质粒名称: | pEP4 E02S CK2M EN2L |
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出品公司: | 爱迪基因 |
目录编号: | 20924 |
存储实验室: | James Thomson |
载体骨架基本信息 |
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载体名称: | pCEP4 |
载体出品公司: | Invitrogen |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | 11600 bp |
载体修饰: | -- |
载体类型: | -- |
筛选标记: | 新霉素(G418) |
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: | Oct4 and Sox2; Nanog and Lin28; Klf4 and Myc |
物种来源: | -- |
插入大小: | 4900 bp |
基因突变: | -- |
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: | AgeI(未破坏) |
3' 克隆位点: | NotI (未破坏) |
5' 测序引物 | CMV-F |
3' 测序引物 | SV40pA-R |
文献引用方法 |
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材料和方法部分: | pEP4 E02S CK2M EN2L was a gift from James Thomson (Addgene plasmid # 20924 ; http://n2t.net/addgene:20924 ; RRID:Addgene_20924) |
参考文献部分: | Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences. Yu J, Hu K, Smuga-Otto K, Tian S, Stewart R, Slukvin II, Thomson JA. Science. 2009 Mar 26. 10.1126/science.1172482 PubMed 19325077 |
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 pEP4 E02S CK2M EN2L 19949 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION used in the derivation of human iPS cells using non-integrating episomal vectors; expresses Oct4 and Sox2; Klf4 and Myc; Nanog and Lin28. ACCESSION . VERSION . KEYWORDS pEP4 E02S CK2M EN2L SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 19949) AUTHORS Yu J, Hu K, Smuga-Otto K, Tian S, Stewart R, Slukvin II, Thomson JA TITLE Human Induced Pluripotent Stem Cells Free of Vector and Transgene Sequences JOURNAL Science. 2009 Mar 26. PUBMED 19325077 REFERENCE 2 (bases 1 to 19949) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..19949 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 13..1175 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 244..1166 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" misc_feature 2385..2958 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 2408..2960 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" polyA_signal 3977..4111 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" enhancer 4618..4997 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 4998..5201 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 5152..5172 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" misc_feature 6713..7286 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 6736..7288 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" CDS 7348..8667 /codon_start=1 /product="human c-Myc proto-oncogene" /label=c-Myc /translation="MPLNVSFTNRNYDLDYDSVQPYFYCDEEENFYQQQQQSELQPPAP SEDIWKKFELLPTPPLSPSRRSGLCSPSYVAVTPFSLRGDNDGGGGSFSTADQLEMVTE LLGGDMVNQSFICDPDDETFIKNIIIQDCMWSGFSAAAKLVSEKLASYQAARKDSGSPN PARGHSVCSTSSLYLQDLSAAASECIDPSVVFPYPLNDSSSPKSCASQDSSAFSPSSDS LLSSTESSPQGSPEPLVLHEETPPTTSSDSEEEQEDEEEIDVVSVEKRQAPGKRSESGS PSAGGHSKPPHSPLVLKRCHVSTHQHNYAAPPSTRKDYPAAKRVKLDSVRVLRQISNNR KCTSPRSSDTEENVKRRTHNVLERQRRNELKRSFFALRDQIPELENNEKAPKVVILKKA TAYILSVQAEEQKLISEEDLLRKRREQLKHKLEQLRNSCA" primer_bind complement(8724..8741) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 8730..8954 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" promoter 9175..10337 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 9406..10328 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" misc_feature 11381..11954 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 11404..11956 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" polyA_signal 12650..12784 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin 13515..15304 /label=oriP /note="Epstein-Barr virus oriP replication origin (Yates et al., 2000)" CDS complement(15606..17531) /codon_start=1 /product="Epstein-Barr nuclear antigen 1, also known as EBNA-1" /label=EBNA1 /note="crucial for latent viral infection, and for episomal amplification of vectors containing the oriP origin (Okita et al., 2013)" /translation="MSDEGPGTGPGNGLGEKGDTSGPEGSGGSGPQRRGGDNHGRGRGR GRGRGGGRPGAPGGSGSGPRHRDGVRRPQKRPSCIGCKGTHGGTGAGAGAGGAGAGGAG AGGGAGAGGGAGGAGGAGGAGAGGGAGAGGGAGGAGGAGAGGGAGAGGGAGGAGAGGGA GGAGGAGAGGGAGAGGGAGGAGAGGGAGGAGGAGAGGGAGAGGAGGAGGAGAGGAGAGG GAGGAGGAGAGGAGAGGAGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGGAGAG GGAGGAGAGGAGGAGAGGAGGAGAGGAGGAGAGGGAGAGGAGAGGGGRGRGGSGGRGRG GSGGRGRGGSGGRRGRGRERARGGSRERARGRGRGRGEKRPRSPSSQSSSSGSPPRRPP PGRRPFFHPVGEADYFEYHQEGGPDGEPDVPPGAIEQGPADDPGEGPSTGPRGQGDGGR RKKGGWFGKHRGQGGSNPKFENIAEGLRALLARSHVERTTDEGTWVAGVFVYGGSKTSL YNLRRGTALAIPQCRLTPLSRLPFGMAPGPGPQPGPLRESIVCYFMVFLQTHIFAEVLK DAIKDLVMTKPAPTCNIRVTVCSFDDGVDLPPWFPPMVEGAAAEGDDGDDGDEGGDGDE GEEGQE" primer_bind complement(17970..17988) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 18056..18160 /gene="bla" /label=AmpR promoter CDS 18161..19021 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(18379..18398) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 19192..19780 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 19681..19700 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctagcttcgt gaggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 61 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta 121 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 181 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 241 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc 301 gtgccttgaa ttacttccac ctggctccag tacgtgattc ttgatcccga gctggagcca 361 ggggcgggcc ttgcgcttta ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg 421 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg 481 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag 541 atagtcttgt aaatgcgggc caggatctgc acactggtat ttcggttttt gggcccgcgg 601 ccggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc 661 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc 721 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg 781 cgtgagcgga aagatggccg cttcccggcc ctgctccagg gggctcaaaa tggaggacgc 841 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct 901 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt 961 tctggagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt 1021 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct 1081 cgttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg 1141 ttcaaagttt ttttcttcca tttcaggtgt cgtgaacacg tggtcgcggc caattctgca 1201 gtcgagctca agcttcgaac tagtgattat ggcgggacac ctggcttcgg atttcgcctt 1261 ctcgccccct ccaggtggtg gaggtgatgg gccagggggg ccggagccgg gctgggttga 1321 tcctcggacc tggctaagct tccaaggccc tcctggaggg ccaggaatcg ggccgggggt 1381 tgggccaggc tctgaggtgt gggggattcc cccatgcccc ccgccgtatg agttctgtgg 1441 ggggatggcg tactgtgggc cccaggttgg agtggggcta gtgccccaag gcggcttgga 1501 gacctctcag cctgagggtg aagcangagt cggggtggag agcaactccg atggggcctc 1561 cccggagccc tgcaccgtca cccctggtgc cgtgaagctg gagaaggaga agctggagca 1621 aaacccggag gagtcccagg acatcaaagc tctgcagaaa gaactcgagc aatttgccaa 1681 gctcctgaag cagaagagga tcaccctggg atatacacag gccgatgtgg ggctcaccct 1741 gggggttcta tttgggaagg tattcagcca aacgaccatc tgccgctttg aggctctgca 1801 gcttagcttc aagaacatgt gtaagctgcg gcccttgctg cagaagtggg tggaggaagc 1861 tgacaacaat gaaaatcttc aggagatatg caaagcagaa accctcgtgc aggcccgaaa 1921 gagaaagcga accagtatcg agaaccgagt gagaggcaac ctggagaatt tgttcctgca 1981 gtgcccgaaa cccacactgc agcagatcag ccacatcgcc cagcagcttg ggctcgagaa 2041 ggatgtggtc cgagtgtggt tctgtaaccg gcgccagaag ggcaagcgat caagcagcga 2101 ctatgcacaa cgagaggatt ttgaggctgc tgggtctcct ttctcagggg gaccagtgtc 2161 ctttcctctg gccccagggc cccattttgg taccccaggc tatgggagcc ctcacttcac 2221 tgcactgtac tcctcggtcc ctttccctga gggggaagcc tttccccctg tctccgtcac 2281 cactctgggc tctcccatgc attcaaactg aggtgcctgc cctctagttt acgcgtgggg 2341 gcggccggcc gctttctcga ggccatcgat gtttaaacga tccgcccctc tccctccccc 2401 ccccctaacg ttactggccg aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg 2461 ttattttcca ccatattgcc gtcttttggc aatgtgaggg cccggaaacc tggccctgtc 2521 ttcttgacga gcattcctag gggtctttcc cctctcgcca aaggaatgca aggtctgttg 2581 aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg 2641 accctttgca ggcagcggaa ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca 2701 cgtgtataag atacacctgc aaaggcggca caaccccagt gccacgttgt gagttggata 2761 gttgtggaaa gagtcaaatg gctctcctca agcgtattca acaaggggct gaaggatgcc 2821 cagaaggtac cccattgtat gggatctgat ctggggcctc ggtacacatg ctttacatgt 2881 gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc acggggacgt ggttttcctt 2941 tgaaaaacac gatgataata tggccacacc gcgaattaat tcgattccgc atgtacaaca 3001 tgatggagac ggagctgaag ccgccgggcc cgcagcaaac ttcggggggc ggcggcggca 3061 actccaccgc ggcggcggcc ggcggcaacc agaaaaacag cccggaccgc gtcaagcggc 3121 ccatgaatgc cttcatggtg tggtcccgcg ggcagcggcg caagatggcc caggagaacc 3181 ccaagatgca caactcggag atcagcaagc gcctgggcgc cgagtggaaa cttttgtcgg 3241 agacggagaa gcggccgttc atcgacgagg ctaagcggct gcgagcgctg cacatgaagg 3301 agcacccgga ttataaatac cggccccggc ggaaaaccaa gacgctcatg aagaaggata 3361 agtacacgct gcccggcggg ctgctggccc ccggcggcaa tagcatggcg agcggggtcg 3421 gggtgggcgc cggcctgggc gcgggcgtga accagcgcat ggacagttac gcgcacatga 3481 acggctggag caacggcagc tacagcatga tgcaggacca gctgggctac ccgcagcacc 3541 cgggcctcaa tgcgcacggc gcagcgcaga tgcagcccat gcaccgctac gacgtgagcg 3601 ccctgcagta caactccatg accagctcgc agacctacat gaacggctcg cccacctaca 3661 gcatgtccta ctcgcagcag ggcacccctg gcatggctct tggctccatg ggttcggtgg 3721 tcaagtccga ggccagctcc agcccccctg tggttacctc ttcctcccac tccagggcgc 3781 cctgccaggc cggggacctc cgggacatga tcagcatgta tctccccggc gccgaggtgc 3841 cggaacccgc cgcccccagc agacttcaca tgtcccagca ctaccagagc ggcccggtgc 3901 ccggcacggc cattaacggc acactgcccc tctcacacat gtgagggccg gaaatcacta 3961 gctagttcga aggatcgatc cagacatgat aagatacatt gatgagtttg gacaaaccac 4021 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt 4081 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt 4141 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg 4201 tatggctgat tatgatccgg ctgcctcgcg cgtttcggtg atgacggtga aaacctctga 4261 cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa 4321 gcccgtcagg cgtcagcggg tgttggcggg tgtcggggcg cagccatgag gtcgaccaat 4381 tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc 4441 gcagaactgg taggtatgga agatcgatct atacattgaa tcaatattgg caattagcca 4501 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt 4561 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac 4621 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 4681 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 4741 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 4801 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 4861 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 4921 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 4981 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 5041 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 5101 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 5161 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga 5221 tccgctagga tctcgagctc aagcttcgaa ttcgcgatcg ccatggctgt cagcgacgcg 5281 ctgctcccat ctttctccac gttcgcgtct ggcccggcgg gaagggagaa gacactgcgt 5341 caagcaggtg ccccgaataa ccgctggcgg gaggagctct cccacatgaa gcgacttccc 5401 ccagtgcttc ccggccgccc ctatgacctg gcggcggcga ccgtggccac agacctggag 5461 agcggcggag ccggtgcggc ttgcggcggt agcaacctgg cgcccctacc tcggagagag 5521 accgaggagt tcaacgatct cctggacctg gactttattc tctccaattc gctgacccat 5581 cctccggagt cagtggccgc caccgtgtcc tcgtcagcgt cagcctcctc ttcgtcgtcg 5641 ccgtcgagca gcggccctgc cagcgcgccc tccacctgca gcttcaccta tccgatccgg 5701 gccgggaacg acccgggcgt ggcgccgggc ggcacgggcg gaggcctcct ctatggcagg 5761 gagtccgctc cccctccgac ggctcccttc aacctggcgg acatcaacga cgtgagcccc 5821 tcgggcggct tcatggccga gctcctgcgg ccagaattgg acccggtgta cattccgccg 5881 cagcagccgc agccgccagg tggcgggctg atgggcaagt tcgtgctgaa ggcgtcgctg 5941 agcgcccctg gcagcgagta cggcagcccg tcggtcatca gcgtcagcaa aggcagccct 6001 gacggcagcc acccggtggt ggtggcgccc tacaacggcg ggccgccgcg cacgtgcccc 6061 aagatcaagc aggaggcggt ctcttcgtgc acccacttgg gcgctggacc ccctctcagc 6121 aatggccacc ggccggctgc acacgacttc cccctggggc ggcagctccc cagcaggact 6181 accccgaccc tgggtcttga ggaagtgctg agcagcaggg actgtcaccc tgccctgccg 6241 cttcctcccg gcttccatcc ccacccgggg cccaattacc catccttcct gcccgatcag 6301 atgcagccgc aagtcccgcc gctccattac caagagctca tgccacccgg ttcctgcatg 6361 ccagaggagc ccaagccaaa gaggggaaga cgatcgtggc cccggaaaag gaccgccacc 6421 cacacttgtg attacgcggg ctgcggcaaa acctacacaa agagttccca tctcaaggca 6481 cacctgcgaa cccacacagg tgagaaacct taccactgtg actgggacgg ctgtggatgg 6541 aaattcgccc gctcagatga actgaccagg cactaccgta aacacacggg gcaccgcccg 6601 ttccagtgcc aaaaatgcga ccgagcattt tccaggtcgg accacctcgc cttacacatg 6661 aagaggcatt tttaaatccc agacagtgga tatgacccgt ttaaacgatc cgcccctctc 6721 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg 6781 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg 6841 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag 6901 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt 6961 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc 7021 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga 7081 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga 7141 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct 7201 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg 7261 ttttcctttg aaaaacacga tgataatatg gccacaccgc gatcgcgaat tctgcagtcg 7321 agctcaagct tcgaattcgc gatcgccatg cccctcaacg ttagcttcac caacaggaac 7381 tatgacctcg actacgactc ggtgcagccg tatttctact gcgacgagga ggagaacttc 7441 taccagcagc agcagcagag cgagctgcag cccccggcgc ccagcgagga tatctggaag 7501 aaattcgagc tgctgcccac cccgcccctg tcccctagcc gccgctccgg gctctgctcg 7561 ccctcctacg ttgcggtcac acccttctcc cttcggggag acaacgacgg cggtggcggg 7621 agcttctcca cggccgacca gctggagatg gtgaccgagc tgctgggagg agacatggtg 7681 aaccagagtt tcatctgcga cccggacgac gagaccttca tcaaaaacat catcatccag 7741 gactgtatgt ggagcggctt ctcggccgcc gccaagctcg tctcagagaa gctggcctcc 7801 taccaggctg cgcgcaaaga cagcggcagc ccgaaccccg cccgcggcca cagcgtctgc 7861 tccacctcca gcttgtacct gcaggatctg agcgccgccg cctcagagtg catcgacccc 7921 tcggtggtct tcccctaccc tctcaacgac agcagctcgc ccaagtcctg cgcctcgcaa 7981 gactccagcg ccttctctcc gtcctcggat tctctgctct cctcgacgga gtcctccccg 8041 cagggcagcc ccgagcccct ggtgctccat gaggagacac cgcccaccac cagcagcgac 8101 tctgaggagg aacaagaaga tgaggaagaa atcgatgttg tttctgtgga aaagaggcag 8161 gctcctggca aaaggtcaga gtctggatca ccttctgctg gaggccacag caaacctcct 8221 cacagcccac tggtcctcaa gaggtgccac gtctccacac atcagcacaa ctacgcagcg 8281 cctccctcca ctcggaagga ctatcctgct gccaagaggg tcaagttgga cagtgtcaga 8341 gtcctgagac agatcagcaa caaccgaaaa tgcaccagcc ccaggtcctc ggacaccgag 8401 gagaatgtca agaggcgaac acacaacgtc ttggagcgcc agaggaggaa cgagctaaaa 8461 cggagctttt ttgccctgcg tgaccagatc ccggagttgg aaaacaatga aaaggccccc 8521 aaggtagtta tccttaaaaa agccacagca tacatcctgt ccgtccaagc agaggagcaa 8581 aagctcattt ctgaagagga cttgttgcgg aaacgacgag aacagttgaa acacaaactt 8641 gaacagctac ggaactcttg tgcgtaagtt tggccgcccc cacgcgtaaa ctagttcgaa 8701 ggatcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt 8761 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta 8821 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg 8881 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc 8941 ggtgggctct atggcttctg aggcggaaag aaccagctgg gggcgcgccc ctcgagatct 9001 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca 9061 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg 9121 gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagcttc gtgaggctcc 9181 ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg 9241 gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga aagtgatgtc 9301 gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag tgcagtagtc 9361 gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag tgccgtgtgt 9421 ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg aattacttcc 9481 acctggctcc agtacgtgat tcttgatccc gagctggagc caggggcggg ccttgcgctt 9541 taggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg 9601 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt 9661 aaaatttttg atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg 9721 gccaggatct gcacactggt atttcggttt ttgggcccgc ggccggcgac ggggcccgtg 9781 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac 9841 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg 9901 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc 9961 cgcttcccgg ccctgctcca gggggctcaa aatggaggac gcggcgctcg ggagagcggg 10021 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg 10081 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctggagc ttttggagta 10141 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg 10201 tggagactga agttaggcca gcttggcact tgatgtaatt ctcgttggaa tttgcccttt 10261 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc 10321 catttcaggt gtcgtgaaca cgtggtcgcg gccaattctg cagtcgagct caagcttcga 10381 actagggccg cgactctagt gattcatgag tgtggatcca gcttgtcccc aaagcttgcc 10441 ttgctttgaa gcatccgact gtaaagaatc ttcacctatg cctgtgattt gtgggcctga 10501 agaaaactat ccatccttgc aaatgtcttc tgctgagatg cctcacacgg agactgtctc 10561 tcctcttcct tcctccatgg atctgcttat tcaggacagc cctgattctt ccaccagtcc 10621 caaaggcaaa caacccactt ctgcagagaa gagtgtcgca aaaaaggaag acaaggtccc 10681 ggtcaagaaa cagaagacca gaactgtgtt ctcttccacc cagctgtgtg tactcaatga 10741 tagatttcag agacagaaat acctcagcct ccagcagatg caagaactct ccaacatcct 10801 gaacctcagc tacaaacagg tgaagacctg gttccagaac cagagaatga aatctaagag 10861 gtggcagaaa aacaactggc cgaagaatag caatggtgtg acgcagaagg cctcagcacc 10921 tacctacccc agcctttact cttcctacca ccagggatgc ctggtgaacc cgactgggaa 10981 ccttccaatg tggagcaacc agacctggaa caattcaacc tggagcaacc agacccagaa 11041 catccagtcc tggagcaacc actcctggaa cactcagacc tggtgcaccc aatcctggaa 11101 caatcaggcc tggaacagtc ccttctataa ctgtggagag gaatctctgc agtcctgcat 11161 gcagttccag ccaaattctc ctgccagtga cttggaggct gccttggaag ctgctgggga 11221 aggccttaat gtaatacagc agaccactag gtattttagt actccacaaa ccatggattt 11281 attcctaaac tactccatga acatgcaacc tgaagacgtg tgaagatgaa tcgaattccc 11341 gcggccgctt tctcgaggcc atcgatgttt aaacgatccg cccctctccc tccccccccc 11401 ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc tatatgttat 11461 tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc cctgtcttct 11521 tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt ctgttgaatg 11581 tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct gtagcgaccc 11641 tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa aagccacgtg 11701 tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt tggatagttg 11761 tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag gatgcccaga 11821 aggtacccca ttgtatggga tctgatctgg ggcctcggta cacatgcttt acatgtgttt 11881 agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt ttcctttgaa 11941 aaacacgatg ataatatggc cacaccgcga tcgcgaattc gattatcgcc atgggctccg 12001 tgtccaacca gcagtttgca ggtggctgcg ccaaggcggc agaagaggcg cccgaggagg 12061 cgccggagga cgcggcccgg gcggcggacg agcctcagct gctgcacggt gcgggcatct 12121 gtaagtggtt caacgtgcgc atggggttcg gcttcctgtc catgaccgcc cgcgccgggg 12181 tcgcgctcga ccccccagtg gatgtctttg tgcaccagag taagctgcac atggaagggt 12241 tccggagctt gaaggagggt gaggcagtgg agttcacctt taagaagtca gccaagggtc 12301 tggaatccat ccgtgtcacc ggacctggtg gagtattctg tattgggagt gagaggcggc 12361 caaaaggaaa gagcatgcag aagcgcagat caaaaggaga caggtgctac aactgtggag 12421 gtctagatca tcatgccaag gaatgcaagc tgccacccca gcccaagaag tgccacttct 12481 gccagagcat cagccatatg gtagcctcat gtccgctgaa ggcccagcag ggccctagtg 12541 cacagggaaa gccaacctac tttcgagagg aagaagaaga aatccacagc cctaccctgc 12601 tcccggaggc acagaattga gtttaaacna atcactagct agttcgaagg atccagacat 12661 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt 12721 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca 12781 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt 12841 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc cggctgcctc 12901 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca 12961 gcttgtctgt aagcggatgc cgggagcaga caagcccgtc aggcgtcagc gggtgttggc 13021 gggtgtcggg gcgcagccat gaggtcgatc gactctagct agaggatcga tgccccgccc 13081 cggacgaact aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa 13141 tcccttcagt tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg 13201 gggaccaaac acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac 13261 atttgccaac actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac 13321 acaacattgc ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat 13381 gtacctccca ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc 13441 tgtgtagcta ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc 13501 ccttgttaac cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac 13561 taaccctaat tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt 13621 agtaaaaggg tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt 13681 tacatggggt caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg 13741 aagatcaagg agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc 13801 gtttagctaa tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa 13861 aacaaggttt caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat 13921 tgtgctatga caccaatata accctcacaa accccttggg caataaatac tagtgtagga 13981 atgaaacatt ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag 14041 actggatgtc catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata 14101 tgatactggg gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt 14161 tacactctat ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg 14221 ttgtctctaa cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa 14281 gacaagtggc cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac 14341 gccgccctgc ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc 14401 gctaaccact gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac 14461 ctgcataagt aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa 14521 ttacacacac ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt 14581 cgctgagagc acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga 14641 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg 14701 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg 14761 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg 14821 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg 14881 tagtatatgc tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc 14941 atatgctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc 15001 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt 15061 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc 15121 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc 15181 atatgctatc ctcatgcata tacagtcagc atatgatacc cagtagtaga gtgggagtgc 15241 tatcctttgc atatgccgcc acctcccaag ggggcgtgaa ttttcgctgc ttgtcctttt 15301 cctgctggtt gctcccattc ttaggtgaat ttaaggaggc caggctaaag ccgtcgcatg 15361 tctgattgct caccaggtaa atgtcgctaa tgttttccaa cgcgagaagg tgttgagcgc 15421 ggagctgagt gacgtgacaa catgggtatg cccaattgcc ccatgttggg aggacgaaaa 15481 tggtgacaag acagatggcc agaaatacac caacagcacg catgatgtct actggggatt 15541 tattctttag tgcgggggaa tacacggctt ttaatacgat tgagggcgtc tcctaacaag 15601 ttacatcact cctgcccttc ctcaccctca tctccatcac ctccttcatc tccgtcatct 15661 ccgtcatcac cctccgcggc agccccttcc accataggtg gaaaccaggg aggcaaatct 15721 actccatcgt caaagctgca cacagtcacc ctgatattgc aggtaggagc gggctttgtc 15781 ataacaaggt ccttaatcgc atccttcaaa acctcagcaa atatatgagt ttgtaaaaag 15841 accatgaaat aacagacaat ggactccctt agcgggccag gttgtgggcc gggtccaggg 15901 gccattccaa aggggagacg actcaatggt gtaagacgac attgtggaat agcaagggca 15961 gttcctcgcc ttaggttgta aagggaggtc ttactacctc catatacgaa cacaccggcg 16021 acccaagttc cttcgtcggt agtcctttct acgtgactcc tagccaggag agctcttaaa 16081 ccttctgcaa tgttctcaaa tttcgggttg gaacctcctt gaccacgatg ctttccaaac 16141 caccctcctt ttttgcgcct gcctccatca ccctgacccc ggggtccagt gcttgggcct 16201 tctcctgggt catctgcggg gccctgctct atcgctcccg ggggcacgtc aggctcacca 16261 tctgggccac cttcttggtg gtattcaaaa taatcggctt cccctacagg gtggaaaaat 16321 ggccttctac ctggaggggg cctgcgcggt ggagacccgg atgatgatga ctgactactg 16381 ggactcctgg gcctcttttc tccacgtcca cgacctctcc ccctggctct ttcacgactt 16441 ccccccctgg ctctttcacg tcctctaccc cggcggcctc cactacctcc tcgaccccgg 16501 cctccactac ctcctcgacc ccggcctcca ctgcctcctc gaccccggcc tccacctcct 16561 gctcctgccc ctcctgctcc tgcccctcct cctgctcctg cccctcctgc ccctcctgct 16621 cctgcccctc ctgcccctcc tgctcctgcc cctcctgccc ctcctgctcc tgcccctcct 16681 gcccctcctc ctgctcctgc ccctcctgcc cctcctcctg ctcctgcccc tcctgcccct 16741 cctgctcctg cccctcctgc ccctcctgct cctgcccctc ctgcccctcc tgctcctgcc 16801 cctcctgctc ctgcccctcc tgctcctgcc cctcctgctc ctgcccctcc tgcccctcct 16861 gcccctcctc ctgctcctgc ccctcctgct cctgcccctc ctgcccctcc tgcccctcct 16921 gctcctgccc ctcctcctgc tcctgcccct cctgcccctc ctgcccctcc tcctgctcct 16981 gcccctcctg cccctcctcc tgctcctgcc cctcctcctg ctcctgcccc tcctgcccct 17041 cctgcccctc ctcctgctcc tgcccctcct gcccctcctc ctgctcctgc ccctcctcct 17101 gctcctgccc ctcctgcccc tcctgcccct cctcctgctc ctgcccctcc tcctgctcct 17161 gcccctcctg cccctcctgc ccctcctgcc cctcctcctg ctcctgcccc tcctcctgct 17221 cctgcccctc ctgctcctgc ccctcccgct cctgctcctg ctcctgttcc accgtgggtc 17281 cctttgcagc caatgcaact tggacgtttt tggggtctcc ggacaccatc tctatgtctt 17341 ggccctgatc ctgagccgcc cggggctcct ggtcttccgc ctcctcgtcc tcgtcctctt 17401 ccccgtcctc gtccatggtt atcaccccct cttctttgag gtccactgcc gccggagcct 17461 tctggtccag atgtgtctcc cttctctcct aggccatttc caggtcctgt acctggcccc 17521 tcgtcagaca tgattcacac taaaagagat caatagacat ctttattaga cgacgctcag 17581 tgaatacagg gagtgcagac tcctgccccc tccaacagcc cccccaccct catccccttc 17641 atggtcgctg tcagacagat ccaggtctga aaattcccca tcctccgaac catcctcgtc 17701 ctcatcacca attactcgca gcccggaaaa ctcccgctga acatcctcaa gatttgcgtc 17761 ctgagcctca agccaggcct caaattcctc gtcccccttt ttgctggacg gtagggatgg 17821 ggattctcgg gacccctcct cttcctcttc aaggtcacca gacagagatg ctactggggc 17881 aacggaagaa aagctgggtg cggcctgtga ggatcagctt atcgatgata agctgtcaaa 17941 catgagaatt aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa 18001 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg 18061 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata 18121 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg 18181 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac 18241 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact 18301 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat 18361 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga 18421 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac 18481 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat 18541 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac 18601 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct 18661 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac 18721 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga 18781 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg 18841 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact 18901 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac 18961 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta 19021 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt 19081 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga 19141 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc 19201 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt 19261 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc 19321 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc 19381 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg 19441 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg 19501 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga 19561 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc 19621 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg 19681 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg 19741 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt 19801 tttacggttc ctggcctttt gctggccttg aagctgtccc tgatggtcgt catctacctg 19861 cctggacagc atggcctgca acgcgggcat cccgatgccg ccggaagcga gaagaatcat 19921 aatggggaag gccatccagc ctcgcgtcg //
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