基本信息

复制子	p15A
质粒分类	大肠系列质粒；大肠克隆质粒；基因克隆质粒
质粒大小	4245bp
原核抗性	Chl/Tet
克隆菌株	DH5α
培养条件	37℃
5'测序引物	Tcr-F（GAAGTCATGCGCCGGTTAAG）
3'测序引物	pBRrevBam-R (GGTGATGTCGGCGATATAGG)
备注	丰富的酶切位点
质粒宿主	大肠杆菌
质粒用途	基因模板
片段类型	基因CDS
片段物种	空载体

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质粒简介

        pACYC184质粒是一种原核表达载体，含有p15A复制起点的大肠杆菌质粒克隆载体。这允许pACYC184在与ColE1相容性组的质粒（例如pBR322，pUC19）的细胞中共存。它是一个低拷贝数载体，每个细胞约15个拷贝，但可以用壮观霉素放大。由于cat基因的存在，氯霉素不能用于扩增。

        pACYC184 is an E. coli plasmid cloning vector containing the p15A origin of replication (1-4). This allows pACYC184 to coexist in cells with plasmids of the ColE1 compatibility group (e.g., pBR322, pUC19). It is a low copy number vector, at about 15 copies per cell (5), but can be amplifed with spectinomycin. Chloramphenicol cannot be used for amplifcation due to the presence of the cat gene.

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质粒图谱

质粒序列

LOCUS       Exported                4245 bp ds-DNA     circular SYN 26-8-2015
DEFINITION  synthetic circular DNA
KEYWORDS    Untitled 2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4245)
  TITLE     Direct Submission
  JOURNAL   Exported 2015-8-26 from MiaoLingPlasmid
            http://www.shhebio.com
FEATURES             Location/Qualifiers
     source          1..4245
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(220..322)
                     /note="cat promoter"
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /note="p15A ori"
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells
                     that contain a second plasmid with the ColE1 origin."
     promoter        1505..1533
                     /note="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             1581..2771
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /note="TcR"
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     CDS             complement(3805..219)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagcttt aatgcggtag tttatcacag ttaaattgct
     1561 aacgcagtca ggcaccgtgt atgaaatcta acaatgcgct catcgtcatc ctcggcaccg
     1621 tcaccctgga tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg
     1681 atatcgtcca ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt
     1741 tgatgcaatt tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc
     1801 cagtcctgct cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac
     1861 ccgtcctgtg gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg
     1921 cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg
     1981 ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt
     2041 tgggcgccat ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc
     2101 tactactggg ctgcttccta atgcaggagt cgcataaggg agagcgtcga ccgatgccct
     2161 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     2221 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     2281 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     2341 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     2401 tcggcgagaa gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc
     2461 tggcgttcgc gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg
     2521 gcatcgggat gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg
     2581 gacagcttca aggatcgctc gcggctctta ccagcctaac ttcgatcact ggaccgctga
     2641 tcgtcacggc gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag
     2701 gcgccgccct ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca
     2761 cctcgacctg aatggaagcc ggcggcacct cgctaacgga ttcaccactc caagaattgg
     2821 agccaatcaa ttcttgcgga gaactgtgaa tgcgcaaacc aacccttggc agaacatatc
     2881 catcgcgtcc gccatctcca gcagccgcac gcggcgcatc tcgggcagcg ttgggtcctg
     2941 gccacgggtg cgcatgatcg tgctcctgtc gttgaggacc cggctaggct ggcggggttg
     3001 ccttactggt tagcagaatg aatcaccgat acgcgagcga acgtgaagcg actgctgctg
     3061 caaaacgtct gcgacctgag caacaacatg aatggtcttc ggtttccgtg tttcgtaaag
     3121 tctggaaacg cggaagtccc ctacgtgctg ctgaagttgc ccgcaacaga gagtggaacc
     3181 aaccggtgat accacgatac tatgactgag agtcaacgcc atgagcggcc tcatttctta
     3241 ttctgagtta caacagtccg caccgctgtc cggtagctcc ttccggtggg cgcggggcat
     3301 gactatcgtc gccgcactta tgactgtctt ctttatcatg caactcgtag gacaggtgcc
     3361 ggcagcgccc aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc
     3421 gccctgcacc attatgttcc ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     3481 acctacatct gtattaacga agcgctaacc gtttttatca ggctctggga ggcagaataa
     3541 atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg gcctcaggca
     3601 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     3661 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg
     3721 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag gcgtttaagg
     3781 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg
     3841 taattcatta agcattctgc cgacatggaa gccatcacag acggcatgat gaacctgaat
     3901 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     3961 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     4021 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     4081 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     4141 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     4201 aacactatcc catatcacca gctcaccgtc tttcattgcc atacg
//