| 載體名稱: | pBluescript II KS(+) |
|---|---|
| 質粒類型: | 大腸系列質粒；大腸克隆質粒；噬菌展示質粒 |
| 復制子: | pBR322 |
| 載體大小: | 2961bp |
| 5' 測序引物及序列: | T7（TAATACGACTCACTATAGGG） |
| 3' 測序引物及序列: | T3（ATTAACCCTCACTAAAGGGA) |
| 載體標簽: | LacZ |
| 載體抗性: | 氨芐青霉素 |
| 克隆菌株: | DH5α 等 |
| 備注: | 藍白斑篩選，產生單鏈DNA |
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pBluescript II KS(+)載體 pBluescript II KS(+)質粒基本信息
pBluescript II KS(+)質粒圖譜 pBluescript II KS(+)載體圖譜和pBluescript II KS(+)載體序列 pBluescript II KS(+)質粒序列多克隆位點信息
1載體圖譜.png)
1載體圖譜.jpg)
pBluescript II KS(+)質粒 pBluescript II KS(+)載體簡介
pBluescript II-KS(+)是大腸桿菌原核質粒。為克隆載體，其來源于噬菌體。該載體常被用在克隆和測序過程中，包括DNA測序中，嵌套缺失的構建；體外RNA轉錄，定點突變和基因定位。pBluescript II-KS(+)帶有21個獨特限制性內切酶的識別位點。在MCS區兩側是T7和T3 RNA聚合酶啟動子序列，其主要幫助RNA的體外合成。pBluescript II SK(+)和 pBluescript II KS(+) 差異僅僅存在于多克隆位點的方向不同。 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(+)質粒序列 pBluescript II KS(+)載體序列
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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