质粒名称: | xCas9(3.7)-ABE(7.10) |
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出品公司: | 爱迪基因 |
目录编号: | 108382 |
存储实验室: | David Liu |
载体骨架基本信息 |
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载体名称: | pCMV |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | -- |
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: | xCas9(3.7)-ABE(7.10),TadA-linker(32aa)-TadA*(7.10)-linker(32aa)-nxCas9(3.7)-NLS |
物种来源: | Synthetic |
插入大小: | 5361 bp |
基因突变: | A262T, R324L, S409I, E480K, E543D, M694I, E1219V |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | NLS(插入件上的N端子) |
克隆方法: | Gibson 克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | CMV Forward |
3' 测序引物 | M13 |
文献引用方法 |
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材料和方法部分: | xCas9(3.7)-ABE(7.10) was a gift from David Liu (Addgene plasmid # 108382 ; http://n2t.net/addgene:108382 ; RRID:Addgene_108382) |
参考文献部分: | Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z, Liu DR. Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155. 10.1038/nature26155 PubMed 29512652 |
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载体基本信息
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载体简介
载体序列
LOCUS xCas9(3.7)-ABE(7.10) 8760 bp ds-DNA circular SYN 09-NOV-2018 DEFINITION Mammalian expression vector for xCas9(3.7)-ABE. ACCESSION . VERSION . KEYWORDS xCas9(3.7)-ABE(7.10) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8760) AUTHORS Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z, Liu DR TITLE Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. JOURNAL Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155. PUBMED 29512652 REFERENCE 2 (bases 1 to 8760) AUTHORS . TITLE Direct Submission JOURNAL Exported Nov 9, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8760 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 19..398 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 399..602 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 553..573 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 644..663 /label=T7 /note="T7 promoter, forward primer" promoter 644..662 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1867..5967 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 6013..6033 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6042..6059 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(6082..6099) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 6088..6312 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6383..6399) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6383..6399) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6396..6418) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6407..6423 /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(6431..6461) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6476..6497 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6614..6631) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6785..7373) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6865..6884) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7544..8404) /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 8167..8186 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8405..8509) /gene="bla" /label=AmpR promoter primer_bind complement(8588..8607) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg 61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc 121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca 181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg 241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg 301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt 361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca 421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg 481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact 541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag 601 ctggtttagt gaaccgtcag atccgctaga gatccgcggc cgctaatacg actcactata 661 gggagagccg ccaccatgtc cgaagtcgag ttttcccatg agtactggat gagacacgca 721 ttgactctcg caaagagggc ttgggatgaa cgcgaggtgc ccgtgggggc agtactcgtg 781 cataacaatc gcgtaatcgg cgaaggttgg aataggccga tcggacgcca cgaccccact 841 gcacatgcgg aaatcatggc ccttcgacag ggagggcttg tgatgcagaa ttatcgactt 901 atcgatgcga cgctgtacgt cacgcttgaa ccttgcgtaa tgtgcgcggg agctatgatt 961 cactcccgca ttggacgagt tgtattcggt gcccgcgacg ccaagacggg tgccgcaggt 1021 tcactgatgg acgtgctgca tcacccaggc atgaaccacc gggtagaaat cacagaaggc 1081 atattggcgg acgaatgtgc ggcgctgttg tccgactttt ttcgcatgcg gaggcaggag 1141 atcaaggccc agaaaaaagc acaatcctct actgactctg gtggttcttc tggtggttct 1201 agcggcagcg agactcccgg gacctcagag tccgccacac ccgaaagttc tggtggttct 1261 tctggtggtt cttccgaagt cgagttttcc catgagtact ggatgagaca cgcattgact 1321 ctcgcaaaga gggctcgaga tgaacgcgag gtgcccgtgg gggcagtact cgtgctcaac 1381 aatcgcgtaa tcggcgaagg ttggaatagg gcaatcggac tccacgaccc cactgcacat 1441 gcggaaatca tggcccttcg acagggaggg cttgtgatgc agaattatcg acttatcgat 1501 gcgacgctgt acgtcacgtt tgaaccttgc gtaatgtgcg cgggagctat gattcactcc 1561 cgcattggac gagttgtatt cggtgttcgc aacgccaaga cgggtgccgc aggttcactg 1621 atggacgtgc tgcattaccc aggcatgaac caccgggtag aaatcacaga aggcatattg 1681 gcggacgaat gtgcggcgct gttgtgttac ttttttcgca tgcccaggca ggtctttaac 1741 gcccagaaaa aagcacaatc ctctactgac tctggtggtt cttctggtgg ttctagcggc 1801 agcgagactc ccgggacctc agagtccgcc acacccgaaa gttctggtgg ttcttctggt 1861 ggttctgaca agaagtactc cattgggctc gctatcggca caaacagcgt cggctgggcc 1921 gtcattacgg acgagtacaa ggtgccgagc aaaaaattca aagttctggg caataccgat 1981 cgccacagca taaagaagaa cctcattggc gccctcctgt tcgactccgg ggagacggcc 2041 gaagccacgc ggctcaaaag aacagcacgg cgcagatata cccgcagaaa gaatcggatc 2101 tgctacctgc aggagatctt tagtaatgag atggctaagg tggatgactc tttcttccat 2161 aggctggagg agtccttttt ggtggaggag gataaaaagc acgagcgcca cccaatcttt 2221 ggcaatatcg tggacgaggt ggcgtaccat gaaaagtacc caaccatata tcatctgagg 2281 aagaagcttg tagacagtac tgataaggct gacttgcggt tgatctatct cgcgctggcg 2341 catatgatca aatttcgggg acacttcctc atcgaggggg acctgaaccc agacaacagc 2401 gatgtcgaca aactctttat ccaactggtt cagacttaca atcagctttt cgaagagaac 2461 ccgatcaacg catccggagt tgacgccaaa gcaatcctga gcgctaggct gtccaaatcc 2521 cggcggctcg aaaacctcat cgcacagctc cctggggaga agaagaacgg cctgtttggt 2581 aatcttatcg ccctgtccct cgggctgacc cccaacttta aatctaactt cgacctggcc 2641 gaagatacca agcttcaact gagcaaagac acctacgatg atgatctcga caatctgctg 2701 gcccagatcg gcgaccagta cgcagacctt tttttggcgg caaagaacct gtcagacgcc 2761 attctgctga gtgatattct gcgagtgaac acggagatca ccaaagctcc gctgagcgct 2821 agtatgatca agctctatga tgagcaccac caagacttga ctttgctgaa ggcccttgtc 2881 agacagcaac tgcctgagaa gtacaaggaa attttcttcg atcagtctaa aaatggctac 2941 gccggataca ttgacggcgg agcaagccag gaggaatttt acaaatttat taagcccatc 3001 ttggaaaaaa tggacggcac cgaggagctg ctggtaaagc ttaacagaga agatctgttg 3061 cgcaaacagc gcactttcga caatggaatc atcccccacc agattcacct gggcgaactg 3121 cacgctatcc tcaggcggca agaggatttc tacccctttt tgaaagataa cagggaaaag 3181 attgagaaaa tcctcacatt tcggataccc tactatgtag gccccctcgc ccggggaaat 3241 tccagattcg cgtggatgac tcgcaaatca gaagagacca tcactccctg gaacttcgag 3301 aaagtcgtgg ataagggggc ctctgcccag tccttcatcg aaaggatgac taactttgat 3361 aaaaatctgc ctaacgaaaa ggtgcttcct aaacactctc tgctgtacga gtacttcaca 3421 gtttataacg agctcaccaa ggtcaaatac gtcacagaag ggatgagaaa gccagcattc 3481 ctgtctggag atcagaagaa agctattgtg gacctcctct tcaagacgaa ccggaaagtt 3541 accgtgaaac agctcaaaga agactatttc aaaaagattg aatgtttcga ctctgttgaa 3601 atcagcggag tggaggatcg cttcaacgca tccctgggaa cgtatcacga tctcctgaaa 3661 atcattaaag acaaggactt cctggacaat gaggagaacg aggacattct tgaggacatt 3721 gtcctcaccc ttacgttgtt tgaagatagg gagatgattg aagaacgctt gaaaacttac 3781 gctcatctct tcgacgacaa agtcatgaag cagctcaaga ggcgccgata tacaggatgg 3841 gggcggctgt caagaaaact gatcaatggg atccgagaca agcagagtgg aaagacaatc 3901 ctggattttc ttaagtccga tggatttgcc aaccggaact tcattcagtt gatccatgat 3961 gactctctca cctttaagga ggacatccag aaagcacaag tttctggcca gggggacagt 4021 cttcacgagc acatcgctaa tcttgcaggt agcccagcta tcaaaaaggg aatactgcag 4081 accgttaagg tcgtggatga actcgtcaaa gtaatgggaa ggcataagcc cgagaatatc 4141 gttatcgaga tggcccgaga gaaccaaacc acccagaagg gacagaagaa cagtagggaa 4201 aggatgaaga ggattgaaga gggtataaaa gaactggggt cccaaatcct taaggaacac 4261 ccagttgaaa acacccagct tcagaatgag aagctctacc tgtactacct gcagaacggc 4321 agggacatgt acgtggatca ggaactggac atcaatcggc tctccgacta cgacgtggat 4381 catatcgtgc cccagtcttt tctcaaagat gattctattg ataataaagt gttgacaaga 4441 tccgataaaa acagagggaa gagtgataac gtcccctcag aagaagttgt caagaaaatg 4501 aaaaattatt ggcggcagct gctgaacgcc aaactgatca cacaacggaa gttcgataat 4561 ctgactaagg ctgaacgagg tggcctgtct gagttggata aagccggttt catcaaaagg 4621 cagcttgttg agacacgcca gatcaccaag cacgtggccc aaattctcga ttcacgcatg 4681 aacaccaagt acgatgaaaa tgacaaactg attcgagagg tgaaagttat tactctgaag 4741 tctaagctgg tctcagattt cagaaaggac tttcagtttt ataaggtgag agagatcaac 4801 aattaccacc atgcgcatga tgcctacctg aatgcagtgg taggcactgc acttatcaaa 4861 aaatatccca agcttgaatc tgaatttgtt tacggagact ataaagtgta cgatgttagg 4921 aaaatgatcg caaagtctga gcaggaaata ggcaaggcca ccgctaagta cttcttttac 4981 agcaatatta tgaatttttt caagaccgag attacactgg ccaatggaga gattcggaag 5041 cgaccactta tcgaaacaaa cggagaaaca ggagaaatcg tgtgggacaa gggtagggat 5101 ttcgcgacag tccggaaggt cctgtccatg ccgcaggtga acatcgttaa aaagaccgaa 5161 gtacagaccg gaggcttctc caaggaaagt atcctcccga aaaggaacag cgacaagctg 5221 atcgcacgca aaaaagattg ggaccccaag aaatacggcg gattcgattc tcctacagtc 5281 gcttacagtg tactggttgt ggctaaagtg gagaaaggga agtctaaaaa actcaaaagc 5341 gtcaaggaac tgctgggcat cacaatcatg gagcgatcaa gcttcgaaaa aaaccccatc 5401 gactttctcg aggcgaaagg atataaagag gtcaaaaaag acctcatcat taagcttccc 5461 aagtactctc tctttgagct tgaaaacggc cggaaacgaa tgctcgctag tgcgggcgtg 5521 ctgcagaaag gtaacgagct ggcactgccc tctaaatacg ttaatttctt gtatctggcc 5581 agccactatg aaaagctcaa agggtctccc gaagataatg agcagaagca gctgttcgtg 5641 gaacaacaca aacactacct tgatgagatc atcgagcaaa taagcgaatt ctccaaaaga 5701 gtgatcctcg ccgacgctaa cctcgataag gtgctttctg cttacaataa gcacagggat 5761 aagcccatca gggagcaggc agaaaacatt atccacttgt ttactctgac caacttgggc 5821 gcgcctgcag ccttcaagta cttcgacact accatagaca gaaagcggta cacctctaca 5881 aaggaggtcc tggacgccac actgattcat cagtcaatta cggggctcta tgaaacaaga 5941 atcgacctct ctcagctcgg tggagacgag ggagctgata agcgcaccgc cgatggttcc 6001 gagttcgaaa gccccaagaa gaagaggaaa gtctaaccgg tcatcatcac catcaccatt 6061 gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt 6121 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 6181 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 6241 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg 6301 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgataccg tcgacctcta 6361 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca 6421 caattccaca caacatacga gccggaagca taaagtgtaa agcctagggt gcctaatgag 6481 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt 6541 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc 6601 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 6661 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 6721 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 6781 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 6841 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 6901 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 6961 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 7021 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 7081 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 7141 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 7201 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag 7261 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 7321 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 7381 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 7441 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt 7501 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca 7561 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg 7621 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac 7681 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg 7741 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc 7801 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta 7861 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac 7921 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc 7981 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac 8041 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact 8101 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa 8161 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt 8221 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca 8281 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa 8341 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac 8401 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg 8461 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc 8521 gaaaagtgcc acctgacgtc gacggatcgg gagatcgatc tcccgatccc ctagggtcga 8581 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg 8641 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga 8701 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac //