质粒名称: | ABE8.8-m |
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出品公司: | addgene |
目录编号: | 136294 |
存储实验室: | Nicole Gaudelli |
载体骨架基本信息 |
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载体名称: | pBR322 |
载体出品公司: | 无 |
空载体大小: | 3400 bp |
完整质粒大小: | 8166 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | ABE8.8-m |
插入大小: | 5300 bp |
基因突变: | D10A in S. pyogenes Cas9, TadA mutations described in manuscript |
标签/融合蛋白: | C-terminal BPNLS (C terminal on insert) |
克隆方法: | Ligation Independent Cloning |
5’测序引物 | T7 |
3’测序引物 | M13R |
文献引用方法 |
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材料和方法部分: | ABE8.8-m was a gift from Nicole Gaudelli (Addgene plasmid # 136294 ; http://n2t.net/addgene:136294 ; RRID:Addgene_136294) |
参考文献部分: |
Directed evolution of adenine base editors with increased activity and therapeutic application. Gaudelli NM, Lam DK, Rees HA, Sola-Esteves NM, Barrera LA, Born DA, Edwards A, Gehrke JM, Lee SJ, Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G. Nat Biotechnol. 2020 Apr 13. pii: 10.1038/s41587-020-0491-6. doi: 10.1038/s41587-020-0491-6. 10.1038/s41587-020-0491-6 PubMed 32284586
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载体基本信息
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载体序列
LOCUS ABE8.8-m 8166 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses ABE8.8-m in mammalian cells. ACCESSION . VERSION . KEYWORDS ABE8.8-m SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8166) AUTHORS Gaudelli NM, Lam DK, Rees HA, Sola-Esteves NM, Barrera LA, Born DA, Edwards A, Gehrke JM, Lee SJ, Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G TITLE Directed evolution of adenine base editors with increased activity and therapeutic application. JOURNAL Nat Biotechnol. 2020 Apr 13. pii: 10.1038/s41587-020-0491-6. doi: 10.1038/s41587-020-0491-6. PUBMED 32284586 REFERENCE 2 (bases 1 to 8166) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..8166 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 111..490 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 491..694 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 645..665 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 736..755 /label=T7 /note="T7 promoter, forward primer" promoter 736..754 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 771..1268 /codon_start=1 /product="DNA adenosine deaminase evolved from the E. coli tRNA adenosine deaminase TadA (Gaudelli et al., 2017)" /label=TadA*(7.10) /translation="SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFG VRNAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLCRFFRMPRRVFNAQKKAQS STD" CDS 1365..5465 /codon_start=1 /product="nickase mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9(D10A) /note="generates RNA-guided single strand nicks in DNA due to the D10A mutation in the RuvC catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 5511..5531 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5540..5557 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(5580..5597) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5586..5810 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(5881..5897) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5881..5897) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5894..5916) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5905..5921 /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(5929..5959) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5974..5995 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6112..6129) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6283..6871) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6363..6382) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7042..7902) /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 7665..7684 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7903..8007) /gene="bla" /label=AmpR promoter primer_bind complement(8086..8105) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga atctgcttag 61 ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt gacattgatt 121 attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga 181 gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg 241 cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg 301 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca 361 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc 421 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc 481 tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc 541 acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa 601 tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag 661 gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta 721 gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg tccgaagtcg 781 agttttccca tgagtactgg atgagacacg cattgactct cgcaaagagg gctcgagatg 841 aacgcgaggt gcccgtgggg gcagtactcg tgctcaacaa tcgcgtaatc ggcgaaggtt 901 ggaatagggc aatcggactc cacgacccca ctgcacatgc ggaaatcatg gcccttcgac 961 agggagggct tgtgatgcag aattatcgac ttatcgatgc gacgctgtac gtcacgtttg 1021 aaccttgcgt aatgtgcgcg ggagctatga ttcactcccg cattggacga gttgtattcg 1081 gtgttcgcaa cgccaagacg ggtgccgcag gttcactgat ggacgtgctg catcatccag 1141 gcatgaacca ccgggtagaa atcacagaag gcatattggc ggacgaatgt gcggcgctgt 1201 tgtgtcgttt ttttcgcatg cccaggcggg tctttaacgc ccagaaaaaa gcacaatcct 1261 ctactgactc tggtggttct tctggtggtt ctagcggcag cgagactccc gggacctcag 1321 agtccgccac acccgaaagt tctggtggtt cttctggtgg ttctgacaag aagtacagca 1381 tcggcctggc catcggcacc aactctgtgg gctgggccgt gatcaccgac gagtacaagg 1441 tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc aagaagaacc 1501 tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg ctgaagagaa 1561 ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa gagatcttca 1621 gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag tccttcctgg 1681 tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg gacgaggtgg 1741 cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg gacagcaccg 1801 acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag ttccggggcc 1861 acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag ctgttcatcc 1921 agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc agcggcgtgg 1981 acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa aatctgatcg 2041 cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc ctgagcctgg 2101 gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa ctgcagctga 2161 gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc gaccagtacg 2221 ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc gacatcctga 2281 gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag agatacgacg 2341 agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg cctgagaagt 2401 acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt gacggcggag 2461 ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg gacggcaccg 2521 aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg accttcgaca 2581 acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg cggcggcagg 2641 aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc ctgaccttcc 2701 gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc tggatgacca 2761 gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac aagggcgctt 2821 ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc aacgagaagg 2881 tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag ctgaccaaag 2941 tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag cagaaaaagg 3001 ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag ctgaaagagg 3061 actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt 3121 tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac aaggacttcc 3181 tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg acactgtttg 3241 aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc gacgacaaag 3301 tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc cggaagctga 3361 tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg aagtccgacg 3421 gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc tttaaagagg 3481 acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac attgccaatc 3541 tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg gtggacgagc 3601 tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg gccagagaga 3661 accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg atcgaagagg 3721 gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac acccagctgc 3781 agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac gtggaccagg 3841 aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct cagagctttc 3901 tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac cggggcaaga 3961 gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg cggcagctgc 4021 tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc gagagaggcg 4081 gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa acccggcaga 4141 tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac gacgagaatg 4201 acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg tccgatttcc 4261 ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac gcccacgacg 4321 cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag ctggaaagcg 4381 agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc aagagcgagc 4441 aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg aactttttca 4501 agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc gagacaaacg 4561 gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg cggaaagtgc 4621 tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc ggcttcagca 4681 aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag aaggactggg 4741 accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg ctggtggtgg 4801 ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg ctggggatca 4861 ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa gccaagggct 4921 acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg ttcgagctgg 4981 aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga aacgaactgg 5041 ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag aagctgaagg 5101 gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag cactacctgg 5161 acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc gacgctaatc 5221 tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga gagcaggccg 5281 agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc ttcaagtact 5341 ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg gacgccaccc 5401 tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct cagctgggag 5461 gtgacgaggg agctgataag cgcaccgccg atggttccga gttcgaaagc cccaagaaga 5521 agaggaaagt ctaaccggtc atcatcacca tcaccattga gtttaaaccc gctgatcagc 5581 ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt 5641 gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca 5701 ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga 5761 ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc 5821 ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg cgtaatcatg 5881 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc 5941 cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca cattaattgc 6001 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat 6061 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac 6121 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt 6181 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca 6241 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc 6301 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact 6361 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct 6421 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag 6481 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca 6541 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa 6601 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc 6661 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag 6721 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg 6781 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca 6841 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc 6901 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag 6961 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata 7021 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat 7081 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg 7141 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc 7201 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc 7261 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc 7321 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc 7381 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc 7441 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa 7501 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat 7561 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata 7621 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca 7681 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag 7741 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc 7801 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc 7861 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata 7921 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta 7981 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga 8041 cggatcggga gatcgatctc ccgatcccct agggtcgact ctcagtacaa tctgctctga 8101 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg 8161 cgcgag //