质粒名称: | pCMV-Bx |
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出品公司: | 爱迪基因 |
目录编号: | 51552 |
存储实验室: | Michele Calos |
载体骨架基本信息 |
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载体名称: | pCMVSPORTbetaGal |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | -- |
高拷贝/低拷贝: | -- |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Bxb1 integrase |
物种来源: | -- |
插入大小: | -- |
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: | pCMV-Bx was a gift from Michele Calos (Addgene plasmid # 51552 ; http://n2t.net/addgene:51552 ; RRID:Addgene_51552) |
参考文献部分: | DICE, an efficient system for iterative genomic editing in human pluripotent stem cells. Zhu F, Gamboa M, Farruggio AP, Hippenmeyer S, Tasic B, Schule B, Chen-Tsai Y, Calos MP. Nucleic Acids Res. 2013 Dec 4. 10.1093/nar/gkt1290 PubMed 24304893 |
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载体基本信息
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载体简介
载体序列
LOCUS pCMV-Bx 5803 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses Bxb1 integrase in mammalian cells.. ACCESSION . VERSION . KEYWORDS pCMV-Bx SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5803) AUTHORS Zhu F, Gamboa M, Farruggio AP, Hippenmeyer S, Tasic B, Schule B, Chen-Tsai Y, Calos MP TITLE DICE, an efficient system for iterative genomic editing in human pluripotent stem cells. JOURNAL Nucleic Acids Res. 2013 Dec 4. PUBMED 24304893 REFERENCE 2 (bases 1 to 5803) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5803 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 2..457 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(89..108) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 299..320 /label=F1ori-F /note="F1 origin, forward primer" CDS 745..1605 /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(963..982) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" protein_bind complement(1707..1740) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." rep_origin 1818..2406 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2307..2326 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 2560..2577 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2907..2923 /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 2912..2934 /label=M13/pUC Reverse /note="In lacZ gene" enhancer 2953..3256 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 3257..3460 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 3411..3431 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 3457..3481 /label=LNCX /note="Human CMV promoter, forward primer" promoter 3576..3594 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 3576..3593 /label=SP6 /note="SP6 promoter, forward primer" polyA_signal 5546..5680 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5583..5602) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5637..5656 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc 61 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc 121 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt 181 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg 241 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt 301 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta 361 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt 421 aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt tcggggaaat 481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg 541 ccaggtcttg gactggtgag aacggcttgc tcggcagctt cgatgtgtgc tggagggaga 601 ataaaggtct aagatgtgcg atagagggaa gtcgcattga attatgtgct gtgtagggat 661 cgctggtatc aaatatgtgt gcccacccct ggcatgagac aataaccctg ataaatgctt 721 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc 781 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa 841 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt 901 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt 961 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc 1021 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg 1081 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg 1141 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac 1201 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca 1261 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta 1321 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat 1381 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa 1441 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag 1501 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat 1561 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt 1621 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg 1681 aagatccttt ttgataatct