pUC19大肠克隆质粒

基本信息

启动子:	Lac/lac promoter
复制子:	ColE1 ori
质粒分类:	大肠杆菌载体；克隆质粒
质粒大小:	2686bp
质粒标签:	LacZ
原核抗性:	氨苄青霉素Amp
克隆菌株:	DH5α
培养条件:	37℃，有氧，LB
5'测序引物:	M13F:TGTAAAACGACGGCCAGT
3'测序引物:	M13R:CAGGAAACAGCTATGACC
备注:	蓝白斑筛选

质粒简介

　     pUC18 和 pUC19 大小只有 2686bp ，是最常用的质粒载体，其结构组成紧凑，几乎不含多余的 DNA片段，GenBank注册号为 L08752(pUC18)和 X02514(pUC19)。由 pBR322 改造而来，其中 lacZ (MSC)来自 M13mp18/19  。这两个质粒的结构几乎是完全一样的，只是多克隆位点的排列方向相反。这些质粒缺乏控制拷贝数的 rop 基因，因此其拷贝数达 500-700 。 pUC 系列载体含有一段 lacZ 蛋白氨基末端的部分编码序列，在特定的受体细胞中可表现 α-互补作用。因此在多克隆位点中插入了外源片段后，可通过 α-互补作用形成的蓝色和白色菌落筛选重组 质粒。pUC18、pUC19载体适合于DNA片段的克隆、进行DNA测序、对外源基因进行表达等。由于在lacZ领域中含有多克隆位点，因此在以lacZ缺欠细胞作为宿主 (如：JM109等) 进行转化后，在含有IPTG、X-Gal 的平板培养基上进行培养时，可以很容易地通过蓝白筛选，判断载体中有无DNA片段的插入。同时还可以通过载体上的lac promoter表达外源基因，对插入载体中的DNA片段进行测序等。进行DNA测序时，可以方便地使用M13系列的通用引物。

质粒图谱

质粒序列

LOCUS       Exported File           2686 bp ds-DNA    circular SYN 02-2-2015
KEYWORDS    pUC19
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2686)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2015-2-2 from MLCC
FEATURES             Location/Qualifiers
     source          1..2686
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    106..127
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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)."
     primer_bind     204..220
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             216..539
                     /codon_start=1
                     /gene="lacZ?fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    233..289
                     /gene="
                     "
                     /note="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(290..306)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        780..884
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             885..1745
                     /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"
     rep_origin      1916..2504
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttgc
      241 atgcctgcag gtcgactcta gaggatcccc gggtaccgag ctcgaattca ctggccgtcg
      301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      421 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
      481 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
      541 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
      601 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
      661 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
      721 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
      781 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
      841 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
      901 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
      961 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1021 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1081 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1141 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1201 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1261 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1321 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     1381 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     1441 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     1501 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     1561 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     1621 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     1681 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     1741 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     1801 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     1861 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1921 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1981 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2041 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2101 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2161 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2221 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2281 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2521 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2581 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2641 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaaga