgt tcaagatgcc tctaccgaca gtggtcccaa 10140 agatggaccc ccacccacga ggaacatcgt ggaaaaagaa gacgttccaa ccacgtcttc 10200 aaagcaagtg gattgatgtg atggtcaaca tggtggagca cgacactctc gtctactcca 10260 agaatatcaa agatacagtc tcagaagacc agagggctat tgagactttt caacaaaggg 10320 taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga 10380 cagtagaaaa ggaagatggc ttctacaaat gccatcattg cgataaagga aaggctatcg 10440 ttcaagatgc ctctaccgac agtggtccca aagatggacc cccacccacg aggaacatcg 10500 tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt gatatctcca 10560 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag 10620 gaagttcatt tcatttggag agg 10643 <210> 3 <211> 9849 <212> DNA <213> Artificial Sequence <220> <223> pCkpaR3 <400> 3 gaattctcta gaaggcctcc atggggatcc gtaatcttac cactgtgtgt gtgcgtgtgt 60 gtgtgtcgag tcgtgttgtc cgcacatttg agtcgtgttg tccgcacatt ttattttatc 120 tttgtacagt gtgttagatt tcccgaggca tggctttcca aggtaccagt aggactttaa 180 ctcaacagtc ctcagcggct acgtctgacg atcttcaaaa gatattattt agccctgaag 240 ccattaagaa aatggctact gagtgtgacc taggccggca tcattggatg cgcgctgata 300 atgctatttc agtccggccc ctcgttcccg aagtaaccca cggtcgtatt gcttccttct 360 ttaaatctgg atatgatgtt ggtgaattgt gctcaaaagg atacatgagc gtccctcaag 420 tgttgtgtgc tgttactcga acggtttcca ctgatgctga agggtccttg agaatttact 480 tagctgatct aggtgacaag gagttatctc ctatagatgg acaatgcgtt tcgttacata 540 accatgatct tcccgctttg gtgtctttcc aaccgacgta tgactgtcct atggaaacag 600 ttgggaatcg caagcggtgt tttgctgtcg ttatcgaaag acatggttac attgggtata 660 ccggtaccac agctagcgtg tgtagtaatt ggcaagcaag gttttcttct aagaataaca 720 actacactca tatcgcagct gggaagactc tagtactgcc tttcaacaga ttagctgagc 780 aaacaaaacc gtcagctgtc gctcgcctgt tgaagtcgca attgaacaac attgaatctt 840 cgcaatatct gttaacgaac gcgaagatta atcagaatgc gcgcagtgag tccgaggaat 900 taaatgttga gagccctccc gccgcaatcg ggagttcttc cgcgtcccgc tccgaagcct 960 tcagaccgca ggtggttaac ggtctttagc actttggtgc gtattagtat ataagtattt 1020 gtgagtctgt acataatact atatctatag tgtcctgtgt gagttgatac agtagacatc 1080 tgtgacgcga tgccgtgttg agaagggaac acatctggtt ttagtaagcc tacatcatag 1140 ttttgaggtt caattcctct tactccctgt tgagcccctt actttctcat ggatgcttct 1200 ccgcgagatt gcgttattgt ctactgacta tttagagagt gtgtgtgctg tgttttctct 1260 tttgtgtcgt agaattgagt cgagtcatgg acaaatctga atcaaccagt gctggtcgta 1320 accgtcgacg tcgtccgcgt cgtggttccc gctccgcccc ctcctccgcg gatgccacct 1380 ttagagtctt gtcgcagcag ctttcgcgac ttaataagac gttgtcagct ggtcgtccaa 1440 ctattaacca cccaaccttt gtagggagtg agcgttgtaa acctggatac acgttcacat 1500 ctattaccgt aaagccacca aaaatagacc gcgggtctta ttatggtaaa aggttgctat 1560 tacctgattc agtcacagaa tatgataaga aacttgtttc gcgcattcaa attcaagtta 1620 atcctttgcc gaaatttgat tctaccgtgt gggtgacagt ccgtaaagtt cctgcctcct 1680 cggacttatc cgttgccgcc atctctgcta tgtttgcgga cggagcctca ccggtactgg 1740 tttatcagta tgccgcatct ggagtccaag ctaacaacaa actgttgtat gatctttcgg 1800 cgatgcgcgc tgatataggc gacatgagaa agtacgccgt cctcgtgtac tcaaaagacg 1860 atgcactcga gacggacgag ctggtacttc atgttgacgt cgagcaccaa cgcattccca 1920 cgtctggggt gctcccagta tgattctgtg ttttccagaa ccctccctcc gatttctgtg 1980 gcgggagctg agttgtcagt tctgctataa actgtctgaa gtcactaaac gttttacggt 2040 gaacgggttg tccatccagc ttacggctaa aatggtcagt cgtggagaaa tccacgccag 2100 cagacttaca agtctctgag gcgcctttga aaccatctcc taggtttctt tggaaggact 2160 tcggtccgtg tacttctagc acaatgtgct agtttcaggg tacgggtgcc cccccacttt 2220 cgtgggggcc tctaaaagga gaccaggatc