catgccataa cttcgtataa tgtatgctat acgaagttat 1741 ggcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 1801 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 1861 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 1921 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 1981 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 2041 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 2101 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 2161 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagca ttgagaaagc 2221 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 2281 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 2341 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 2401 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 2461 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag 2521 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa 2581 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc 2641 agagcttgca attcgcgcgt ttttcaatat tattgaagca tttatcaggg ttattgtctc 2701 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca 2761 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat 2821 aaaaataggc gtatacgagg ccctttcact cattaggcac cccaggcttt acactttata 2881 gcttccggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag 2941 catcgtgcag gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 3001 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 3061 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 3121 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 3181 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 3241 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 3301 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 3361 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 3421 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat 3481 cgcctggagg cgccatccac gctgttttga cctccataga agacaccggg accgatccag 3541 cctccggact ctagcctagg cttttgcaaa aagctattta ggtgacacta tagaaggtac 3601 gcctgcaggt accggtccgg aattcgccct tatgagagcc ctggtagtca tccgcctgtc 3661 ccgcgtcacc gatgctacga cttcaccgga gcgtcagctg gagtcttgcc agcagctctg 3721 cgcccagcgc ggctgggacg tcgtcggggt agcggaggat ctggacgtct ccggggcggt 3781 cgatccgttc gaccggaagc gcagaccgaa cctggcccgg tggctagcgt tcgaggagca 3841 accgttcgac gtgatcgtgg cgtaccgggt agaccggttg acccgatcga tccggcatct 3901 gcagcagctg gtccactggg ccgaggacca caagaagctg gtcgtctccg cgaccgaagc 3961 gcacttcgat acgacgacgc cgtttgcggc ggtcgtcatc gcgcttatgg gaacggtggc 4021 gcagatggaa ttagaagcga tcaaagagcg gaaccgttcg gctgcgcatt tcaatatccg 4081 cgccgggaaa taccgaggat ccctgccgcc gtggggatac ctgcctacgc gcgtggacgg 4141 ggagtggcgg ctggtgccgg accctgtgca gcgagagcgc atcctcgagg tgtatcaccg 4201 cgtcgtcgac aaccacgagc cgctgcacct ggtggcccac gacctgaacc ggcgtggtgt 4261 cctgtcgccg aaggactact tcgcgcagct gcaaggccgc gagccgcagg gccgggagtg 4321 gtcggctacc gcgctgaagc gatcgatgat ctccgaggcg atgctcgggt acgcgactct 4381 gaacggtaag accgtccgag acgacgacgg agccccgctg gtgcgggctg agccgatcct 4441 gacccgtgag cagctggagg cgctgcgcgc cgagctcgtg aagacctccc gggcgaagcc 4501 cgcggtgtct accccgtcgc tgctgctgcg ggtgttgttc tgcgcggtgt gcggggagcc 4561 cgcgtacaag ttcgccgggg gaggacgtaa gcacccgcgc taccgctgcc gctcgatggg 4621 gttcccgaag cactgcggga acggcacggt ggcgatggcc gagtgggacg cgttctgcga 4681 ggagcaggtg ctggatctgc tcggggacgc ggagcgtctg gagaaagtct gggtagccgg 4741 ctcggactcc gcggtcgaac tcgcggaggt gaacgcggag ctggtggacc tgacgtcgct 4801 gatcggctcc ccggcctacc gggccggctc tccgcagcga gaagcactgg atgcccgtat 4861 tgcggcgctg gccgcgcggc aagaggagct ggagggtcta gaggctcgcc cgtctggctg 4921 ggagtggcgc gagaccgggc agcggttcgg ggactggtgg cgggagcagg acaccgcggc 4981 aaagaacacc tggcttcggt cgatgaacgt tcggctgacg ttcgacgtcc gcggcgggct 5041 gactcgcacg atcgacttcg gggatctgca ggagtacgag cagcatctca ggctcggcag 5101 cgtggtcgaa cggctacaca ccgggatgtc gtagaagggc gaattcccgg gtcgacgagc 5161 tcactagctt gggatctttg tgaaggaacc ttacttctgt ggtgtgacat aattggacaa 5221 actacctaca gagatttaaa gctctaaggt aaatataaaa tttttaagtg tataatgtgt 5281 taaactagct gcatatgctt gctgcttgag agttttgctt actgagtatg atttatgaaa 5341 atattataca caggagctag tgattctaat tgtttgtgta ttttagattc acagtcccaa 5401 ggctcatttc aggcccctca gtcctcacag tctgttcatg atcataatca gccataccac 5461 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca 5521 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata 5581 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg 5641 tttgtccaaa ctcatcaatg tatcttatca tgtctggatc gatcctgcat taatgaatcg 5701 gccaacgcgc ggggagaggc ggtttgcgta ttggctggcg taatagcgaa gaggcccgca 5761 ccgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atg //