质粒名称: | pC015 - dLwCas13a-NF |
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出品公司: | 爱迪基因 |
目录编号: | 91905 |
存储实验室: | Feng Zhang |
载体骨架基本信息 |
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载体名称: | pCAGEN |
载体抗性: | 氨苄西林 |
空载体大小: | 5876 bp |
完整质粒大小: | 10622 bp |
载体类型: | 哺乳细胞表达 |
宿生菌: | NEB Stable |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | dLwCas13a-eGFP-ZF-KRAB |
备用名称: | dead LwCas13a、dead LwC2c2、dCas13a |
物种来源: | H. sapiens (human); Leptotrichia wadei |
插入大小: | 4806 bp |
基因突变: | R474A and R1046A |
启动子 | CAG |
标签/融合蛋白: |
EGFP(插入件上的C端子)
ZF CCR5TC(插入件上的C端子)
KRAB-A(插入件上的C端子)
NLS(插入件上的C端子)
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5' 测序引物 | pCAG-F |
文献引用方法 |
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材料和方法部分: | pC015 - dLwCas13a-NF was a gift from Feng Zhang (Addgene plasmid # 91905 ; http://n2t.net/addgene:91905 ; RRID:Addgene_91905) |
参考文献部分: | RNA targeting with CRISPR-Cas13. Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, Voytas DF, Ting AY, Zhang F. Nature. 2017 Oct 4. doi: 10.1038/nature24049. 10.1038/nature24049 PubMed 28976959 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pC015 - dLwCas13a-NF 10715 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses negative-feedback dLwCas13a for mammalian RNA targeting/imaging. ACCESSION . VERSION . KEYWORDS pC015 - dLwCas13a-NF SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10715) AUTHORS Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, Voytas DF, Ting AY, Zhang F TITLE RNA targeting with CRISPR-Cas13. JOURNAL Nature. 2017 Oct 4. doi: 10.1038/nature24049. PUBMED 28976959 REFERENCE 2 (bases 1 to 10715) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10715 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(281..1141) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 904..923 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1142..1246) /gene="bla" /label=AmpR promoter enhancer 1312..1691 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1694..1969 /label=chicken beta-actin promoter intron 1970..2981 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 2905..2927 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 2989..3008 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 6561..7271 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELY" primer_bind complement(6603..6624) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(6864..6883) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 7211..7232 /label=EGFP-C /note="EGFP, forward primer" CDS 7842..7862 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" intron 7879..8011 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 7994..8013 /label=pMT2-F /note="Synthetic intron, forward primer" misc_feature 8020..8608 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(8073..8093) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(8491..8502) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(8635..8652) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 8641..8752 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(8848..8868) /label=T3 /note="T3 promoter, forward primer" promoter complement(8848..8866) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(8887..8903) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9009..9028) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 9074..9129 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(9128..9147) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(9490..9506) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 9514..9530 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(9538..9568) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9583..9604 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 9663..9858 /label=SV40 promoter /note="SV40 early promoter" rep_origin 9709..9844 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9771..9790 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 9864..9998 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9901..9920) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9955..9974 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(10066..10083) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(10237..110) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10317..10336) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 61 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 121 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 181 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 241 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 301 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 361 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 421 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 481 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 541 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 601 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 661 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 721 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 781 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 841 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 901 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 961 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 1021 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 1081 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 1141 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 1201 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 1261 aagtgccacc tgggtcgaca ttgattattg actagagtca tcctcatcgt cgacattgat 1321 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg 1381 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc 1441 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt 1501 gacgtcaatg ggtggactat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc 1561 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg 1621 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg 1681 ctattaccat gggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc 1741 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg 1801 gggggggggg gggcgcgcgc caggcggggc ggggcggggg cgagggcggg gcggggcgag 1861 gcggagaggt gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc 1921 gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc 1981 gttgccttcg ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact 2041 gaccgcgtta ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta 2101 gcgcttggtt taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct 2161 ccgggagggc cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg 2221 tggggagcgc cgcgtgcggc ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc 2281 ggggctttgt gcgctccgcg tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg 2341 tgcggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag 2401 cagggggtgt gggcgcggcg gtcgggctgt aacccccccc tgcacccccc tccccgagtt 2461 gctgagcacg gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc ggggctcgcc 2521 gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc 2581 ggggagggct cgggggaggg gcgcggcggc cccggagcgc cggcggctgt cgaggcgcgg 2641 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt 2701 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg 2761 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc 2821 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc 2881 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag 2941 cctctgctaa ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg 3001 ttattgtgct gtctcatcat tttggcaaag aattcggtac cgcgggcccg ggcgccacca 3061 tgaaagtgac caaggtcgac ggcatcagcc acaagaagta catcgaagag ggcaagctcg 3121 tgaagtccac cagcgaggaa aaccggacca gcgagagact gagcgagctg ctgagcatcc 3181 ggctggacat ctacatcaag aaccccgaca acgcctccga ggaagagaac cggatcagaa 3241 gagagaacct gaagaagttc tttagcaaca aggtgctgca cctgaaggac agcgtgctgt 3301 atctgaagaa ccggaaagaa aagaacgccg tgcaggacaa gaactatagc gaagaggaca 3361 tcagcgagta cgacctgaaa aacaagaaca gcttctccgt gctgaagaag atcctgctga 3421 acgaggacgt gaactctgag gaactggaaa tctttcggaa ggacgtggaa gccaagctga 3481 acaagatcaa cagcctgaag tacagcttcg aagagaacaa ggccaactac cagaagatca 3541 acgagaacaa cgtggaaaaa gtgggcggca agagcaagcg gaacatcatc tacgactact 3601 acagagagag cgccaagcgc aacgactaca tcaacaacgt gcaggaagcc ttcgacaagc 3661 tgtataagaa agaggatatc gagaaactgt ttttcctgat cgagaacagc aagaagcacg 3721 agaagtacaa gatccgcgag tactatcaca agatcatcgg ccggaagaac gacaaagaga 3781 acttcgccaa gattatctac gaagagatcc agaacgtgaa caacatcaaa gagctgattg 3841 agaagatccc cgacatgtct gagctgaaga aaagccaggt gttctacaag tactacctgg 3901 acaaagagga actgaacgac aagaatatta agtacgcctt ctgccacttc gtggaaatcg 3961 agatgtccca gctgctgaaa aactacgtgt acaagcggct gagcaacatc agcaacgata 4021 agatcaagcg gatcttcgag taccagaatc tgaaaaagct gatcgaaaac aaactgctga 4081 acaagctgga cacctacgtg cggaactgcg gcaagtacaa ctactatctg caagtgggcg 4141 agatcgccac ctccgacttt atcgcccgga accggcagaa cgaggccttc ctgagaaaca 4201 tcatcggcgt gtccagcgtg gcctacttca gcctgaggaa catcctggaa accgagaacg 4261 agaacggtat caccggccgg atgcggggca agaccgtgaa gaacaacaag ggcgaagaga 4321 aatacgtgtc cggcgaggtg gacaagatct acaatgagaa caagcagaac gaagtgaaag 4381 aaaatctgaa gatgttctac agctacgact tcaacatgga caacaagaac gagatcgagg 4441 acttcttcgc caatatcgat gaggccatct ctagtattgc tcacggcatc gtgcatttca 4501 acctggaact ggaaggcaag gacatcttcg ccttcaagaa tatcgccccc agcgagatct 4561 ccaagaagat gtttcagaac gaaatcaacg aaaagaagct gaagctgaaa atcttcaagc 4621 agctgaacag cgccaacgtg ttcaactact acgagaagga tgtgatcatc aagtacctga 4681 agaataccaa gttcaacttc gtgaacaaaa acatcccctt cgtgcccagc ttcaccaagc 4741 tgtacaacaa gattgaggac ctgcggaata ccctgaagtt tttttggagc gtgcccaagg 4801 acaaagaaga gaaggacgcc cagatctacc tgctgaagaa tatctactac ggcgagttcc 4861 tgaacaagtt cgtgaaaaac tccaaggtgt tctttaagat caccaatgaa gtgatcaaga 4921 ttaacaagca gcggaaccag aaaaccggcc actacaagta tcagaagttc gagaacatcg 4981 agaaaaccgt gcccgtggaa tacctggcca tcatccagag cagagagatg atcaacaacc 5041 aggacaaaga ggaaaagaat acctacatcg actttattca gcagattttc ctgaagggct 5101 tcatcgacta cctgaacaag aacaatctga agtatatcga gagcaacaac aacaatgaca 5161 acaacgacat cttctccaag atcaagatca aaaaggataa caaagagaag tacgacaaga 5221 tcctgaagaa ctatgagaag cacaatcgga acaaagaaat ccctcacgag atcaatgagt 5281 tcgtgcgcga gatcaagctg gggaagattc tgaagtacac cgagaatctg aacatgtttt 5341 acctgatcct gaagctgctg aaccacaaag agctgaccaa cctgaagggc agcctggaaa 5401 agtaccagtc cgccaacaaa gaagaaacct tcagcgacga gttggaactg atcaacctgc 5461 tgaacctgga caacaacaga gtgaccgagg acttcgagct ggaagccaac gagatcggca 5521 agttcctgga cttcaacgaa aacaaaatca aggaccggaa agagctgaaa aagttcgaca 5581 ccaacaagat ctatttcgac ggcgagaaca tcatcaagca ccgggccttc tacaatatca 5641 agaaatacgg catgctgaat ctgctggaaa agatcgccga taaggccaag tataagatca 5701 gcctgaaaga actgaaagag tacagcaaca agaagaatga gattgaaaag aactacacca 5761 tgcagcagaa cctgcaccgg