质粒名称: | pU6-sgRNA EF1Alpha-puro-T2A-BFP |
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出品公司: | Addgene |
目录编号: | 60955 |
存储实验室: | Jonathan Weissman |
载体骨架基本信息 |
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载体名称: | pSICO衍生物 |
载体出品公司: | Addgene |
空载体大小: | 8200 bp |
完整质粒大小: | 8888 bp |
载体修饰: | -- |
载体类型: | 哺乳动物表达,慢病毒,CRISPR |
筛选标记: | -- |
载体抗性: | 氨苄青霉素,100μg/mL |
生长温度: | 37 ℃ |
宿主菌: | Stbl3 |
拷贝数: | 低拷贝 |
插入基因信息 |
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插入基因名称: | sgGFP-NT2 |
物种来源: | -- |
插入大小: | 113 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | -- |
克隆方法: | -- |
5’克隆位点: | -- |
3’克隆位点: | -- |
5’测序引物 | -- |
3’测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pU6-sgRNA EF1Alpha-puro-T2A-BFP was a gift from Jonathan Weissman (Addgene plasmid # 60955 ; http://n2t.net/addgene:60955 ; RRID:Addgene_60955) |
参考文献部分: | Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M, Weissman JS. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct 9. 10.1016/j.cell.2014.09.029 PubMed 25307932 |
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 pU6-sgRNA EF1Alpha-puro-T2A-BFP 8877 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Parental vector for the CRISPRi/a libraries. Expresses an sgRNA from the U6 promoter and a puromycin resistance cassette and BFP from the EF1Alpha promoter. ACCESSION . VERSION . KEYWORDS pU6-sgRNA EF1Alpha-puro-T2A-BFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8877) AUTHORS Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M, Weissman JS TITLE Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. JOURNAL Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct 9. PUBMED 25307932 REFERENCE 2 (bases 1 to 8877) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..8877 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1684..1917 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2102..2146 /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 2444..2561 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" protein_bind 2814..2847 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." primer_bind 2851..2871 /label=mU6-F /note="Mouse U6 promoter, forward primer" promoter 3116..3327 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" intron 3328..4266 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 4223..4243 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 4287..4883 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4287..4306) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4783..4803 /label=Puro-F /note="Puromycin resistance gene, forward primer" CDS 4893..4946 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 4959..5657 /codon_start=1 /product="monomeric blue fluorescent protein" /label=TagBFP /note="mammalian codon-optimized" /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD LPSKLGHKLN" protein_bind 5677..5710 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5766..6354 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5819..5839) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6237..6248) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(6357..6373) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6358..6374) /label=pBluescriptKS /note="For pBluescript vector" LTR 6883..7063 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" rep_origin complement(7125..7713) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7205..7224) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7884..8744) /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 8507..8526 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8745..8849) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga 1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt 1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg 1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc 1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa 1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa 1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg 1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg 1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga 2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga 2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa 2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg 2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc 2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga 2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact 2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg 2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa 2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca 2581 gcagagatcc agtttggtta gtaccgggcc cgctctagag atccgacgcg ccatctctag 2641 gcccgcgccg gccccctcgc acggacttgt gggagaagct cggctactcc cctgccccgg 2701 ttaatttgca tataatattt cctagtaact atagaggctt aatgtgcgat aaaagacaga 2761 taatctgttc tttttaatac tagctacatt ttacatgata ggcttggatt tctataactt 2821 cgtatagcat acattatacg aagttataaa cagcacaaaa ggaaactcac cctaactgta 2881 aagtaattgt gtgttttgag actataagta tcccttggag aaccaccttg ttggaccagg 2941 atgggcacca cccgtttaag agctaagctg gaaacagcat agcaagttta aataaggcta 3001 gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttctcg agtactagga 3061 tccattaggc ggccgcgtgg ataaccgtat taccgccatg catgtgcccg tcagtgggca 3121 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt 3181 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt 3241 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt 3301 cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc 3361 ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg 3421 attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa 3481 ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg 3541 cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa 3601 aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc 3661 caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg 3721 tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg 3781 gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc 3841 ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg 3901 cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg 3961 ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac 4021 tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgcgctt ttggagtacg 4081 tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg 4141 gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt 4201 gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca 4261 tttcaggtgt cgtgagctag cccaccatga ccgagtacaa gcccacggtg cgcctcgcca 4321 cccgcgacga cgtcccccgg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg 4381 ccacgcgcca caccgtcgac ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac 4441 tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg 4501 cggtggcggt ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg 4561 gcccgcgcat ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc 4621 tcctggcgcc gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc 4681 ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg 4741 agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg 4801 agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt 4861 gcatgacccg caagcccggt gccggatcgg gagagggcag aggaagtctg ctaacatgcg 4921 gtgacgtcga ggagaatcct ggcccaccgg tcgccaccag cgagctgatt aaggagaaca 4981 tgcacatgaa gctgtacatg gagggcaccg tggacaacca tcacttcaag tgcacatccg 5041 agggcgaagg caagccctac gagggcaccc agaccatgag aatcaaggtg gtcgagggcg 5101 gccctctccc cttcgccttc gacatcctgg ctactagctt cctctacggc agcaagacct 5161 tcatcaacca cacccagggc atccccgact tcttcaagca gtccttccct gagggcttca 5221 catgggagag agtcaccaca tacgaagacg ggggcgtgct gaccgctacc caggacacca 5281 gcctccagga cggctgcctc atctacaacg tcaagatcag aggggtgaac ttcacatcca 5341 acggccctgt gatgcagaag aaaacactcg gctgggaggc cttcaccgag acgctgtacc 5401 ccgctgacgg cggcctggaa ggcagaaacg acatggccct gaagctcgtg ggcgggagcc 5461 atctgatcgc aaacatcaag accacatata gatccaagaa acccgctaag aacctcaaga 5521 tgcctggcgt ctactatgtg gactacagac tggaaagaat caaggaggcc aacaacgaga 5581 cctacgtcga gcagcacgag gtggcagtgg ccagatactg cgacctccct agcaaactgg 5641 ggcacaagct taattaagaa ttcgtcgagg gacctaataa cttcgtatag catacattat 5701 acgaagttat acatgtttaa gggttccggt tccactaggt acaattcgat atcaagctta 5761 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg 5821 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt 5881 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg 5941 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc 6001 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc 6061 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc 6121 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc 6181 tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg 6241 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc 6301 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat 6361 accgtcgacc tcgatcgaga cctagaaaaa catggagcaa tcacaagtag caatacagca 6421 gctaccaatg ctgattgtgc ctggctagaa gcacaagagg aggaggaggt gggttttcca 6481 gtcacacctc aggtaccttt aagaccaatg acttacaagg cagctgtaga tcttagccac 6541 tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag acaagatatc 6601 cttgatctgt ggatctacca cacacaaggc tacttccctg attggcagaa ctacacacca 6661 gggccaggga tcagatatcc actgaccttt ggatggtgct acaagctagt accagttgag 6721 caagagaagg tagaagaagc caatgaagga gagaacaccc gcttgttaca ccctgtgagc 6781 ctgcatggga tggatgaccc ggagagagaa gtattagagt ggaggtttga cagccgccta 6841 gcatttcatc acatggcccg agagctgcat ccggactgta ctgggtctct ctggttagac 6901 cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa 6961 agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag 7021 agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagcatgtg agcaaaaggc 7081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 7141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 7201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 7261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 7321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 7381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 7441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 7501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 7561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 7621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 7681 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 7741 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 7801 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 7861 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 7921 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 7981 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 8041 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 8101 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 8161 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 8221 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 8281 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 8341 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 8401 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 8461 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 8521 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 8581 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 8641 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 8701 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 8761 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 8821 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac //