质粒名称: | pBV-Luc wt MBS1-4 |
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出品公司: | 爱迪基因 |
目录编号: | 16564 |
存储实验室: | Bert Vogelstein |
载体骨架基本信息 |
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载体名称: | pBV-Luc |
载体出品公司: | Vogelstein Lab |
载体抗性: | 氨苄青霉素 |
空载体大小: | 4867 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳动物表达,荧光素酶 |
筛选标记: | -- |
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: | c-MYC binding sites |
物种来源: | H. sapiens (human) |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | Luciferase(主干上的C末端) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | KpnI(未破坏) |
3' 克隆位点: | EcoRI (未破坏) |
5' 测序引物 | RVprimer3 |
3' 测序引物 | LucNrev |
文献引用方法 |
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材料和方法部分: | pBV-Luc wt MBS1-4 was a gift from Bert Vogelstein (Addgene plasmid # 16564 ; http://n2t.net/addgene:16564 ; RRID:Addgene_16564) |
参考文献部分: | Identification of CDK4 as a target of c-MYC. Hermeking H, Rago C, Schuhmacher M, Li Q, Barrett JF, Obaya AJ, O'Connell BC, Mateyak MK, Tam W, Kohlhuber F, Dang CV, Sedivy JM, Eick D, Vogelstein B, Kinzler KW. Proc Natl Acad Sci U S A. 2000 Feb 29. 97(5):2229-34. 10.1073/pnas.050586197 PubMed 10688915 |
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载体基本信息
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载体序列
LOCUS pBV-Luc wt MBS1-4 5025 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pBV-Luc wt MBS1-4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5025) AUTHORS Hermeking H, Rago C, Schuhmacher M, Li Q, Barrett JF, Obaya AJ, O'Connell BC, Mateyak MK, Tam W, Kohlhuber F, Dang CV, Sedivy JM, Eick D, Vogelstein B, Kinzler KW TITLE Identification of CDK4 as a target of c-MYC. JOURNAL Proc Natl Acad Sci U S A. 2000 Feb 29. 97(5):2229-34. PUBMED 10688915 REFERENCE 2 (bases 1 to 5025) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5025 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 289..298 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 295..1947 /codon_start=1 /gene="luc+" /product="firefly luciferase" /label=luciferase /note="enhanced luc+ version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" primer_bind complement(361..379) /label=LucNrev /note="Firefly luciferase, reverse primer" polyA_signal 1988..2109 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1999..2018) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 2053..2072 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(2357..2374) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2528..3116) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2608..2627) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3287..4147) /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 3910..3929 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4148..4252) /gene="bla" /label=AmpR promoter rep_origin 4279..4734 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4366..4385) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4576..4597 /label=F1ori-F /note="F1 origin, forward primer" polyA_signal 4865..4913 /note="synthetic polyadenylation signal" misc_feature 4927..5018 /label=pause site /note="RNA polymerase II transcriptional pause signal from the human alpha-2 globin gene" primer_bind 4967..4986 /label=RVprimer3 /note="pGL3 vector, forward primer" ORIGIN 1 ggtaccgggt tgttggcagc cagtcacgtg cccgccgcgt agccacacct ctgctcctca 61 gagcaatgtc aagcggtcac gtgtgatagc aacagatcac gtggctgcca tcgcccctcc 121 gcccccttac actcttcgcc ctcctcccag tcgaagcacc tcctgtccgc ccctcagcgc 181 catgggtggc ggtcacgtgc ccagaacgtc cggaattcgg gtatataatg gatccggtat 241 cgagatctgc gatctaagta agcttggcat tccggtactg ttggtaaagc caccatggaa 301 gacgccaaaa acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct 361 ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt 421 acagatgcac atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg 481 ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt 541 gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt 601 gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag 661 cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag 721 ctcccaatca tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag 781 tcgatgtaca cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg 841 ccagagtcct tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact 901 ggtctgccta aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc 961 agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca 1021 ttccatcacg gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc 1081 gtcttaatgt atagatttga agaagagctg tttctgagga gccttcagga ttacaagatt 1141 caaagtgcgc tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac 1201 aaatacgatt tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa 1261 gtcggggaag cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc 1321 actgagacta catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc 1381 ggtaaagttg ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg 1441 ggcgttaatc aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta 1501 aacaatccgg aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac 1561 atagcttact gggacgaaga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt 1621 aagtacaaag gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc 1681 aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg acgccggtga acttcccgcc 1741 gccgttgttg ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc 1801 gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta 1861 ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc 1921 aagaagggcg gaaagatcgc cgtgtaattc tagagtcggg gcggccggcc gcttcgagca 1981 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa 2041 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat 2101 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg 2161 gaggtttttt aaagcaagta aaacctctac aaatgtggta aaatcgataa ggatccgtcg 2221 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac 2281 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc 2341 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 2401 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 2461 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 2521 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 2581 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 2641 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 2701 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 2761 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 2821 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 2881 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 2941 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 3001 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 3061 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 3121 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 3181 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 3241 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 3301 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 3361 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 3421 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 3481 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 3541 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 3601 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 3661 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 3721 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 3781 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 3841 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 3901 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 3961 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 4021 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 4081 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 4141 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 4201 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 4261 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 4321 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 4381 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 4441 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg 4501 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 4561 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 4621 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 4681 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt 4741 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac 4801 gccagcccaa gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg 4861 ccgcaataaa atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat 4921 agtactaaca tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc 4981 tgtccccagt gcaagtgcag gtgccagaac atttctctat cgata //
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