质粒名称: | pLenti-CMV-MCS-GFP-SV-puro |
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出品公司: | 爱迪基因 |
目录编号: | 73582 |
存储实验室: | Paul Odgren |
载体骨架基本信息 |
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载体名称: | pLenti-CMV-MCs-SV-puro |
载体出品公司: | gift from Dr. Hong Zhang, University of Massachusetts Medical School |
载体抗性: | 氨苄青霉素 |
空载体大小: | 8829 bp |
载体修饰: | GFP insert |
载体类型: | 慢病毒表达 |
筛选标记: | 嘌呤霉素 |
生长条件: | 37 ℃ |
插入基因信息 |
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标签/融合蛋白: | GFP tag(主干上的C终端) |
克隆方法: | 限制性内切酶 |
5' 测序引物 | GAGCACCCAGTCCGCCCTG |
文献引用方法 |
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材料和方法部分: | pLenti-CMV-MCS-GFP-SV-puro was a gift from Paul Odgren (Addgene plasmid # 73582 ; http://n2t.net/addgene:73582 ; RRID:Addgene_73582) |
参考文献部分: | Studies of OC-STAMP in Osteoclast Fusion: A New Knockout Mouse Model, Rescue of Cell Fusion, and Transmembrane Topology. Witwicka H, Hwang SY, Reyes-Gutierrez P, Jia H, Odgren PE, Donahue LR, Birnbaum MJ, Odgren PR. PLoS One. 2015 Jun 4;10(6):e0128275. doi: 10.1371/journal.pone.0128275. eCollection 2015. PONE-D-14-50827 [pii] PubMed 26042409 |
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载体基本信息
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载体简介
载体序列
LOCUS pLenti-CMV-MCS-GFP-SV-puro 8455 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Third generation lentiviral vector expressing GFP and puromycin. ACCESSION . VERSION . KEYWORDS pLenti-CMV-MCS-GFP-SV-puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8455) AUTHORS Witwicka H, Hwang SY, Reyes-Gutierrez P, Jia H, Odgren PE, Donahue LR, Birnbaum MJ, Odgren PR TITLE Studies of OC-STAMP in Osteoclast Fusion: A New Knockout Mouse Model, Rescue of Cell Fusion, and Transmembrane Topology. JOURNAL PLoS One. 2015 Jun 4;10(6):e0128275. doi: 10.1371/journal.pone.0128275. eCollection 2015. PUBMED 26042409 REFERENCE 2 (bases 1 to 8455) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8455 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 113..134 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 149..179 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 187..203 /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 192..214 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 211..227 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 211..227 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 246..266 /label=T3 /note="T3 promoter, forward primer" promoter 248..266 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" promoter 294..520 /label=RSV promoter /note="Rous sarcoma virus enhancer/promoter" LTR 521..701 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 748..873 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1366..1599 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 2095..2211 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2234..2537 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2538..2741 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2692..2712 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2738..2762 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2818..3537 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2863..2884) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3124..3143) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3471..3492 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3582..3602) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3587..3916 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3767..3902 /label=SV40 ori /note="SV40 origin of replication" CDS 3926..4525 /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(3926..3945) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 4422..4442 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 4592..5180 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4645..4665) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5063..5074) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 5267..5500 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" polyA_signal 5572..5693 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5609..5628) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5663..5682 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 5733..5868 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5888..5907) /label=T7 /note="T7 promoter, forward primer" promoter complement(5889..5907) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5917..5934) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5917..5933) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5926..5948) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 6075..6530 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6162..6181) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6372..6393 /label=F1ori-F /note="F1 origin, forward primer" promoter 6556..6660 /gene="bla" /label=AmpR promoter CDS 6661..7521 /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(6879..6898) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7692..8280 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8181..8200 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8434..8451 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca 61 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga 121 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt 181 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca 241 agcgcgcaat taaccctcac taaagggaac aaaagctgga gctgcaagct taatgtagtc 301 ttatgcaata ctcttgtagt cttgcaacat ggtaacgatg agttagcaac atgccttaca 361 aggagagaaa aagcaccgtg catgccgatt ggtggaagta aggtggtacg atcgtgcctt 421 attaggaagg caacagacgg gtctgacatg gattggacga accactgaat tgccgcattg 481 cagagatatt gtatttaagt gcctagctcg atacataaac gggtctctct ggttagacca 541 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag 601 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag 661 atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc gaacagggac 721 ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct tgctgaagcg 781 cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt gactagcgga 841 ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag aattagatcg 901 cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt aaaacatata 961 gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt agaaacatca 1021 gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg atcagaagaa 1081 cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag gatagagata 1141 aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag taagaccacc 1201 gcacagcaag cggccgctga tcttcagacc tggaggagga gatatgaggg acaattggag 1261 aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag cacccaccaa 1321 ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag ctttgttcct 1381 tgggttcttg ggagcagcag gaagcactat gggcgcagcg tcaatgacgc tgacggtaca 1441 ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga gggctattga 1501 ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc aggcaagaat 1561 cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg gttgctctgg 1621 aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata aatctctgga 1681 acagatttgg aatcacacga cctggatgga gtgggacaga gaaattaaca attacacaag 1741 cttaatacac tccttaattg aagaatcgca aaaccagcaa gaaaagaatg aacaagaatt 1801 attggaatta gataaatggg caagtttgtg gaattggttt aacataacaa attggctgtg 1861 gtatataaaa ttattcataa tgatagtagg aggcttggta ggtttaagaa tagtttttgc 1921 tgtactttct atagtgaata gagttaggca gggatattca ccattatcgt ttcagaccca 1981 cctcccaacc ccgaggggac ccgacaggcc cgaaggaata gaagaagaag gtggagagag 2041 agacagagac agatccattc gattagtgaa cggatctcga cggtatcggt taacttttaa 2101 aagaaaaggg gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac 2161 agacatacaa actaaagaat tacaaaaaca aattacaaaa ttcaaaattt tatcgataag 2221 cttgggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 2281 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 2341 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 2401 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 2461 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 2521 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 2581 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 2641 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 2701 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga 2761 tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccga ctctagaatg 2821 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc 2881 gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc 2941 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc 3001 gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag 3061 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc 3121 aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg 3181 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag 3241 ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc 3301 atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac 3361 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac 3421 ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg 3481 ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaacgg 3541 gatccggact agttaacgcg tcgaccagtg gtcgaccctg tggaatgtgt gtcagttagg 3601 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta 3661 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat 3721 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac 3781 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga 3841 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg 3901 cctaggcttt tgcaaaaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac 3961 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc 4021 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact 4081 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc 4141 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg 4201 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct 4261 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc 4321 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga 4381 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga 4441 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg 4501 catgacccgc aagcccggtg cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag 4561 gagcgcacga ccccatgcat cgccagtcga caatcaacct ctggattaca aaatttgtga 4621 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 4681 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 4741 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 4801 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 4861 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 4921 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 4981 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg 5041 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 5101 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 5161 cctttgggcc gcctccccgc ctggaattcg agctcggtac ctttaagacc aatgacttac 5221 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt 5281 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag 5341 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc 5401 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga 5461 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt 5521 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt 5581 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca 5641 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc 5701 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg 5761 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct 5821 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc 5881 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg 5941 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 6001 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 6061 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 6121 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 6181 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 6241 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 6301 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 6361 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 6421 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 6481 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact 6541 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg 6601 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt 6661 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct 6721 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca 6781 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc 6841 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc 6901 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg 6961 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta 7021 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc 7081 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt 7141 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg 7201 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct 7261 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc 7321 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct 7381 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac 7441 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc 7501 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat 7561 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg 7621 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc 7681 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa 7741 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag 7801 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta 7861 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta 7921 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag 7981 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg 8041 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg 8101 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag 8161 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc 8221 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa 8281 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg 8341 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 8401 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaag //