质粒名称: | 6-His-MBP-TEV-FnCpf1 |
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出品公司: | 爱迪基因 |
目录编号: | 90094 |
存储实验室: | Feng Zhang |
载体骨架基本信息 |
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载体名称: | pET-28 |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 6643 bp |
完整质粒大小: | 10553 bp |
载体修饰: | -- |
载体类型: | 细菌表达 |
筛选标记: | -- |
高拷贝/低拷贝: | 低拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | FnCpf1 (humanized),type V CRISPR-associated protein Cpf1 (humanized),Cpf1 (humanized),Francisella tularensis subsp. novicida U112 Cpf1 (humanized) |
物种来源: | Francisella tularensis subsp. novicida U112 |
插入大小: | 3910 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | T7 |
标签/融合蛋白: |
6xHis(主干网上的N个终端)
MBP(插入N端子)
TEV站点(N端子插入)
核浆蛋白NLS(插入物上的C末端)
3xHA(插入式C端子)
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克隆方法: | 限制性内切酶 |
5' 克隆位点: | NcoI(销毁时未知) |
3' 克隆位点: | XhoI(未破坏) |
5' 测序引物 | taatacgactcactataggg |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | 6-His-MBP-TEV-FnCpf1 was a gift from Feng Zhang (Addgene plasmid # 90094 ; http://n2t.net/addgene:90094 ; RRID:Addgene_90094) |
参考文献部分: | Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F. Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038. 10.1016/j.cell.2015.09.038 PubMed 26422227 |
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 6-His-MBP-TEV-FnCpf1 10685 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Bacterial expression plasmid for protein purification. ACCESSION . VERSION . KEYWORDS 6-His-MBP-TEV-FnCpf1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10685) AUTHORS Zetsche B, Gootenberg JS, Abudayyeh OO, Slaymaker IM, Makarova KS, Essletzbichler P, Volz SE, Joung J, van der Oost J, Regev A, Koonin EV, Zhang F TITLE Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. JOURNAL Cell. 2015 Sep 23. pii: S0092-8674(15)01200-3. doi: 10.1016/j.cell.2015.09.038. PUBMED 26422227 REFERENCE 2 (bases 1 to 10685) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10685 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 532..1120 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1021..1040 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1274..1291 /label=L4440 /note="L4440 vector, forward primer" misc_feature 1306..1445 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(1371..1390) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1531..1553 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(1547..1738) /codon_start=1 /gene="rop" /product="Rop protein, which maintains plasmids at low copy number" /label=rop /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA DELYRSCLARFGDDGENL" CDS complement(2547..3629) /codon_start=1 /gene="lacI" /product="lac repressor" /label=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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" primer_bind 3591..3610 /label=LacI-R /note="LacI, reverse primer" promoter complement(3630..3707) /gene="lacI" /label=lacI promoter primer_bind 3913..3932 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 4016..4035 /label=T7 /note="T7 promoter, forward primer" promoter 4016..4034 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 4035..4059 /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)." RBS 4074..4096 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 4110..4127 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" regulatory 5082..5091 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5244..5261 /codon_start=1 /product="thrombin recognition and cleavage site" /label=thrombin site /translation="LVPRGS" CDS 5268..5288 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 9189..9236 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 9243..9323 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" CDS 9348..9371 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 9372..9416 /codon_start=1 /product="peptide tag that allows for enzymatic biotinylation" /label=AviTag(TM) /translation="GLNDIFEAQKIEWHE" CDS 9432..9449 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(9502..9520) /label=T7 Term /note="T7 terminator, reverse primer" terminator 9516..9563 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" rep_origin 9600..10055 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9687..9706) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9897..9918 /label=F1ori-F /note="F1 origin, forward primer" promoter 10081..10185 /gene="bla" /label=AmpR promoter CDS 10186..361 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(10404..10423) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac 61 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 121 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 181 gtttattgct gataaatctg gagccggtga gcgtggtagt cgcggtatca ttgcagcact 241 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 301 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 361 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 421 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 481 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 541 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 601 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 661 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 721 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 781 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 841 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 901 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 961 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 1021 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 1081 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 1141 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc 1201 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg 1261 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt 1321 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc 1381 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg 1441 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 1501 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 1561 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg 1621 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta 1681 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg 1741 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga 1801 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg 1861 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc 1921 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc 1981 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac 2041 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc 2101 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc 2161 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc gccatgccgg 2221 cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg aaggcttgag 2281 cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc 2341 gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct acgagttgca 2401 tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc caccggaagg 2461 agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc taatgagtga 2521 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 2581 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc 2641 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg 2701 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt 2761 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact 2821 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc 2881 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc 2941 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga 3001 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa 3061 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg 3121 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag 3181 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg 3241 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc 3301 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc 3361 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga 3421 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg 3481 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa 3541 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct 3601 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg 3661 cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg gatctcgacg 3721 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag 3781 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac 3841 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg 3901 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt 3961 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata 4021 cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt 4081 aactttaaga aggagatata catatgggtc atcatcatca ccatcacaaa atcgaagaag 4141 gcaaactggt tatttggatc aatggcgata aaggctataa tggtctggca gaagttggca 4201 aaaaattcga aaaagatacc ggcattaaag tgaccgttga acatccggat aaactggaag 4261 aaaaatttcc gcaggttgca gcaaccggtg atggtccgga tattatcttt tgggcacatg 4321 atcgttttgg tggttatgca cagagcggtc tgctggcaga aattacaccg gataaagcat 4381 ttcaggacaa actgtatccg tttacctggg atgcagttcg ctataacggt aaactgattg 4441 catatccgat tgcagttgaa gcactgagcc tgatctataa caaagatctg ctgccgaatc 4501 cgcctaaaac ctgggaagaa attccggcac tggataaaga actgaaagca aaaggtaaaa 4561 gcgcactgat gtttaatctg caagaaccgt attttacctg gcctctgatt gcagcagatg 4621 gtggctatgc attcaaatat gaaaacggca aatatgatat caaagacgtg ggtgttgata 4681 atgcgggtgc aaaagccggt ctgacctttc tggttgatct gattaaaaac aaacacatga 4741 acgccgatac cgattatagc attgcagaag cagcatttaa caaaggtgaa accgcaatga 4801 caattaatgg tccgtgggca tggtcaaata ttgataccag caaagtgaat tatggtgtta 4861 ccgttctgcc gacatttaaa ggtcagccga gcaaaccgtt tgttggtgtt ctgagcgcag 4921 gtattaatgc agcaagcccg aacaaagaac tggcaaaaga atttctggaa aactatctgc 4981 tgaccgatga aggtctggaa gcagtgaata aagataaacc gctgggtgca gttgcactga 5041 aaagctatga agaagaactg gttaaagatc cgcgtattgc agccaccatg gaaaatgcac 5101 agaaaggcga aattatgccg aatattccgc agatgagcgc attttggtat gccgttcgta 5161 ccgcagtgat taatgccgca tcaggtcgtc agaccgttga tgaagccctg aaagatgccc 5221 agaccggtgc aagcaccgga tccctggttc cgcgtggtag cgctagcgaa aatctgtatt 5281 ttcagggaat gagcatctac caggagttcg tcaacaagta ttcactgagt aagacactgc 5341 ggttcgagct gatcccacag ggcaagacac tggagaacat caaggcccga ggcctgattc 5401 tggacgatga gaagcgggca aaagactata agaaagccaa gcagatcatt gataaatacc 5461 accagttctt tatcgaggaa attctgagct ccgtgtgcat cagtgaggat ctgctgcaga 5521 attactcaga cgtgtacttc aagctgaaga agagcgacga tgacaacctg cagaaggact 5581 tcaagtccgc caaggacacc atcaagaaac agattagcga gtacatcaag gactccgaaa 5641 agtttaaaaa tctgttcaac cagaatctga tcgatgctaa gaaaggccag gagtccgacc 5701 tgatcctgtg gctgaaacag tctaaggaca atgggattga actgttcaag gctaactccg 5761 atatcactga tattgacgag gcactggaaa tcatcaagag cttcaaggga tggaccacat 5821 actttaaagg cttccacgag aaccgcaaga acgtgtactc cagcaacgac attcctacct 5881 ccatcatcta ccgaatcgtc gatgacaatc tgccaaagtt cctggagaac aaggccaaat 5941 atgaatctct gaaggacaaa gctcccgagg caattaatta