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植物相关质粒

PGR107 植物载体, VIGS vector
发布时间:2018-05-28 16:53:38 | 浏览次数:



测序及更多信息: 链接:https://pan.baidu.com/s/1tmR2uUzJbFO9zwkAIrjygg 
提取码:eqvq 

采购地址:淘宝店铺地址

载体基本信息

载体名称:

pGR107

质粒类型:

植物载体, VIGS vector

启动子:

-

表达水平:

克隆方法:

多克隆位点,限制性内切酶

载体大小:

10436 bp

5' 测序引物:  

-

3' 测序引物:

-

载体标签:

/

载体抗性:

KAN

筛选标记:

-

稳定性:

稳表达

组成型:

组成型 Constitutive

病毒/非病毒:

非病毒



PGR107 polylinker sequence:


 

TGCAAACTAGATGCAGAAACCATAAGGGCCATTGCCGATCTCAAGCCACTCTCCGTTGAACGGTTAAGTTTCCATTGATACTCGAAAGAGGTCAGCACCAGCTAGCATCGATTCCCGGGTCGACCGCCGATGAACGGTTAAGTTTCCATTGATACTCGAAAGATGTCAGCACCAGCTAGCACAACACAGCCCATAGGGTCAACTACCTCAACTACCACAAAAACTGC

Plasmids, strains and plant

Both pgR106 and pgR107 are binary vectors based on pGreen0000, they are basically the same vector except that the cloning site in pgR106 contains ClaI-AscI-NotI-SalI, while in pgR107 contains CalI-SmalI-SalI. AscI and NotI are rare cutting enzymes in that both of them are 8-basepair cutting enzymes and all the 8 base pairs consist of only GC, which will ensure less truncation for cDNA cloning. Therefore pgR106 is more suitable for cDNA library construction, while pgR107 is more suitable for PCR product cloning considering the SmaI site in its cloning site. The selection marker for pgR106 and pgR107 is kanamycin (50ug/ml). Both of them can replicate to high copy number in E.coli. But in agrobaterium they need helper plasmid pJIC Sa_Rep ,which carrys tetracycline as selection mark (5ug/ml), for replicating.

 


We recommend agrobacterium strain GV3101 to work with these two vectors because of its faster growth and high transformation efficiency, though other agrobacterium strains can also work very well with pgR106/pgR107.

Nicotianan benthamiana has been proved to be a suitable plant for easy inoculation and successful VIGS using these two vectors.

Cloning of target gene into VIGS vector

Just like normal cloning, nothing really special in this step. After transformation of E.coli., use kanamycin (50ug/ml) plate for positive colony selection. Then prepare plasmid from confirmed positive colonies and go to next step.

Transformation of agrobacterium

You can either use your constructs and helper plasmid pJIC SA_Rep together to do co-transformation of GV3101 or use your constructs only to transform GV3101 already containing pJIC SA_Rep. The transformation efficiency is much higher in the second method. Since pgR106/107 can not replicate in agrobacterium without pJIC SA_Rep, tetracycline is optional in the selective plate. But it is advisable to use both kanamycin and tetracycline as selection mark to prevent kanamycin resistant strains from contaminating.

The positive agrobacterium colony should be reconfirmed by PCR before inoculation. just to be sure the target gene insert is still there.

Toothpick inoculation of target plant

Once you get agrobacterium colonies containing VIGS constructs on the plate , pick up the colonies and stab target plant leaves on the main vein several times. Choose plants with size of about 5-6 cm in diameter for inoculation. Alternatively , you can infiltrate the agrobacterium culture into plant leaves, which normally gives stronger inoculum.


全序列

1        TTTTTATCCC CGGAAGCCTG TGGATAGAGG GTAGTTATCC ACGTGAAACC GCTAATGCCC
61       CGCAAAGCCT TGATTCACGG GGCTTTCCGG CCCGCTCCAA AAACTATCCA CGTGAAATCG
121      CTAATCAGGG TACGTGAAAT CGCTAATCGG AGTACGTGAA ATCGCTAATA AGGTCACGTG
181      AAATCGCTAA TCAAAAAGGC ACGTGAGAAC GCTAATAGCC CTTTCAGATC AACAGCTTGC
241      AAACACCCCT CGCTCCGGCA AGTAGTTACA GCAAGTAGTA TGTTCAATTA GCTTTTCAAT
301      TATGAATATA TATATCAATT ATTGGTCGCC CTTGGCTTGT GGACAATGCG CTACGCGCAC
361      CGGCTCCGCC CGTGGACAAC CGCAAGCGGT TGCCCACCGT CGAGCGCCAG CGCCTTTGCC
421      CACAACCCGG CGGCCGGCCG CAACAGATCG TTTTATAAAT TTTTTTTTTT GAAAAAGAAA
481      AAGCCCGAAA GGCGGCAACC TCTCGGGCTT CTGGATTTCC GATCCCCGGA ATTAGATCTT
541      GGCAGGATAT ATTGTGGTGT AACGTTCTCG CATGCCTGCA GGTCAACATG GTGGAGCACG
601      ACACGCTTGT CTACTCCAAA AATATCAAAG ATACAGTCTC AGAAGACCAA AGGGCAATTG
661      AGACTTTTCA ACAAAGGGTA ATATCCGGAA ACCTCCTCGG ATTCCATTGC CCAGCTATCT
721      GTCACTTTAT TGTGAAGATA GTGGAAAAGG AAGGTGGCTC CTACAAATGC CATCATTGCG
781      ATAAAGGAAA GGCCATCGTT GAAGATGCCT CTGCCGACAG TGGTCCCAAA GATGGACCCC
841      CACCCACGAG GAGCATCGTG GAAAAAGAAG ACGTTCCAAC CACGTCTTCA AAGCAAGTGG
901      ATTGATGTGA TATCTCCACT GACGTAAGGG ATGACGCACA ATCCCACTAT CCTTCGCAAG
961      ACCCTTCCTC TATATAAGGA AGTTCATTTC ATTTGGAGAG GAGAAAACTA AACCATACAC
1021     CACCAACACA ACCAAACCCA CCACGCCCAA TTGTTACACA CCCGCTTGAA AAAGAAAGTT
1081     TAACAAATGG CCAAGGTGCG CGAGGTTTAC CAATCTTTTA CAGACTCCAC CACAAAAACT
1141     CTCATCCAAG ATGAGGCTTA TAGAAACATT CGCCCCATCA TGGAAAAACA CAAACTAGCT
1201     AACCCTTACG CTCAAACGGT TGAAGCGGCT AATGATCTAG AGGGGTTCGG CATAGCCACC
1261     AATCCCTATA GCATTGAATT GCATACACAT GCAGCCGCTA AGACCATAGA GAATAAACTT
1321     CTAGAGGTGC TTGGTTCCAT CCTACCACAA GAACCTGTTA CATTTATGTT TCTTAAACCC
1381     AGAAAGCTAA ACTACATGAG AAGAAACCCG CGGATCAAGG ACATTTTCCA AAATGTTGCC
1441     ATTGAACCAA GAGACGTAGC CAGGTACCCC AAGGAAACAA TAATTGACAA ACTCACAGAG
1501     ATCACAACGG AAACAGCATA CATTAGTGAC ACTCTGCACT TCTTGGATCC GAGCTACATA
1561     GTGGAGACAT TCCAAAACTG CCCAAAATTG CAAACATTGT ATGCGACCTT AGTTCTCCCC
1621     GTTGAGGCAG CCTTTAAAAT GGAAAGCACT CACCCGAACA TATACAGCCT CAAATACTTC
1681     GGAGATGGTT TCCAGTATAT ACCAGGCAAC CATGGTGGCG GGGCATACCA TCATGAATTC
1741     GCTCATCTAC AATGGCTCAA AGTGGGAAAG ATCAAGTGGA GGGACCCCAA GGATAGCTTT
1801     CTCGGACATC TCAATTACAC GACTGAGCAG GTTGAGATGC ACACAGTGAC AGTACAGTTG
1861     CAGGAATCGT TCGCGGCAAA CCACTTGTAC TGCATCAGGA GAGGAGACTT GCTCACACCG
1921     GAGGTGCGCA CTTTCGGCCA ACCTGACAGG TACGTGATTC CACCACAGAT CTTCCTCCCA
1981     AAAGTTCACA ACTGCAAGAA GCCGATTCTC AAGAAAACTA TGATGCAGCT CTTCTTGTAT
2041     GTTAGGACAG TCAAGGTCGC AAAAAATTGT GACATTTTTG CCAAAGTCAG ACAATTAATT
2101     AAATCATCTG ACTTGGACAA ATACTCTGCT GTGGAACTGG TTTACTTAGT AAGCTACATG
2161     GAGTTCCTTG CCGATTTACA AGCTACCACC TGCTTCTCAG ACACACTTTC TGGTGGCTTG
2221     CTAACAAAGA CCCTTGCACC GGTGAGGGCT TGGATACAAG AGAAAAAGAT GCAGCTGTTT
2281     GGTCTTGAGG ACTACGCGAA GTTAGTCAAA GCAGTTGATT TCCACCCGGT GGATTTTTCT
2341     TTCAAAGTGG AAACTTGGGA CTTCAGATTC CACCCCTTGC AAGCGTGGAA AGCCTTCCGA
2401     CCAAGGGAAG TGTCGGATGT AGAGGAAATG GAAAGTTTGT TCTCAGATGG GGACCTGCTT
2461     GATTGCTTCA CAAGAATGCC AGCTTATGCG GTAAACGCAG AGGAAGATTT AGCTGCAATC
2521     AGGAAAACGC CCGAGATGGA TGTCGGTCAA GAAGTTAAAG AGCCTGCAGG AGACAGAAAT
2581     CAATACTCAA ACCCTGCAGA AACTTTCCTC AACAAGCTCC ACAGGAAACA CAGTAGGGAG
2641     GTGAAACACC AGGCCGCAAA GAAAGCTAAA CGCCTAGCTG AAATCCAGGA GTCAATGAGA
2701     GCTGAAGGTG ATGCCGAACC AAATGAAATA AGCGGGACGA TGGGGGCAAT ACCCAGCAAC
2761     GCCGAACTTC CTGGCACGAA TGATGCCAGA CAAGAACTCA CACTCCCAAC CACTAAACCT
2821     GTCCCTGCAA GGTGGGAAGA TGCTTCATTC ACAGATTCTA GTGTGGAAGA GGAGCAGGTT
2881     AAACTCCTTG GAAAAGAAAC CGTTGAAACA GCGACGCAAC AAGTCATCGA AGGACTTCCT
2941     TGGAAACACT GGATTCCTCA ATTAAATGCT GTTGGATTCA AGGCGCTGGA AATTCAGAGG
3001     GATAGGAGTG GAACAATGAT CATGCCCATC ACAGAAATGG TCTCCGGGCT GGAAAAAGAG
3061     GACTTCCCTG AAGGAACTCC AAAAGAGTTG GCACGAGAAT TGTTCGCTAT GAACAGAAGC
3121     CCTGCCACCA TCCCTTTGGA CCTGCTTAGA GCCAGAGACT ACGGCAGTGA TGTAAAGAAC
3181     AAGAGAATTG GTGCCATCAC AAAGACACAG GCAACGAGTT GGGGCGAATA CTTGACAGGA
3241     AAGATAGAAA GCTTAACTGA GAGGAAAGTT GCGACTTGTG TCATTCATGG AGCTGGAGGT
3301     TCTGGAAAAA GTCATGCCAT CCAGAAGGCA TTGAGAGAAA TTGGCAAGGG CTCGGACATC
3361     ACTGTAGTCC TGCCGACCAA TGAACTGCGG CTAGATTGGA GTAAGAAAGT GCCTAACACT
3421     GAGCCCTATA TGTTCAAGAC CTCTGAAAAG GCGTTAATTG GGGGAACAGG CAGCATAGTC
3481     ATCTTTGACG ATTACTCAAA ACTTCCTCCC GGTTACATAG AAGCCTTAGT CTGTTTCTAC
3541     TCTAAAATCA AGCTAATCAT TCTAACAGGA GATAGCAGAC AAAGCGTCTA CCATGAAACT
3601     GCTGAGGACG CCTCCATCAG GCATTTGGGA CCAGCAACAG AGTACTTCTC AAAATACTGC
3661     CGATACTATC TCAATGCCAC ACACCGCAAC AAGAAAGATC TTGCGAACAT GCTTGGTGTC
3721     TACAGTGAGA GAACGGGAGT CACCGAAATC AGCATGAGCG CCGAGTTCTT AGAAGGAATC
3781     CCAACTTTGG TACCCTCGGA TGAGAAGAGA AAGCTGTACA TGGGCACCGG GAGGAATGAC
3841     ACGTTCACAT ACGCTGGATG CCAGGGGCTA ACTAAGCCGA AGGTACAAAT AGTGTTGGAC
3901     CACAACACCC AAGTGTGTAG CGCGAATGTG ATGTACACGG CACTTTCTAG AGCCACCGAT
3961     AGGATTCACT TCGTGAACAC AAGTGCAAAT TCCTCTGCCT TCTGGGAAAA GTTGGACAGC
4021     ACCCCTTACC TCAAGACTTT CCTATCAGTG GTGAGAGAAC AAGCACTCAG GGAGTACGAG
4081     CCGGCAGAGG CAGAGCCAAT TCAAGAGCCT GAGCCCCAGA CACACATGTG TGTCGAGAAT
4141     GAGGAGTCCG TGCTAGAAGA GTACAAAGAG GAACTCTTGG AAAAGTTTGA CAGAGAGATC
4201     CACTCTGAAT CCCATGGTCA TTCAAACTGT GTCCAAACTG AAGACACAAC CATTCAGTTG
4261     TTTTCGCATC AACAAGCAAA AGATGAGACC CTCCTCTGGG CGACTATAGA TGCGCGGCTC
4321     AAGACCAGCA ATCAAGAAAC AAACTTCCGA GAATTCCTGA GCAAGAAGGA CATTGGGGAC
4381     GTTCTGTTTT TAAACTACCA AAAAGCTATG GGTTTACCCA AAGAGCGTAT TCCTTTTTCC
4441     CAAGAGGTCT GGGAAGCTTG TGCCCACGAA GTACAAAGCA AGTACCTCAG CAAGTCAAAG
4501     TGCAACTTGA TCAATGGGAC TGTGAGACAG AGCCCAGACT TCGATGAAAA TAAGATTATG
4561     GTATTCCTCA AGTCGCAGTG GGTCACAAAG GTGGAAAAAC TAGGTCTACC CAAGATTAAG
4621     CCAGGTCAAA CCATAGCAGC CTTTTACCAG CAGACTGTGA TGCTTTTTGG AACTATGGCT
4681     AGGTACATGC GATGGTTCAG ACAGGCTTTC CAGCCAAAAG AAGTCTTCAT AAACTGTGAG
4741     ACCACGCCAG ATGACATGTC TGCATGGGCC TTGAACAACT GGAATTTCAG CAGACCTAGC
4801     TTGGCTAATG ACTACACAGC TTTCGACCAG TCTCAGGATG GAGCCATGTT GCAATTTGAG
4861     GTGCTCAAAG CCAAACACCA CTGCATACCA GAGGAAATCA TTCAGGCATA CATAGATATT
4921     AAGACTAATG CACAGATTTT CCTAGGCACG TTATCAATTA TGCGCCTGAC TGGTGAAGGT
4981     CCCACTTTTG ATGCAAACAC TGAGTGCAAC ATAGCTTACA CCCATACAAA GTTTGACATC
5041     CCAGCCGGAA CTGCTCAAGT TTATGCAGGA GACGACTCCG CACTGGACTG TGTTCCAGAA
5101     GTGAAGCATA GTTTCCACAG GCTTGAGGAC AAATTACTCC TAAAGTCAAA GCCTGTAATC
5161     ACGCAGCAAA AGAAGGGCAG TTGGCCTGAG TTTTGTGGTT GGCTGATCAC ACCAAAAGGG
5221     GTGATGAAAG ACCCAATTAA GCTCCATGTT AGCTTAAAAT TGGCTGAAGC TAAGGGTGAA
5281     CTCAAGAAAT GTCAAGATTC CTATGAAATT GATCTGAGTT ATGCCTATGA CCACAAGGAC
5341     TCTCTGCATG ACTTGTTCGA TGAGAAACAG TGTCAGGCAC ACACACTCAC TTGCAGAACA
5401     CTAATCAAGT CAGGGAGAGG CACTGTCTCA CTTTCCCGCC TCAGAAACTT TCTTTAACCG
5461     TTAAGTTACC TTAGAGATTT GAATAAGATG GATATTCTCA TCAGTAGTTT GAAAAGTTTA
5521     GGTTATTCTA GGACTTCCAA ATCTTTAGAT TCAGGACCTT TGGTAGTACA TGCAGTAGCC
5581     GGAGCCGGTA AGTCCACAGC CCTAAGGAAG TTGATCCTCA GACACCCAAC ATTCACCGTG
5641     CATACACTCG GTGTCCCTGA CAAGGTGAGT ATCAGAACTA GAGGCATACA GAAGCCAGGA
5701     CCTATTCCTG AGGGCAACTT CGCAATCCTC GATGAGTATA CTTTGGACAA CACCACAAGG
5761     AACTCTAACC AGGCACTTTT TGCTGACCCT TATCAGGCAC CGGAGTTTAG CCTAGAGCCC
5821     CACTTCTACT TGGAAACATC ATTTCGAGTT CCGAGGAAAG TGGCAGATTT GATAGCTGGC
5881     TGTGGCTTCG ATTTCGAGAC CAACTCACCG GAAGAAGGGC ACTTAGAGAT CACTGGCATA
5941     TTCAAAGGGC CCCTACTCGG AAAGGTGATA GCCATTGATG AGGAGTCTGA GACAACACTG
6001     TCCAGGCATG GTGTTGAGTT TGTTAAGCCC TGCCAAGTGA CGGGACTTGA GTTCAAAGTA
6061     GTCACTATTG TGTCTGCCGC ACCAATAGAG GAAATTGGCC AGTCCACAGC TTTCTACAAC
6121     GCTATCACCA GGTCAAAGGG ATTGACATAT GTCCGCGCAG GGCCATAGGC TGACCGCTCC
6181     GGTCAATTCT GAAAAAGTGT ACATAGTATT AGGTCTATCA TTTGCTTTAG TTTCAATTAC
6241     CTTTCTGCTT TCTAGAAATA GCTTACCCCA CGTCGGTGAC AACATTCACA GCTTGCCACA
6301     CGGAGGAGCT TACAGAGACG GCACCAAAGC AATCTTGTAC AACTCCCCAA ATCTAGGGTC
6361     ACGAGTGAGT CTACACAACG GAAAGAACGC AGCATTTGCT GCCGTTTTGC TACTGACTTT
6421     GCTGATCTAT GGAAGTAAAT ACATATCTCA ACGCAATCAT ACTTGTGCTT GTGGTAACAA
6481     TCATAGCAGT CATTAGCACT TCCTTAGTGA GGACTGAACC TTGTGTCATC AAGATTACTG
6541     GGGAATCAAT CACAGTGTTG GCTTGCAAAC TAGATGCAGA AACCATAAGG GCCATTGCCG
6601     ATCTCAAGCC ACTCTCCGTT GAACGGTTAA GTTTCCATTG ATACTCGAAA GAGGTCAGCA
6661     CCAGCTAGCA TCGATTCCCG GGTCGACCGC CGATAAGCTT GATAGGGCCA TTGCCGATCT
6721     CAAGCCACTC TCCGTTGAAC GGTTAAGTTT CCATTGATAC TCGAAAGATG TCAGCACCAG
6781     CTAGCACAAC ACAGCCCATA GGGTCAACTA CCTCAACTAC CACAAAAACT GCAGGCGCAA
6841     CTCCTGCCAC AGCTTCAGGC CTGTTCACCA TCCCGGATGG GGATTTCTTT AGTACAGCCC
6901     GTGCCATAGT AGCCAGCAAT GCTGTCGCAA CAAATGAGGA CCTCAGCAAG ATTGAGGCTA
6961     TTTGGAAGGA CATGAAGGTG CCCACAGACA CTATGGCACA GGCTGCTTGG GACTTAGTCA
7021     GACACTGTGC TGATGTAGGA TCATCCGCTC AAACAGAAAT GATAGATACA GGTCCCTATT
7081     CCAACGGCAT CAGCAGAGCT AGACTGGCAG CAGCAATTAA AGAGGTGTGC ACACTTAGGC
7141     AATTTTGCAT GAAGTATGCT CCAGTGGTAT GGAACTGGAT GTTAACTAAC AACAGTCCAC
7201     CTGCTAACTG GCAAGCACAA GGTTTCAAGC CTGAGCACAA ATTCGCTGCA TTCGACTTCT
7261     TCAATGGAGT CACCAACCCA GCTGCCATCA TGCCCAAAGA GGGGCTCATC CGGCCACCGT
7321     CTGAAGCTGA AATGAATGCT GCCCAAACTG CTGCCTTTGT GAAGATTACA AAGGCCAGGG
7381     CACAATCCAA CGACTTTGCC AGCCTAGATG CAGCTGTCAC TCGAGGTCGT ATCACTGGAA
7441     CAACAACCGC TGAGGCTGTT GTCACTCTAC CACCACCATA ACTACGTCTA CATAACCGAC
7501     GCCTACCCCA GTTTCATAGT ATTTTCTGGT TTGATTGTAT GAATAATATA AATAAAAAAA
7561     AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA AAAAAAAAAA
7621     AAAAAAAAAA AAAAAAAAAC TAGTGGGTAC CGCGAATTTC CCCGATCGTT CAAACATTTG
7681     GCAATAAAGT TTCTTAAGAT TGAATCCTGT TGCCGGTCTT GCGATGATTA TCATATAATT
7741     TCTGTTGAAT TACGTTAAGC ATGTAATAAT TAACATGTAA TGCATGACGT TATTTATGAG
7801     ATGGGTTTTT ATGATTAGAG TCCCGCAATT ATACATTTAA TACGCGATAG AAAACAAAAT
7861     ATAGCGCGCA AACTAGGATA AATTATCGCG CGCGGTGTCA TCTATGTTAC TAGATCGAAT
7921     TCACTGGCCG TCGTTTTACA ACGTCGTGAC TGGGAAAACC CTGGCGTTAC CCAACTTAAT
7981     CGCCTTGCAG CACATCCCCC TTTCGCCAGC TGGCGTAATA GCGAAGAGGC CCGCACCGAT
8041     CGCCCTTCCC AACAGTTGCG CAGCCTGAAT GGCGAATGGC GCCTGATGAG CTCCAGCTTT
8101     TGTTCCCTTT AGTGAGGGTT AATTCCGAGC TTGGCGTAAT CATGGTCATA GCTGTTTCCT
8161     GTGTGAAATT GTTATCCGCT CACAATTCCA CACAACATAC GAGCCGGAAG CATAAAGTGT
8221     AAAGCCTGGG GTGCCTAATG AGTGAGCTAA CTCACATTAA TTGCGTTGCG CTCACTGCCC
8281     GCTTTCCAGT CGGGAAACCT GTCGTGCCAG CTGCATTAAT GAATCGGCCA ACGCGCGGGG
8341     AGAGGCGGTT TGCGTATTGG GCGCTCTTCC GCTTCCTCGC TCACTGACTC GCTGCGCTCG
8401     GTCGTTCGGC TGCGGCGAGC GGTATCAGCT CACTCAAAGG CGGTAATACG GTTATCCACA
8461     GAATCAGGGG ATAACGCAGG AAAGAACATG AAGGCCTTGA CAGGATATAT TGGCGGGTAA
8521     ACTAAGTCGC TGTATGTGTT TGTTTGAGAT CTCATGTGAG CAAAAGGCCA GCAAAAGGCC
8581     AGGAACCGTA AAAAGGCCGC GTTGCTGGCG TTTTTCCATA GGCTCCGCCC CCCTGACGAG
8641     CATCACAAAA ATCGACGCTC AAGTCAGAGG TGGCGAAACC CGACAGGACT ATAAAGATAC
8701     CAGGCGTTTC CCCCTGGAAG CTCCCTCGTG CGCTCTCCTG TTCCGACCCT GCCGCTTACC
8761     GGATACCTGT CCGCCTTTCT CCCTTCGGGA AGCGTGGCGC TTTCTCATAG CTCACGCTGT
8821     AGGTATCTCA GTTCGGTGTA GGTCGTTCGC TCCAAGCTGG GCTGTGTGCA CGAACCCCCC
8881     GTTCAGCCCG ACCGCTGCGC CTTATCCGGT AACTATCGTC TTGAGTCCAA CCCGGTAAGA
8941     CACGACTTAT CGCCACTGGC AGCAGCCACT GGTAACAGGA TTAGCAGAGC GAGGTATGTA
9001     GGCGGTGCTA CAGAGTTCTT GAAGTGGTGG CCTAACTACG GCTACACTAG AAGGACAGTA
9061     TTTGGTATCT GCGCTCTGCT GAAGCCAGTT ACCTTCGGAA AAAGAGTTGG TAGCTCTTGA
9121     TCCGGCAAAC AAACCACCGC TGGTAGCGGT GGTTTTTTTG TTTGCAAGCA GCAGATTACG
9181     CGCAGAAAAA AAGGATCTCA AGAAGATCCT TTGATCTTTT CTACGGGGTC TGACGCTCAG
9241     TGGAACGAAA ACTCACGTTA AGGGATTTTG GTCATGGTTA CAACCAATTA ACCAATTCTG
9301     ATTAGAAAAA CTCATCGAGC ATCAAATGAA ACTGCAATTT ATTCATATCA GGATTATCAA
9361     TACCATATTT TTGAAAAAGC CGTTTCTGTA ATGAAGGAGA AAACTCACCG AGGCAGTTCC
9421     ATAGGATGGC AAGATCCTGG TATCGGTCTG CGATTCCGAC TCGTCCAACA TCAATACAAC
9481     CTATTAATTT CCCCTCGTCA AAAATAAGGT TATCAAGTGA GAAATCACCA TGAGTGACGA
9541     CTGAATCCGG TGAGAATGGC AAAAGTTTAT GCATTTCTTT CCAGACTTGT TCAACAGGCC
9601     AGCCATTACG CTCGTCATCA AAATCACTCG CATCAACCAA ACCGTTATTC ATTCGTGATT
9661     GCGCCTGAGC GAGACGAAAT ACGCGATCGC TGTTAAAAGG ACAATTACAA ACAGGAATCG
9721     AATGCAACCG GCGCAGGAAC ACTGCCAGCG CATCAACAAT ATTTTCACCT GAATCAGGAT
9781     ATTCTTCTAA TACCTGGAAT GCTGTTTTCC CTGGGATCGC AGTGGTGAGT AACCATGCAT
9841     CATCAGGAGT ACGGATAAAA TGCTTGATGG TCGGAAGAGG CATAAATTCC GTCAGCCAGT
9901     TTAGTCTGAC CATCTCATCT GTAACATCAT TGGCAACGCT ACCTTTGCCA TGTTTCAGAA
9961     ACAACTCTGG CGCATCGGGC TTCCCATACA ATCCATAGAT TGTCGCACCT GATTGCCCGA
10021    CATTATCGCG AGCCCATTTA TACCCATATA AATCAGCATC CATGTTGGAA TTTAATCGCG
10081    GCCTGGAGCA AGACGTTTCC CGTTGAATAT GGCTCATAAC ACCCCTTGTA TTACTGTTTA
10141    TGTAAGCAGA CAGTTTTATT GTTCATGATG ATATATTTTT ATCTTGTGCA ATGTAACATC
10201    AGAGATTTTG AGACACAACG TGGCTTTGTT GAATAAATCG AACTTTTGCT GAGTTGAAGG
10261    ATCAGATCAC GCATCTTCCC GACAACGCAG ACCGTTCCGT GGCAAAGCAA AAGTTCAAAA
10321    TCACCAACTG GTCCACCTAC AACAAAGCTC TCATCAACCG TGGCTCCCTC ACTTTCTGGC
10381    TGGATGATGG GGCGATTCAG GCGATCCCCA TCCAACAGCC CGCCGTCGAG CGGGCT




 
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