cggtaccgag ctcacgcgtc tcgaggcccg 2280 ggcactagtg tctgtactta tatcagtaca ctgacgagtc cctaaaggac gaaacgggag 2340 aacgctagcc accaccacca ccaccacgtg tgaattacag gtgaccagct cgaatttccc 2400 cgatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc 2460 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg 2520 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata 2580 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc 2640 tatgttacta gatcgggaat taaactatca gtgtttgaca ggatatattg gcgggtaaac 2700 ctaagagaaa agagcgttta ttagaataac ggatatttaa aagggcgtga aaaggtttat 2760 ccgttcgtcc atttgtatgt gcatgccaac cacagggttc ccctcgggat caaagtactt 2820 tgatccaacc cctccgctgc tatagtgcag tcggcttctg acgttcagtg cagccgtctt 2880 ctgaaaacga catgtcgcac aagtcctaag ttacgcgaca ggctgccgcc ctgccctttt 2940 cctggcgttt tcttgtcgcg tgttttagtc gcataaagta gaatacttgc gactagaacc 3000 ggagacatta cgccatgaac aagagcgccg ccgctggcct gctgggctat gcccgcgtca 3060 gcaccgacga ccaggacttg accaaccaac gggccgaact gcacgcggcc ggctgcacca 3120 agctgttttc cgagaagatc accggcacca ggcgcgaccg cccggagctg gccaggatgc 3180 ttgaccacct acgccctggc gacgttgtga cagtgaccag gctagaccgc ctggcccgca 3240 gcacccgcga cctactggac attgccgagc gcatccagga ggccggcgcg ggcctgcgta 3300 gcctggcaga gccgtgggcc gacaccacca cgccggccgg ccgcatggtg ttgaccgtgt 3360 tcgccggcat tgccgagttc gagcgttccc taatcatcga ccgcacccgg agcgggcgcg 3420 aggccgccaa ggcccgaggc gtgaagtttg gcccccgccc taccctcacc ccggcacaga 3480 tcgcgcacgc ccgcgagctg atcgaccagg aaggccgcac cgtgaaagag gcggctgcac 3540 tgcttggcgt gcatcgctcg accctgtacc gcgcacttga gcgcagcgag gaagtgacgc 3600 ccaccgaggc caggcggcgc ggtgccttcc gtgaggacgc attgaccgag gccgacgccc 3660 tggcggccgc cgagaatgaa cgccaagagg aacaagcatg aaaccgcacc aggacggcca 3720 ggacgaaccg tttttcatta ccgaagagat cgaggcggag atgatcgcgg ccgggtacgt 3780 gttcgagccg cccgcgcacg tctcaaccgt gcggctgcat gaaatcctgg ccggtttgtc 3840 tgatgccaag ctggcggcct ggccggccag cttggccgct gaagaaaccg agcgccgccg 3900 tctaaaaagg tgatgtgtat ttgagtaaaa cagcttgcgt catgcggtcg ctgcgtatat 3960 gatgcgatga gtaaataaac aaatacgcaa ggggaacgca tgaaggttat cgctgtactt 4020 aaccagaaag gcgggtcagg caagacgacc atcgcaaccc atctagcccg cgccctgcaa 4080 ctcgccgggg ccgatgttct gttagtcgat tccgatcccc agggcagtgc ccgcgattgg 4140 gcggccgtgc gggaagatca accgctaacc gttgtcggca tcgaccgccc gacgattgac 4200 cgcgacgtga aggccatcgg ccggcgcgac ttcgtagtga tcgacggagc gccccaggcg 4260 gcggacttgg ctgtgtccgc gatcaaggca gccgacttcg tgctgattcc ggtgcagcca 4320 agcccttacg acatatgggc caccgccgac ctggtggagc tggttaagca gcgcattgag 4380 gtcacggatg gaaggctaca agcggccttt gtcgtgtcgc gggcgatcaa aggcacgcgc 4440 atcggcggtg aggttgccga ggcgctggcc gggtacgagc tgcccattct tgagtcccgt 4500 atcacgcagc gcgtgagcta cccaggcact gccgccgccg gcacaaccgt tcttgaatca 4560 gaacccgagg gcgacgctgc ccgcgaggtc caggcgctgg ccgctgaaat taaatcaaaa 4620 ctcatttgag ttaatgaggt aaagagaaaa tgagcaaaag cacaaacacg ctaagtgccg 4680 gccgtccgag cgcacgcagc agcaaggctg caacgttggc cagcctggca gacacgccag 4740 ccatgaagcg ggtcaacttt cagttgccgg cggaggatca caccaagctg aagatgtacg 4800 cggtacgcca aggcaagacc attaccgagc tgctatctga atacatcgcg cagctaccag 4860 agtaaatgag caaatgaata aatgagtaga tgaattttag cggctaaagg aggcggcatg 4920 gaaaatcaag aacaaccagg caccgacgcc gtggaatgcc ccatgtgtgg aggaacgggc 4980 ggttggccag gcgtaagcgg ctgggttgtc tgccggccct gcaatggcac tggaaccccc 5040 aagcccgagg aatcggcgtg acggtcgcaa accatccggc ccggtacaaa tcggcgcggc 5100 gctgggtgat gacctggtgg agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat 5160 cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga 5220 atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg aagccgccca agggcgacga 5280 gcaaccagat tttttcgttc cgatgctcta tgacgtgggc acccgcgata gtcgcagcat 5340 catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg 5400 ctacgagctt ccagacgggc acgtagaggt ttccgcaggg ccggccggca tggccagtgt 5460 gtgggattac gacctggtac tgatggcggt ttcccatcta accgaatcca tgaaccgata 5520 ccgggaaggg aagggagaca agcccggccg cgtgttccgt ccacacgttg cggacgtact 5580 caagttctgc cggcgagccg atggcggaaa gcagaaagac gacctggtag aaacctgcat 5640 tcggttaaac accacgcacg ttgccatgca gcgtacgaag aaggccaaga acggccgcct 5700 ggtgacggta tccgagggtg aagccttgat tagccgctac aagatcgtaa agagcgaaac 5760 cgggcggccg gagtacatcg agatcgagct agctgattgg atgtaccgcg agatcacaga 5820 aggcaagaac ccggacgtgc tgacggttca ccccgattac tttttgatcg atcccggcat 5880 cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc aaggcagaag ccagatggtt 5940 gttcaagacg atctacgaac gcagtggcag cgccggagag ttcaagaagt tctgtttcac 6000 cgtgcgcaag ctgatcgggt caaatgacct gccggagtac gatttgaagg aggaggcggg 6060 gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc 6120 cggttcctaa tgtacggagc agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg 6180 aaaaggtctc tttcctgtgg atagcacgta cattgggaac ccaaagccgt acattgggaa 6240 ccggaacccg tacattggga acccaaagcc gtacattggg aaccggtcac acatgtaagt 6300 gactgatata aaagagaaaa aaggcgattt ttccgcctaa aactctttaa aacttattaa 6360 aactcttaaa acccgcctgg cctgtgcata actgtctggc cagcgcacag ccgaagagct 6420 gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc 6480 tatcgcggcc gctggccgct caaaaatggc tggcctacgg ccaggcaatc taccagggcg 6540 cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca catcaaggca ccctgcctcg 6600 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag 6660 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg 6720 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct 6780 taactatgcg gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc 6840 gcacagatgc gtaaggagaa aataccgcat caggcgctct tccgcttcct cgctcactga 6900 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat 6960 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca 7020 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc 7080 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata 7140 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc 7200 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc 7260 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga 7320 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc 7380 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag 7440 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag 7500 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag 7560 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca 7620 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga 7680 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgcattcta ggtactaaaa 7740 caattcatcc agtaaaatat aatattttat tttctcccaa tcaggcttga tccccagtaa 7800 gtcaaaaaat agctcgacat actgttcttc cccgatatcc tccctgatcg accggacgca 7860 gaaggcaatg tcataccact tgtccgccct gccgcttctc ccaagatcaa taaagccact 7920 tactttgcca tctttcacaa agatgttgct gtctcccagg tcgccgtggg aaaagacaag 7980 ttcctcttcg ggcttttccg tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg 8040 agtgtcttct tcccagtttt cgcaatccac atcggccaga tcgttattca gtaagtaatc 8100 caattcggct aagcggctgt ctaagctatt cgtataggga caatccgata tgtcgatgga 8160 gtgaaagagc ctgatgcact ccgcatacag ctcgataatc ttttcagggc tttgttcatc 8220 ttcatactct tccgagcaaa ggacgccatc ggcctcactc atgagcagat tgctccagcc 8280 atcatgccgt tcaaagtgca ggacctttgg aacaggcagc tttccttcca gccatagcat 8340 catgtccttt tcccgttcca catcataggt ggtcccttta taccggctgt ccgtcatttt 8400 taaatatagg ttttcatttt ctcccaccag cttatatacc ttagcaggag acattccttc 8460 cgtatctttt acgcagcggt atttttcgat cagttttttc aattccggtg atattctcat 8520 tttagccatt tattatttcc ttcctctttt ctacagtatt taaagatacc ccaagaagct 8580 aattataaca agacgaactc caattcactg ttccttgcat tctaaaacct taaataccag 8640 aaaacagctt tttcaaagtt gttttcaaag ttggcgtata acatagtatc gacggagccg 8700 attttgaaac cgcggtgatc acaggcagca acgctctgtc atcgttacaa tcaacatgct 8760 accctccgcg agatcatccg tgtttcaaac ccggcagctt agttgccgtt cttccgaata 8820 gcatcggtaa catgagcaaa gtctgccgcc ttacaacggc tctcccgctg acgccgtccc 8880 ggactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct 8940 gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta 9000 ataacacatt gcggacgttt ttaatgtact gaattaacgc cgaattaatt cctaggccac 9060 catgttgggc ccggcgcgcc aagcttgcat gcctgcaggt caacatggtg gagcacgaca 9120 ctctcgtcta ctccaagaat atcaaagata cagtctcaga agaccagagg gctattgaga 9180 cttttcaaca aagggtaata tcgggaaacc tcctcggatt ccattgccca gctatctgtc 9240 acttcatcga aaggacagta gaaaaggaag atggcttcta caaatgccat cattgcgata 9300 aaggaaaggc tatcgttcaa gatgcctcta ccgacagtgg tcccaaagat ggacccccac 9360 ccacgaggaa catcgtggaa aaagaagacg ttccaaccac gtcttcaaag caagtggatt 9420 gatgtgatgg tcaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat 9480 acagtctcag aagaccagag ggctattgag acttttcaac aaagggtaat atcgggaaac 9540 ctcctcggat tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa 9600 gatggcttct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct 9660 accgacagtg gtcccaaaga tggaccccca cccacgagga acatcgtgga aaaagaagac 9720 gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga cgtaagggat 9780 gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat 9840 ttggagagg 9849 <210> 4 <211> 26 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 4 ggaattctgg tctccttttr gagrcc 26 <210> 5 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 5 agggatcctg gtctcctttt rgagrcc 27 <210> 6 <211> 29 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 6 aggatccgtt tatttacaag agcgtacgg 29 <210> 7 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 7 agggatcctg gtctcctttt rgagrcc 27 <210> 8 <211> 30 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 8 agggatccgt ttatttacaa gagcgtacgg 30 <210> 9 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 9 gactagtggt ctccttttrg agrcc 25 <210> 10 <211> 25 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 10 agggatccgt aatcttacca ctgtg 25 <210> 11 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 11 agggatcctg gtctcctttt rgagrcc 27 <210> 12 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 12 tccctgttga gcccccttac ttt 23 <210> 13 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> primer <400> 13 caactcagat cccgccacag a 21

Claims (3)

Agrobacterium Tumefaciens under accession number KACC95081P. Agrobacterium tumefaciens with accession number KACC95082P. Agrobacterium tumefaciens with accession number KACC95083P.

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