质粒名称: | pBABE-neo largeTcDNA |
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出品公司: | 爱迪基因 |
目录编号: | 1780 |
存储实验室: | Bob Weinberg |
载体骨架基本信息 |
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载体名称: | pBABE-neo |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 5330 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳动物表达,逆转录病毒 |
筛选标记: | 新霉素(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | large T antigen,large T cDNA,pBNT,LTc |
物种来源: | simian virus |
插入大小: | 2200 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: | BamHI(未破坏) |
3' 克隆位点: | BamHI(未破坏) |
5' 测序引物 | pBABE 5' |
3' 测序引物 | pBABE-3 |
文献引用方法 |
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材料和方法部分: | pBABE-neo largeTcDNA was a gift from Bob Weinberg (Addgene plasmid # 1780 ; http://n2t.net/addgene:1780 ; RRID:Addgene_1780) |
参考文献部分: | Enumeration of the simian virus 40 early region elements necessary for human cell transformation. Hahn WC, Dessain SK, Brooks MW, King JE, Elenbaas B, Sabatini DM, DeCaprio JA, Weinberg RA. Mol Cell Biol 2002 Apr;22(7):2111-23. 10.1128/MCB.22.7.2111-2123.2002 PubMed 11884599 |
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载体基本信息
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载体序列
LOCUS pBABE-neo largeTcDNA 7464 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Retroviral expression of Large T Antigen. Used to create immortalized cell lines.. ACCESSION . VERSION . KEYWORDS pBABE-neo largeTcDNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7464) AUTHORS Hahn WC, Dessain SK, Brooks MW, King JE, Elenbaas B, Sabatini DM, DeCaprio JA, Weinberg RA TITLE Enumeration of the simian virus 40 early region elements necessary for human cell transformation. JOURNAL Mol Cell Biol 2002 Apr;22(7):2111-23. PUBMED 11884599 REFERENCE 2 (bases 1 to 7464) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7464 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 6..475 /note="long terminal repeat from Moloney murine leukemia virus" misc_feature 540..739 /label=MMLV Psi /note="packaging signal of Moloney murine leukemia virus (MMLV)" CDS 940..1356 /label=gag (truncated) /note="truncated Moloney murine leukemia virus (MMLV) gag gene lacking the start codon" primer_bind 1286..1308 /label=pLXSN 5' /note="Murine stem cell virus, forward primer. Also called MSCV" primer_bind 1324..1340 /label=pBABE 5' /note="Psi packaging signal, forward primer for pBABE vectors" CDS 1394..3520 /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" primer_bind complement(3601..3621) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3606..3935 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3786..3921 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3848..3867 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4292..5086 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(4346..4365) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4956..4975 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 5609..5628 /label=pBR322ori-F /note="pBR322 origin, forward primer" LTR 5890..6359 /note="long terminal repeat from Moloney murine leukemia virus" CDS complement(6474..7334) /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 7097..7116 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7335..7439) /gene="bla" /label=AmpR promoter ORIGIN 1 gcagcctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg 61 gccaagaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc 121 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt 181 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt 241 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc 301 tcaataaaag agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc 361 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg 421 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg 481 gggctcgtcc gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggca 541 agctggccag caacttatct gtgtctgtcc gattgtctag tgtctatgac tgattttatg 601 cgcctgcgtc ggtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact 661 gacgagttct gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt 721 ttttgtggcc cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg 781 gttctggtag gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg 841 gtttggaacc gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt 901 ctctgtctga ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc 961 ttaagtttga ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta 1021 gatgtcaaga agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc 1081 ggatggccgc gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc 1141 ttttcacctg gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc 1201 ttggcttttg acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct 1261 cttcctccat ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc 1321 tccctttatc cagccctcac tccttctcta ggcgccggcc ggatcccgcg accggtcgac 1381 cagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct 1441 tctaggtctt gaaaggagtg cctgggggaa tattcctctg atgagaaagg catatttaaa 1501 aaaatgcaag gagtttcatc ctgataaagg aggagatgaa gaaaaaatga agaaaatgaa 1561 tactctgtac aagaaaatgg aagatggagt aaaatatgct catcaacctg actttggagg 1621 cttctgggat gcaactgaga ttccaaccta tggaactgat gaatgggagc agtggtggaa 1681 tgcctttaat gaggaaaacc tgttttgctc agaagaaatg ccatctagtg atgatgaggc 1741 tactgctgac tctcaacatt ctactcctcc aaaaaagaag agaaaggtag aagaccccaa 1801 ggactttcct tcagaattgc taagtttttt gagtcatgct gtgtttagta atagaactct 1861 tgcttgcttt gctatttaca ccacaaagga aaaagctgca ctgctataca agaaaattat 1921 ggaaaaatat tctgtaacct ttataagtag gcataacagt tataatcata acatactgtt 1981 ttttcttact ccacacaggc atagagtgtc tgctattaat aactatgctc aaaaattgtg 2041 tacctttagc tttttaattt gtaaaggggt taataaggaa tatttgatgt atagtgcctt 2101 gactagagat ccattttctg ttattgagga aagtttgcca ggtgggttaa aggagcatga 2161 ttttaatcca gaagaagcag aggaaactaa acaagtgtcc tggaagcttg taacagagta 2221 tgcaatggaa acaaaatgtg atgatgtgtt gttattgctt gggatgtact tggaatttca 2281 gtacagtttt gaaatgtgtt taaaatgtat taaaaaagaa cagcccagcc actataagta 2341 ccatgaaaag cattatgcaa atgctgctat atttgctgac agcaaaaacc aaaaaaccat 2401 atgccaacag gctgttgata ctgttttagc taaaaagcgg gttgatagcc tacaattaac 2461 tagagaacaa atgttaacaa acagatttaa tgatcttttg gataggatgg atataatgtt 2521 tggttctaca ggctctgctg acatagaaga atggatggct ggagttgctt ggctacactg 2581 tttgttgccc aaaatggatt cagtggtgta tgacttttta aaatgcatgg tgtacaacat 2641 tcctaaaaaa agatactggc tgtttaaagg accaattgat agtggtaaaa ctacattagc 2701 agctgctttg cttgaattat gtggggggaa agctttaaat gttaatttgc ccttggacag 2761 gctgaacttt gagctaggag tagctattga ccagttttta gtagtttttg aggatgtaaa 2821 gggcactgga ggggagtcca gagatttgcc ttcaggtcag ggaattaata acctggacaa 2881 tttaagggat tatttggatg gcagtgttaa ggtaaactta gaaaagaaac acctaaataa 2941 aagaactcaa atatttcccc ctggaatagt caccatgaat gagtacagtg tgcctaaaac 3001 actgcaggcc agatttgtaa aacaaataga ttttaggccc aaagattatt taaagcattg 3061 cctggaacgc agtgagtttt tgttagaaaa gagaataatt caaagtggca ttgctttgct 3121 tcttatgtta atttggtaca gacctgtggc tgagtttgct caaagtattc agagcagaat 3181 tgtggagtgg aaagagagat tggacaaaga gtttagtttg tcagtgtatc aaaaaatgaa 3241 gtttaatgtg gctatgggaa ttggagtttt agattggcta agaaacagtg atgatgatga 3301 tgaagacagc caggaaaatg ctgataaaaa tgaagatggt ggggagaaga acatggaaga 3361 ctcagggcat gaaacaggca ttgattcaca gtcccaaggc tcatttcagg cccctcagtc 3421 ctcacagtct gttcatgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt 3481 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg 3541 ttaacgggga tcccagtgtg gtggtacgta ggaattcgcc agcacagtgg tcgaccctgt 3601 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 3661 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag 3721 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc 3781 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa 3841 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt 3901 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tcacgctgcc gcaagcactc 3961 agggcgcaag ggctgctaaa ggaagcggaa cacgtagaaa gccagtccgc agaaacggtg 4021 ctgaccccgg atgaatgtca gctactgggc tatctggaca agggaaaacg caagcgcaaa 4081 gagaaagcag gtagcttgca gtgggcttac atggcgatag ctagactggg cggttttatg 4141 gacagcaagc gaaccggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg 4201 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc 4261 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 4321 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 4381 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 4441 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 4501 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 4561 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 4621 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 4681 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 4741 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 4801 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 4861 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 4921 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 4981 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 5041 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 5101 ttcgcatcga tgcatcgatg cggcgggtgt ggtggttacg cgtaatctgc tgcttgcaaa 5161 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 5221 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 5281 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 5341 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 5401 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 5461 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 5521 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 5581 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 5641 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 5701 tatggaaaaa cgccagcaac gcatcgataa aataaaagat tttatttagt ctccagaaaa 5761 aggggggaat gaaagacccc acctgtaggt ttggcaagct agcttaagta acgccatttt 5821 gcaaggcatg gaaaaataca taactgagaa tagagaagtt cagatcaagg tcaggaacag 5881 atggaacagc tgaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct 5941 cagggccaag aacagatgga acagctgaat atgggccaaa caggatatct gtggtaagca 6001 gttcctgccc cggctcaggg ccaagaacag atggtcccca gatgcggtcc agccctcagc 6061 agtttctaga gaaccatcag atgtttccag ggtgccccaa ggacctgaaa tgaccctgtg 6121 ccttatttga actaaccaat cagttcgctt ctcgcttctg ttcgcgcgct tctgctcccc 6181 gagctcaata aaagagccca caacccctca ctcggggcgc cagtcctccg attgactgag 6241 tcgcccgggt acccgtgtat ccaataaacc ctcttgcagt tgcatccgac ttgtggtctc 6301 gctgttcctt gggagggtct cctctgagtg attgactacc cgtcagcggg ggtctttcac 6361 atgcagcatg tatcaaaatt aatttggttt tttttcttaa gtatttacat taaatggcca 6421 tagtgcggcc gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 6481 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 6541 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 6601 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 6661 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 6721 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 6781 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 6841 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 6901 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 6961 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 7021 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 7081 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 7141 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 7201 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 7261 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 7321 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 7381 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 7441 acatttcccc gaaaagtgcc acct //
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