aagtacgcca gacccaagaa ggacgaaaag ttcaacgacg 5821 aggactacaa agagtatgag aaggccatcg gcaacatcca gaagtacacc cacctgaaga 5881 acaaggtgga attcaatgag ctgaacctgc tgcagggcct gctgctgaag atcctgcacc 5941 ggctcgtggg ctacaccagc atctgggagc gggacctgag attccggctg aagggcgagt 6001 ttcccgagaa ccactacatc gaggaaattt tcaatttcga caactccaag aatgtgaagt 6061 acaaaagcgg ccagatcgtg gaaaagtata tcaacttcta caaagaactg tacaaggaca 6121 atgtggaaaa gcggagcatc tactccgaca agaaagtgaa gaaactgaag caggagaaga 6181 aagacctgta tattgcaaac tacatcgctc actttaatta tattcctcat gccgagatta 6241 gcctgctgga agtgctggaa aacctgcgga agctgctgtc ctacgaccgg aagctgaaga 6301 acgccatcat gaagtccatc gtggacattc tgaaagaata cggcttcgtg gccaccttca 6361 agatcggcgc tgacaagaag atcgaaatcc agaccctgga atcagagaag atcgtgcacc 6421 tgaagaatct gaagaaaaag aaactgatga ccgaccggaa cagcgaggaa ctgtgcgaac 6481 tcgtgaaagt catgttcgag tacaaggccc tggaaggagg aggtggaagc ggaggaggag 6541 gaagcggagg aggaggtagc gtgagcaagg gcgaggagct gttcaccggg gtggtgccca 6601 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg 6661 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 6721 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct 6781 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 6841 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 6901 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 6961 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg 7021 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg 7081 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 7141 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga 7201 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg 7261 acgagctgta ctctagggga gctggcgctg gagcgggtgc aggggctggc tctagattcc 7321 agtgccggat ctgcatgcgg aacttcagcg accggtccaa cctgagcagg cacatcagaa 7381 cccacaccgg agaaaagccc ttcgcctgcg acatttgcgg ccggaagttc gccatcagca 7441 gcaacctgaa cagccacacc aagatccaca ctggcagcca gaaacctttc cagtgcagaa 7501 tttgtatgag aaactttagc agaagcgaca acctggccag acacatccgg acacatactg 7561 gtgaaaaacc ttttgcctgt gatatctgtg gcagaaagtt tgccacctcc ggcaatctga 7621 cccggcacac aaagattcac ctgcggggca gccagctatc gattgtcgac gctcctgaac 7681 aacgtgaagg tgcttctcaa gtttctgttt ctgttacttt tgaagatgtt gctgttcttt 7741 ttactcgtga tgaatggaaa aaacttgatc tttctcaacg ttctctttat cgtgaagtta 7801 tgcttgaaaa ttattctaat cttgcttcta tggctggatc ccccaagaaa aagcgcaagg 7861 tataactcga ggccgcaggt aagtatcaag gttacaagac aggtttaagg agaccaatag 7921 aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc tattggtctt 7981 actgacatcc actttgcctt tctctccaca ggtgtcgaca atcaacctct ggattacaaa 8041 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac 8101 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc 8161 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt 8221 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc 8281 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc 8341 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg 8401 gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt 8461 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc 8521 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt 8581 cggatctccc tttgggccgc ctccccgcct ggaattcgag ctcggtacga tcagcctcga 8641 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc 8701 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc 8761 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt 8821 gggaagacaa tagcccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta 8881 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctaattcact cctcaggtgc 8941 aggctgccta tcagaaggtg gtggctggtg tggccaatgc cctggctcac aaataccact 9001 gagatcgatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat 9061 ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg 9121 tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat 9181 ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta 9241 taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa 9301 gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca 9361 tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc 9421 ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc 9481 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 9541 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac 9601 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat 9661 ccgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta 9721 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca 9781 gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga 9841 ggcctaggct tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa 9901 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt 9961 tgtccaaact catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca 10021 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc 10081 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg 10141 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa 10201 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga 10261 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag 10321 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct 10381 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg 10441 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc 10501 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt 10561 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta 10621 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac 10681 agtatttggt atctgcgctc tgctgaagcc agtta //