pCMV-Bx

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联系方式:I47-825O-882O

相关技术服务:质粒构建    基因合成    质粒大提

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载体基本信息

质粒名称: pCMV-Bx 
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
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
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
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

载体图谱质粒图谱和多克隆位点信息

pCMV-Bx质粒图谱

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
//

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