质粒名称: | pRRLsin-SV40 T antigen-IRES-mCherry |
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出品公司: | Addgene |
目录编号: | 58993 |
存储实验室: | Snorri Thorgeirsson |
载体骨架基本信息 |
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载体名称: | pRRLsin.cPPT.PGK/GFP.WPRE |
载体出品公司: | -- |
空载体大小: | 6110 bp |
完整质粒大小: | 11405 bp |
载体修饰: | 位于nt 924的NotI站点被删除 |
载体类型: | 哺乳细胞表达,慢病毒 |
筛选标记: | mCherry 红色荧光 |
载体抗性: | 氨苄霉素 |
生长温度: | 30 ℃ |
宿主菌: | Stbl3 |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | CMV enhancer-chicken beta actin promoter |
物种来源: | -- |
插入大小: | 1730 bp |
基因突变: | MCS 中的 EcoRV 站点转换为 BamHI 站点 |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5’克隆位点: | SalI(克隆期间销毁) |
3’克隆位点: | BamHI(未破坏) |
5’测序引物 | -- |
3’测序引物 | pCAG-F |
文献引用方法 |
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材料和方法部分: | pRRLsin-SV40 T antigen-IRES-mCherry was a gift from Snorri Thorgeirsson (Addgene plasmid # 58993 ; http://n2t.net/addgene:58993 ; RRID:Addgene_58993) |
参考文献部分: | Modeling pathogenesis of primary liver cancer in lineage-specific mouse cell types. Holczbauer A, Factor VM, Andersen JB, Marquardt JU, Kleiner DE, Raggi C, Kitade M, Seo D, Akita H, Durkin ME, Thorgeirsson SS. Gastroenterology. 2013 Jul;145(1):221-31. doi: 10.1053/j.gastro.2013.03.013. Epub 2013 Mar 19. 10.1053/j.gastro.2013.03.013 PubMed 23523670 |
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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 pRRLsin-SV40 T antigen-IRES-mCherry 11418 bp ds-DNA circular SYN 08-SEP-2021 DEFINITION lentiviral expression of SV40 large T antigen and mCherry red fluorescent reporter. ACCESSION . VERSION . KEYWORDS pRRLsin-SV40 T antigen-IRES-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11418) AUTHORS Holczbauer A, Factor VM, Andersen JB, Marquardt JU, Kleiner DE, Raggi C, Kitade M, Seo D, Akita H, Durkin ME, Thorgeirsson SS TITLE Modeling pathogenesis of primary liver cancer in lineage-specific mouse cell types. JOURNAL Gastroenterology. 2013 Jul;145(1):221-31. doi: 10.1053/j.gastro.2013.03.013. Epub 2013 Mar 19. PUBMED 23523670 REFERENCE 2 (bases 1 to 11418) AUTHORS . TITLE Direct Submission JOURNAL Exported Sep 8, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11418 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 8..234 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 235..415 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 462..587 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1085..1318 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1503..1547 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 1814..1931 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 1953..2332 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 2611..3627 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 3551..3573 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 3635..3654 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 3726..5852 /codon_start=1 /product="large T antigen from SV40 (simian virus 40)" /label=large T antigen /translation="MDKVLNREESLQLMDLLGLERSAWGNIPLMRKAYLKKCKEFHPDK GGDEEKMKKMNTLYKKMEDGVKYAHQPDFGGFWDATEIPTYGTDEWEQWWNAFNEENLF CSEEMPSSDDEATADSQHSTPPKKKRKVEDPKDFPSELLSFLSHAVFSNRTLACFAIYT TKEKAALLYKKIMEKYSVTFISRHNSYNHNILFFLTPHRHRVSAINNYAQKLCTFSFLI CKGVNKEYLMYSALTRDPFSVIEESLPGGLKEHDFNPEEAEETKQVSWKLVTEYAMETK CDDVLLLLGMYLEFQYSFEMCLKCIKKEQPSHYKYHEKHYANAAIFADSKNQKTICQQA VDTVLAKKRVDSLQLTREQMLTNRFNDLLDRMDIMFGSTGSADIEEWMAGVAWLHCLLP KMDSVVYDFLKCMVYNIPKKRYWLFKGPIDSGKTTLAAALLELCGGKALNVNLPLDRLN FELGVAIDQFLVVFEDVKGTGGESRDLPSGQGINNLDNLRDYLDGSVKVNLEKKHLNKR TQIFPPGIVTMNEYSVPKTLQARFVKQIDFRPKDYLKHCLERSEFLLEKRIIQSGIALL LMLIWYRPVAEFAQSIQSRIVEWKERLDKEFSLSVYQKMKFNVAMGIGVLDWLRNSDDD DEDSQENADKNEDGGEKNMEDSGHETGIDSQSQGSFQAPQSSQSVHDHNQPYHICRGFT CFKKPPTPPPEPET" misc_feature 5921..6507 /label=IRES2 /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(6088..6105) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 6315..6334 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 6508..7218 /codon_start=1 /product="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /label=mCherry /note="mammalian codon-optimized" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind complement(6653..6671) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 6921..6940 /label=mCherry-F /note="mCherry, forward primer" primer_bind complement(7238..7254) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7238..7254) /label=pBluescriptSK /note="For pBluescript vector" misc_feature 7269..7857 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7322..7342) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7740..7751) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7944..8177 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 8249..8370 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(8286..8305) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 8340..8359 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 8410..8545 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8472..8491 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(8565..8584) /label=T7 /note="T7 promoter, forward primer" promoter complement(8566..8584) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(8594..8611) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(8594..8610) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(8603..8625) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 8752..9207 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8839..8858) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 9049..9070 /label=F1ori-F /note="F1 origin, forward primer" promoter 9233..9337 /gene="bla" /label=AmpR promoter CDS 9338..10198 /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 complement(9556..9575) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 10369..10957 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 10858..10877 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 11111..11128 /label=L4440 /note="L4440 vector, forward primer" protein_bind 11245..11266 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 11281..11311 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11319..11335 /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)." primer_bind 11324..11346 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 11343..11359 