载体名称: | pBluescript II KS(+) |
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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 |
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pBluescript II KS(+)载体 pBluescript II KS(+)质粒基本信息
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(+)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 //