质粒名称: | pSIN4-EF2-ABCG2-IRES-Neo |
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出品公司: | Addgene |
目录编号: | 25983 |
存储实验室: | Ren-he Xu |
载体骨架基本信息 |
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载体名称: | pSIN-EF2-MCS-IRES-Neo |
载体出品公司: | James Thomson |
空载体大小: | 6500 bp |
完整质粒大小: | 9576 bp |
载体修饰: | -- |
载体类型: | 哺乳动物表达,慢病毒 |
筛选标记: | 新霉素(G418筛选) |
载体抗性: | 氨苄青霉素,100μg/mL |
生长温度: | 37 ℃ |
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: | ATP-binding cassette, sub-family G (WHITE), member 2, variant of isoform 2 |
物种来源: | H. sapiens (human) |
插入大小: | 1968 bp |
基因突变: | -- |
GenBank编号 | AAH92408.1 |
启动子: | -- |
标签/融合蛋白: | EF1 apha启动子(骨架上的N末端)IRES Neo(主干网上的C终端) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | EcoR I(未知是否破坏) |
3’克隆位点: | BamH I (未知是否破坏) |
5’测序引物 | EF-1a-F |
3’测序引物 | IRES-R |
文献引用方法 |
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材料和方法部分: | pSIN4-EF2-ABCG2-IRES-Neo was a gift from Ren-he Xu (Addgene plasmid # 25983 ; http://n2t.net/addgene:25983 ; RRID:Addgene_25983) |
参考文献部分: | Lack of ABCG2 expression and side population properties in human pluripotent stem cells. Zeng H, Park JW, Guo M, Lin G, Crandall L, Compton T, Wang X, Li XJ, Chen FP, Xu RH. Stem Cells. 2009 Oct . 27(10):2435-45. 10.1002/stem.192 PubMed 19670287 |
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 pSIN4-EF2-ABCG2-IRES-Neo 9576 bp ds-DNA circular SYN 16-JUN-2022 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pSIN4-EF2-ABCG2-IRES-Neo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9576) AUTHORS Zeng H, Park JW, Guo M, Lin G, Crandall L, Compton T, Wang X, Li XJ, Chen FP, Xu RH TITLE Lack of ABCG2 expression and side population properties in human pluripotent stem cells. JOURNAL Stem Cells. 2009 Oct . 27(10):2435-45. PUBMED 19670287 REFERENCE 2 (bases 1 to 9576) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 16, 2022 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9576 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(159..176) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 386..405 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 601..1404 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(664..683) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 1274..1293 /label=Neo-F /note="Neomycin resistance gene, forward primer" LTR 1607..1840 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(1991..2008) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1997..2221 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" regulatory 2264..2273 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter 2317..2421 /gene="bla" /label=AmpR promoter CDS 2422..3282 /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(2640..2659) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3453..4041 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3942..3961 /label=pBR322ori-F /note="pBR322 origin, forward primer" LTR 4200..4833 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 4880..5005 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 5498..5731 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 5916..5960 /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 6247..6364 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 6493..7671 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 6724..7662 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 7619..7639 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" misc_feature 9568..565 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" ORIGIN 1 ctcccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt 61 ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc ggaaacctgg 121 ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag gaatgcaagg 181 tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac aaacaacgtc 241 tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc tctgcggcca 301 aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag 361 ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca aggggctgaa 421 ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt gcacatgctt 481 tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt 541 tttcctttga aaaacacgat gataagcttg ccacaacccg ggataattcc tgcagccaat 601 atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 661 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 721 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 781 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 841 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 901 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 961 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 1021 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 1081 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 1141 atgcccgacg gcgatgatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 1201 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 1261 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 1321 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 1381 cgccttcttg acgagttctt ctgataatag gcggccgctc gagacctaga aaaacatgga 1441 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa 1501 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac 1561 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 1621 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 1681 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 1741 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 1801 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 1861 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gccttgacat 1921 tataatagat ttagcaggaa ttgaactagg agtggagcac acaggcaaag ttctagagct 1981 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 2041 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 2101 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 2161 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg 2221 gcttctgagg cggaaagaac cagctggggg cgcgcccctc gaggccgcca tggtcatagc 2281 tgtttgacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 2341 ttctaaatac attcaaatat gtatccgctc acgagacaat aaccctgata aatgcttcaa 2401 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt 2461 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat 2521 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag 2581 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg 2641 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata 2701 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat 2761 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc 2821 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg 2881 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac 2941 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact 3001 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa 3061 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct 3121 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc 3181 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga 3241 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac 3301 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 3361 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 3421 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 3481 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 3541 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt 3601 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 3661 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 3721 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 3781 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 3841 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 3901 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 3961 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 4021 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt ctggcctttt 4081 gcacggaccg ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct 4141 gaattatttt cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagtt 4201 ggaagggcta attcactccc aaagaagaca agatatcctt gatctgtgga tctaccacac 4261 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact 4321 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa 4381 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga 4441 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga 4501 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc 4561 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc 4621 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag 4681 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 4741 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 4801 gaccctttta gtcagtgtgg aaaatctcta gcagtggcgc ccgaacaggg acttgaaagc 4861 gaaagggaaa ccagaggagc tctctcgacg caggactcgg cttgctgaag cgcgcacggc 4921 aagaggcgag gggcggcgac tggtgagtac gccaaaaatt ttgactagcg gaggctagaa 4981 ggagagagat gggtgcgaga gcgtcagtat taagcggggg agaattagat cgcgatggga 5041 aaaaattcgg ttaaggccag ggggaaagaa aaaatataaa ttaaaacata tagtatgggc 5101 aagcagggag ctagaacgat tcgcagttaa tcctggcctg ttagaaacat cagaaggctg 5161 tagacaaata ctgggacagc tacaaccatc ccttcagaca ggatcagaag aacttagatc 5221 attatataat acagtagcaa ccctctattg tgtgcatcaa aggatagaga taaaagacac 5281 caaggaagct ttagacaaga tagaggaaga gcaaaacaaa agtaagacca ccgcacagca 5341 agcggccgct gatcttcaga cctggaggag gagatatgag ggacaattgg agaagtgaat 5401 tatataaata taaagtagta aaaattgaac cattaggagt agcacccacc aaggcaaaga 5461 gaagagtggt gcagagagaa aaaagagcag tgggaatagg agctttgttc cttgggttct 5521 tgggagcagc aggaagcact atgggcgcag cgtcaatgac gctgacggta caggccagac 5581 aattattgtc tggtatagtg cagcagcaga acaatttgct gagggctatt gaggcgcaac 5641 agcatctgtt gcaactcaca gtctggggca tcaagcagct ccaggcaaga atcctggctg 5701 tggaaagata cctaaaggat caacagctcc tggggatttg gggttgctct ggaaaactca 5761 tttgcaccac tgctgtgcct tggaatgcta gttggagtaa taaatctctg gaacagattt 5821 ggaatcacac gacctggatg gagtgggaca gagaaattaa caattacaca agcttaatac 5881 actccttaat tgaagaatcg caaaaccagc aagaaaagaa tgaacaagaa ttattggaat 5941 tagataaatg ggcaagtttg tggaattggt ttaacataac aaattggctg tggtatataa 6001 aattattcat aatgatagta ggaggcttgg taggtttaag aatagttttt gctgtacttt 6061 ctatagtgaa tagagttagg cagggatatt caccattatc gtttcagacc cacctcccaa 6121 ccccgagggg acccgacagg cccgaaggaa tagaagaaga aggtggagag agagacagag 6181 acagatccat tcgattagtg aacggatctc gacggtatcg ccacaaatgg cagtattcat 6241 ccacaatttt aaaagaaagg gggggattgg ggggtacagt gcaggggaaa gaatagtaga 6301 cataatagca acagacatac aaactaaaga attacaaaaa caaattacaa aaattcaaaa 6361 ttttcgggtt tattacaggg acagcagaga tccactttgg atcgataagc tttgcaaaga 6421 tggataaagt tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa 6481 ggagtgggaa ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc 6541 gagaagttgg ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta 6601 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg 6661 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca 6721 caggtaagtg ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc 6781 gtgccttgaa ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt 6841 ggaagtgggt gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga 6901 gttgaggcct ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc 6961 tgtctcgctg ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg 7021 ctttttttct ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg 7081 gtttttgggg ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg 7141 cggggcctgc gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct 7201 gctctggtgc ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc 7261 cggtcggcac cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc 7321 tcaaaatgga ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa 7381 agggcctttc cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc 7441 aggcacctcg attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg 7501 ttttatgcga tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg 7561 cacttgatgt aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc 7621 aagcctcaga cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaattcgc 7681 caccatgtct tccagtaatg tcgaagtttt tatcccagtg tcacaaggaa acaccaatgg 7741 cttccccgcg acagcttcca atgacctgaa ggcatttact gaaggagctg tgttaagttt 7801 tcataacatc tgctatcgag taaaactgaa gagtggcttt ctaccttgtc gaaaaccagt 7861 tgagaaagaa atattatcga atatcaatgg gatcatgaaa cctggtctca acgccatcct 7921 gggacccaca ggtggaggca aatcttcgtt attagatgtc ttagctgcaa ggaaagatcc 7981 aagtggatta tctggagatg ttctgataaa tggagcaccg cgacctgcca atttcaaatg 8041 taattcaggt tacgtggtac aagatgatgt tgtgatgggc actctgacgg tgagagaaaa 8101 cttaaagttc tcagcagctc ttcggcttgc aacaactatg acgaatcatg aaaaaaacga 8161 acggattaac agggtcattc aagagttagg tctggataaa gtggcagact ccaaggttgg 8221 aactcagttt atccgtggtg tgtctggagg agaaagaaaa aggactagta taggaatgga 8281 gcttatcact gatccttcca tcttgttctt ggatgagcct acaactggct tagactcaag 8341 cacagcaaat gctgtccttt tgctcctgaa aaggatgtct aagcagggac gaacaatcat 8401 cttctccatt catcagcctc gatattccat cttcaagttg tttgatagcc tcaccttatt 8461 ggcctcagga agacttatgt tccacgggcc tgctcaggag gccttgggat actttgaatc 8521 agctggttat cactgtgagg cctataataa ccctgcagac ttcttcttgg acatcattaa 8581 tggagattcc actgctgtgg cattaaacag agaagaagac tttaaagcca cagagatcat 8641 agagccttcc aagcaggata agccactcat agaaaaatta gcggagattt atgtcaactc 8701 ctccttctac aaagagacaa aagctgaatt acatcaactt tccgggggtg agaagaagaa 8761 gaagatcaca gtcttcaagg agatcagcta caccacctcc ttctgtcatc aactcagatg 8821 ggtttccaag cgttcattca aaaacttgct ggttaatccc caggcctcta tagctcagat 8881 cattgtcaca gtcgtactgg gactggttat aggtgccatt tactttgggc taaaaaatga 8941 ttctactgga atccagaaca gagctggggt tctcttcttc ctgacgacca accagtgttt 9001 cagcagtgtt tcagccgtgg aactctttgt ggtagagaag aagctcttca tacatgaata 9061 catcagcgga tactacagag tgtcatctta tttccttgga aaactgttat ctgatttatt 9121 acccatgagg atgttaccaa gtattatatt tacctgtata gtgtacttca tgttaggatt 9181 gaagccaaag gcagatgcct tcttcgttat gatgtttacc cttatgatgg tggcttattc 9241 agccagttcc atggcactgg ccatagcagc aggtcagagt gtggtttctg tagcaacact 9301 tctcatgacc atctgttttg tgtttatgat ggtctgttgg tcaatctcac aaccattgca 9361 tcttggctgt catggcttca gtacttcagc attccacgat atggatttac ggctttgcag 9421 cataatgaat ttttgggaca aaacttctgc ccaggactca atgcaacagg aaacaatcct 9481 tgtaactatg caacatgtac tggcgaagaa tatttggtaa ggatccgcat gcatctaggg 9541 cggccaattc cgcccctctc cccccccccc ctctcc //
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