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 11343..11359 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 11378..11398 /label=T3 /note="T3 promoter, forward primer" promoter 11380..11398 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" ORIGIN 1 agcttaatgt agtcttatgc aatactcttg tagtcttgca acatggtaac gatgagttag 61 caacatgcct tacaaggaga gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg 121 tacgatcgtg ccttattagg aaggcaacag acgggtctga catggattgg acgaaccact 181 gaattgccgc attgcagaga tattgtattt aagtgcctag ctcgatacat aaacgggtct 241 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 301 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 361 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 421 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 481 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 541 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 601 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 661 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 721 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 781 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 841 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 901 aaagtaagac caccgcacag caagcggcct ggccgctgat cttcagacct ggaggaggag 961 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat 1021 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg 1081 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt 1141 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca 1201 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca 1261 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg 1321 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt 1381 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag 1441 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag 1501 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta 1561 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag 1621 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac 1681 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag 1741 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac 1801 ggtatcggtt aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa 1861 tagtagacat aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa 1921 ttcaaaattt tatcgatcac gagactagcc tcgacattga ttattgacta gttattaata 1981 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact 2041 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat 2101 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta 2161 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc 2221 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg 2281 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt 2341 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta 2401 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggnsgcgcg 2461 ccrggsgggg sggggsgggg sgrggggsgg ggsggggsgr ggsggrgagg tgcggcggca 2521 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg 2581 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg 2641 ccccgctccg ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca 2701 caggtgagcg ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga 2761 cggcttgttt cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg 2821 tgcgggggga gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc 2881 ggctccgcgc tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc 2941 cgcagtgtgc gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc 3001 gaggggaaca aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc 3061 gcgtcggtcg ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg 3121 gcttcgggtg cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg 3181 tggcggcagg tgggggtgcc gggcggggcg gggccgcctc gggccggggr gggctcgggg 3241 grggggcgcg gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat 3301 tgccttttat ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg 3361 gagccgaaat ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc 3421 ggcgccggca ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc 3481 cttctccctc tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg 3541 ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc tgctaaccat 3601 gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat tgtgctgtct 3661 catcattttg gcaaagaatt cctcgaggat gaggatcccg cgaccggtcg accagctttg 3721 caaagatgga taaagtttta aacagagagg aatctttgca gctaatggac cttctaggtc 3781 ttgaaaggag tgcctggggg aatattcctc tgatgagaaa ggcatattta aaaaaatgca 3841 aggagtttca tcctgataaa ggaggagatg aagaaaaaat gaagaaaatg aatactctgt 3901 acaagaaaat ggaagatgga gtaaaatatg ctcatcaacc tgactttgga ggcttctggg 3961 atgcaactga gattccaacc tatggaactg atgaatggga gcagtggtgg aatgccttta 4021 atgaggaaaa cctgttttgc tcagaagaaa tgccatctag tgatgatgag gctactgctg 4081 actctcaaca ttctactcct ccaaaaaaga agagaaaggt agaagacccc aaggactttc 4141 cttcagaatt gctaagtttt ttgagtcatg ctgtgtttag taatagaact cttgcttgct 4201 ttgctattta caccacaaag gaaaaagctg cactgctata caagaaaatt atggaaaaat 4261 attctgtaac ctttataagt aggcataaca gttataatca taacatactg ttttttctta 4321 ctccacacag gcatagagtg tctgctatta ataactatgc tcaaaaattg tgtaccttta 4381 gctttttaat ttgtaaaggg gttaataagg aatatttgat gtatagtgcc ttgactagag 4441 atccattttc tgttattgag gaaagtttgc caggtgggtt aaaggagcat gattttaatc 4501 cagaagaagc agaggaaact aaacaagtgt cctggaagct tgtaacagag tatgcaatgg 4561 aaacaaaatg tgatgatgtg ttgttattgc ttgggatgta cttggaattt cagtacagtt 4621 ttgaaatgtg tttaaaatgt attaaaaaag aacagcccag ccactataag taccatgaaa 4681 agcattatgc aaatgctgct atatttgctg acagcaaaaa ccaaaaaacc atatgccaac 4741 aggctgttga tactgtttta gctaaaaagc gggttgatag cctacaatta actagagaac 4801 aaatgttaac aaacagattt aatgatcttt tggataggat ggatataatg tttggttcta 4861 caggctctgc tgacatagaa gaatggatgg ctggagttgc ttggctacac tgtttgttgc 4921 ccaaaatgga ttcagtggtg tatgactttt taaaatgcat ggtgtacaac attcctaaaa 4981 aaagatactg gctgtttaaa ggaccaattg atagtggtaa aactacatta gcagctgctt 5041 tgcttgaatt atgtgggggg aaagctttaa atgttaattt gcccttggac aggctgaact 5101 ttgagctagg agtagctatt gaccagtttt tagtagtttt tgaggatgta aagggcactg 5161 gaggggagtc cagagatttg ccttcaggtc agggaattaa taacctggac aatttaaggg 5221 attatttgga tggcagtgtt aaggtaaact tagaaaagaa acacctaaat aaaagaactc 5281 aaatatttcc ccctggaata gtcaccatga atgagtacag tgtgcctaaa acactgcagg 5341 ccagatttgt aaaacaaata gattttaggc ccaaagatta tttaaagcat tgcctggaac 5401 gcagtgagtt tttgttagaa aagagaataa ttcaaagtgg cattgctttg cttcttatgt 5461 taatttggta cagacctgtg gctgagtttg ctcaaagtat tcagagcaga attgtggagt 5521 ggaaagagag attggacaaa gagtttagtt tgtcagtgta tcaaaaaatg aagtttaatg 5581 tggctatggg aattggagtt ttagattggc taagaaacag tgatgatgat gatgaagaca 5641 gccaggaaaa tgctgataaa aatgaagatg gtggggagaa gaacatggaa gactcagggc 5701 atgaaacagg cattgattca cagtcccaag gctcatttca ggcccctcag tcctcacagt 5761 ctgttcatga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 5821 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacggg 5881 gatcctcatc gcggccgcta cgtaaattcc gccccccccc cccctctccc tccccccccc 5941 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat 6001 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct 6061 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg 6121 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc 6181 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg 6241 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg 6301 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga 6361 aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt acatgtgttt 6421 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa 6481 aaacacgatg ataatatggc cacaaccatg gtgagcaagg gcgaggagga taacatggcc 6541 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag 6601 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg 6661 aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg 6721 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc 6781 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc 6841 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc 6901 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc 6961 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag 7021 ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc 7081 gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag 7141 gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg 7201 gacgagctgt acaagtaaga attcctgcag cccgggggat ccactagttc tagagcggcc 7261 gcgtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 7321 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 7381 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 7441 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 7501 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc 7561 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 7621 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat 7681 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 7741 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 7801 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcctg 7861 gaattcgagc tcggtacctt taagaccaat gacttacaag gcagctgtag atcttagcca 7921 ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa gacaagatct 7981 gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg 8041 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt 8101 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt 8161 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc 8221 aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata atggttacaa 8281 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg 8341 tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat cccgccccta 8401 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga 8461 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag 8521 tagtgaggag gcttttttgg aggcctaggg acgtacccaa ttcgccctat agtgagtcgt 8581 attacgcgcg ctcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta 8641 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg 8701 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg gacgcgccct 8761 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg 8821 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg 8881 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac 8941 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct 9001 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt 9061 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt 9121 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt 9181 ttaacaaaat attaacgctt acaatttagg tggcactttt cggggaaatg tgcgcggaac 9241 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 9301 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 9361 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 9421 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 9481 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 9541 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 9601 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 9661 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 9721 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc 9781 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 9841 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 9901 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 9961 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 10021 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 10081 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 10141 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact 10201 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa 10261 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt 10321 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt 10381 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 10441 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 10501 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt 10561 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 10621 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 10681 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 10741 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 10801 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 10861 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 10921 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 10981 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga 11041 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac 11101 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc 11161 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 11221 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc 11281 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca 11341 cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga 11401 acaaaagctg gagctgca //
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