cgaacagatc aagaaagatc 6001 tggctgagga actgacattc gatatcgact ataagactag cgaggtgaac cagagggtct 6061 tttccctgga cgaggtgttt gaaatcgcca atttcaacaa ttacctgaac cagtccggca 6121 ttactaaatt caataccatc attggcggga agtttgtgaa cggggagaat accaagcgca 6181 agggaattaa cgaatacatc aatctgtata gccagcagat caacgacaaa actctgaaga 6241 aatacaagat gtctgtgctg ttcaaacaga tcctgagtga taccgagtcc aagtcttttg 6301 tcattgataa actggaagat gactcagacg tggtcactac catgcagagc ttttatgagc 6361 agatcgccgc tttcaagaca gtggaggaaa aatctattaa ggaaactctg agtctgctgt 6421 tcgatgacct gaaagcccag aagctggacc tgagtaagat ctacttcaaa aacgataaga 6481 gtctgacaga cctgtcacag caggtgtttg atgactattc cgtgattggg accgccgtcc 6541 tggagtacat tacacagcag atcgctccaa agaacctgga taatccctct aagaaagagc 6601 aggaactgat cgctaagaaa accgagaagg caaaatatct gagtctggaa acaattaagc 6661 tggcactgga ggagttcaac aagcacaggg atattgacaa acagtgccgc tttgaggaaa 6721 tcctggccaa cttcgcagcc atccccatga tttttgatga gatcgcccag aacaaagaca 6781 atctggctca gatcagtatt aagtaccaga accagggcaa gaaagacctg ctgcaggctt 6841 cagcagaaga tgacgtgaaa gccatcaagg atctgctgga ccagaccaac aatctgctgc 6901 acaagctgaa aatcttccat attagtcagt cagaggataa ggctaatatc ctggataaag 6961 acgaacactt ctacctggtg ttcgaggaat gttacttcga gctggcaaac attgtccccc 7021 tgtataacaa gattaggaac tacatcacac agaagcctta ctctgacgag aagtttaaac 7081 tgaacttcga aaatagtacc ctggccaacg ggtgggataa gaacaaggag cctgacaaca 7141 cagctatcct gttcatcaag gatgacaagt actatctggg agtgatgaat aagaaaaaca 7201 ataagatctt cgatgacaaa gccattaagg agaacaaagg ggaaggatac aagaaaatcg 7261 tgtataagct gctgcccggc gcaaataaga tgctgcctaa ggtgttcttc agcgccaaga 7321 gtatcaaatt ctacaaccca tccgaggaca tcctgcggat tagaaatcac tcaacacata 7381 ctaagaacgg gagcccccag aagggatatg agaaatttga gttcaacatc gaggattgca 7441 ggaagtttat tgacttctac aagcagagca tctccaaaca ccctgaatgg aaggattttg 7501 gcttccggtt ttccgacaca cagagatata actctatcga cgagttctac cgcgaggtgg 7561 aaaatcaggg gtataagctg acttttgaga acatttctga aagttacatc gacagcgtgg 7621 tcaatcaggg aaagctgtac ctgttccaga tctataacaa agatttttca gcatacagca 7681 agggcagacc aaacctgcat acactgtact ggaaggccct gttcgatgag aggaatctgc 7741 aggacgtggt ctataaactg aacggagagg ccgaactgtt ttaccggaag cagtctattc 7801 ctaagaaaat cactcaccca gctaaggagg ccatcgctaa caagaacaag gacaatccta 7861 agaaagagag cgtgttcgaa tacgatctga ttaaggacaa gcggttcacc gaagataagt 7921 tctttttcca ttgtccaatc accattaact tcaagtcaag cggcgctaac aagttcaacg 7981 acgagatcaa tctgctgctg aaggaaaaag caaacgatgt gcacatcctg agcattgacc 8041 gaggagagcg gcatctggcc tactataccc tggtggatgg caaagggaat atcattaagc 8101 aggatacatt caacatcatt ggcaatgacc ggatgaaaac caactaccac gataaactgg 8161 ctgcaatcga gaaggataga gactcagcta ggaaggactg gaagaaaatc aacaacatta 8221 aggagatgaa ggaaggctat ctgagccagg tggtccatga gattgcaaag ctggtcatcg 8281 aatacaatgc cattgtggtg ttcgaggatc tgaacttcgg ctttaagagg gggcgcttta 8341 aggtggaaaa acaggtctat cagaagctgg agaaaatgct gatcgaaaag ctgaattacc 8401 tggtgtttaa agataacgag ttcgacaaga ccggaggcgt cctgagagcc taccagctga 8461 cagctccctt tgaaactttc aagaaaatgg gaaaacagac aggcatcatc tactatgtgc 8521 cagccggatt cacttccaag atctgccccg tgaccggctt tgtcaaccag ctgtacccta 8581 aatatgagtc agtgagcaag tcccaggaat ttttcagcaa gttcgataag atctgttata 8641 atctggacaa ggggtacttc gagttttcct tcgattacaa gaacttcggc gacaaggccg 8701 ctaaggggaa atggaccatt gcctccttcg gatctcgcct gatcaacttt cgaaattccg 8761 ataaaaacca caattgggac actagggagg tgtacccaac caaggagctg gaaaagctgc 8821 tgaaagacta ctctatcgag tatggacatg gcgaatgcat caaggcagcc atctgtggcg 8881 agagtgataa gaaatttttc gccaagctga cctcagtgct gaatacaatc ctgcagatgc 8941 ggaactcaaa gaccgggaca gaactggact atctgattag ccccgtggct gatgtcaacg 9001 gaaacttctt cgacagcaga caggcaccca aaaatatgcc tcaggatgca gacgccaacg 9061 gggcctacca catcgggctg aagggactga tgctgctggg ccggatcaag aacaatcagg 9121 aggggaagaa gctgaacctg gtcattaaga acgaggaata cttcgagttt gtccagaata 9181 gaaataacaa aaggccggcg gccacgaaaa aggccggcca ggcaaaaaag aaaaagggat 9241 cctacccata cgatgttcca gattacgctt atccctacga cgtgcctgat tatgcatacc 9301 catatgatgt ccccgactat gcctaaagcc tcgagggtgg tggtagcgat tataaagatg 9361 atgatgataa aggcctgaac gatatctttg aagcccagaa aattgaatgg cacgagtaaa 9421 agcttgtcga gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg 9481 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta 9541 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga 9601 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc 9661 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac 9721 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag 9781 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc 9841 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg 9901 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata 9961 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa 10021 cgcgaatttt aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg 10081 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga 10141 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat 10201 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca 10261 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc 10321 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca 10381 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg 10441 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca 10501 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata 10561 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag 10621 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg 10681 gagct //
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