pBluescript II KS(+)

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pBluescript II KS(+)载体 pBluescript II KS(+)质粒基本信息

载体名称: pBluescript II KS(+)
质粒类型: 大肠系列质粒;大肠克隆质粒;噬菌展示质粒
复制子: pBR322
载体大小: 2961bp
5' 测序引物及序列: T7(TAATACGACTCACTATAGGG)
3' 测序引物及序列: T3(ATTAACCCTCACTAAAGGGA)
载体标签: LacZ
载体抗性: 氨苄青霉素
克隆菌株: DH5α 等
备注: 蓝白斑筛选,产生单链DNA

pBluescript II KS(+)质粒图谱 pBluescript II KS(+)载体图谱和pBluescript II KS(+)载体序列 pBluescript II KS(+)质粒序列多克隆位点信息

pBluescript II KS(+)1载体图谱
pBluescript II KS(+)1载体图谱

pBluescript II KS(+)质粒 pBluescript II KS(+)载体简介

pBluescript II-KS(+)是大肠杆菌原核质粒。为克隆载体,其来源于噬菌体。该载体常被用在克隆和测序过程中,包括DNA测序中,嵌套缺失的构建;体外RNA转录,定点突变和基因定位。pBluescript II-KS(+)带有21个独特限制性内切酶的识别位点。在MCS区两侧是T7和T3 RNA聚合酶启动子序列,其主要帮助RNA的体外合成。pBluescript II SK(+)和 pBluescript II KS(+) 差异仅仅存在于多克隆位点的方向不同。 Standard cloning vector (phagemid excised from lambda ZAPII). The f1 (+) orientation allows rescue of sense strand ssDNA. pBluescript II KS(+) and pBluescript II SK(+) differ by the orientation of the MCS.

pBluescript II KS(+)质粒序列 pBluescript II KS(+)载体序列

LOCUS       pBluescript II KS(+)                2961 bp ds-DNA     circular SYN 13-JUN-2018
DEFINITION  synthetic circular DNA
KEYWORDS    pBluescript II KS (+)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2961)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..2961
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(241..816)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLISNSCSPGDPLV
                     LERPPPRWSSNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR
                     TDRPSQQLRSLNGEWDAPCSGALSAAGVVVTRSVTATLASALAPAPFAFFPSFLATFAG
                     FPRQALNRGLPLGFRFSALRHLDPKKLD"
     primer_bind     600..616
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    653..760
                     /note="MCS"
                     /note="pBluescript multiple cloning site"
     promoter        complement(773..791)
                     /note="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(812..828)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    836..852
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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)."
     promoter        complement(860..890)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    905..926
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1214..1802)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1973..2833)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(2834..2938)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa
      721 gcttatcgat accgtcgacc tcgagggggg gcccggtacc cagcttttgt tccctttagt
      781 gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      841 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      901 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      961 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     1021 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     1081 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     1141 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1201 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1261 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1321 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1381 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1441 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1501 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1561 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1621 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     1681 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1741 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1801 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1861 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1921 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1981 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2041 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2101 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2161 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2221 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2281 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2341 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2401 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2461 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2521 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2581 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2641 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2701 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2761 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2821 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2881 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2941 catttccccg aaaagtgcca c
//

      

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pBluescript II KS(+)质粒 pBluescript II KS(+)载体应用举例

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