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pKK223-3

pKK223-3
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  • 产品名称:pKK223-3
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简单介绍
pKK223-3菌株发货前均经过严格的多重验证,如存在质量问题,请在收到产品的三个月内通知我司。pKK223-3收到质粒后请短暂离心,加入20μl ddH2O溶解质粒,取2μl转化至对应感受态中,挑取单克隆重新提取质粒后使用��
产品描述

pKK223-3

  • 产品信息
  • 基本信息
  • 质粒简介
  • 质粒图谱
  • 质粒序列

产品信息

产品货号 产品名称 产品规格 优惠价
XY0084 pKK223-3

20μl

¥请询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证,如存在质量问题,请在收到产品的三个月内通知我司。收到质粒后请短暂离心,加入20μl ddH2O溶解质粒,取2μl转化至对应感受态中,挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

启动子: Tac
复制子: pBR322 ori
终止子: rrnB T1
质粒分类: 大肠杆菌载体;蛋白表达质粒
质粒大小: 4584bp
原核抗性: 氨苄青霉素Amp
克隆菌株: DH5α
培养条件: 37℃,有氧,LB
表达宿主: 大肠杆菌
培养条件: 37℃,有氧,LB
诱导方式: IPTG或乳糖及其类似物
备注: 表达质粒

质粒简介

pKK223-3 (1) contains the strong tac promoter upstream from the multiple cloning site and the strong rrnB ribosomal terminator downstream for control of protein expression (2-4). Induction: tac promoter is inducible with 1-5 mM IPTG. Promoter is very strong; even uninduced cells may show a low level of expression. Expression: Genes containing a ribosome-binding site and ATG can be inserted into any unique site in the MCS for expression. The ribosomebinding site on the plasmid can be utilised to express inserts if the ATG start codon of the insert is within 5-13 bp from the provided ribosomebindingsite (5, 6). Transcription terminators: rrnB transcription terminators stabilize the plasmid by inhibiting read-through transcription initiated from the tac promoter in the parent plasmid. Host(s): lac Iq strains; E. coli JM105 is recommended. Selectable marker(s): Plasmid confers resistance to 100 μg/ml ampicillin. Amplification: Recommended. Control Regions: Expression control region: tac promoter: -10: 50-44; -35: 72-67; Ribosome binding site: 11. MCS: 4552-4587. rrnB operon region: 5S rRNA region: 4471-4352; rrnB T1 terminator: 4348-4305; rrnB T2 terminator: 4173-4146. b-lactamase gene region: Promoter: -10: 4082-4077; -35: 4105-4100; Start codon (ATG): 4035; Stop codon (TAA): 3178. Plasmid replication region: Site of replication initiation: 2417-2415; Region necessary for replication: 3111-2415.

 

质粒图谱

质粒序列

LOCUS       Exported                4584 bp ds-DNA     circular SYN 18-九月-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4584)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2016年9月18日 from SnapGene Viewer 2.8.1
            http://www.miaolingbio.com
FEATURES             Location/Qualifiers
     source          1..4584
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    20..36
                     /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)."
     misc_feature    complement(44..72)
                     /note="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     CDS             1796..1987
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    2089..2229
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2415..3003)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     promoter        complement(4034..4125)
                     /gene="bla"
                     /note="AmpR promoter"
     terminator      4144..4171
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      4263..4349
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacattata cgagccgatg
       61 attaattgtc aacagctcat ttcagaatat ttgccagaac cgttatgatg tcggcgcaaa
      121 aaacattatc cagaacggga gtgcgccttg agcgacacga attatgcagt gatttacgac
      181 ctgcacagcc ataccacagc ttccgatggc tgcctgacgc cagaagcatt ggtgcaccgt
      241 gcagtcgata agcccggatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca
      301 caggtgcggt tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc
      361 acttcgggct catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg
      421 gactgttggg cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc
      481 tcaacctact actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgaccga
      541 tgcccttgag agccttcaac ccagtcagct ccttccggtg ggcgcggggc atgactatcg
      601 tcgccgcact tatgactgtc ttctttatca tgcaactcgt aggacaggtg ccggcagcgc
      661 tctgggtcat tttcggcgag gaccgctttc gctggagcgc gacgatgatc ggcctgtcgc
      721 ttgcggtatt cggaatcttg cacgccctcg ctcaagcctt cgtcactggt cccgccacca
      781 aacgtttcgg cgagaagcag gccattatcg ccggcatggc ggccgacgcg ctgggctacg
      841 tcttgctggc gttcgcgacg cgaggctgga tggccttccc cattatgatt cttctcgctt
      901 ccggcggcat cgggatgccc gcgttgcagg ccatgctgtc caggcaggta gatgacgacc
      961 atcagggaca gcttcaagga tcgctcgcgg ctcttaccag cctaacttcg atcactggac
     1021 cgctgatcgt cacggcgatt tatgccgcct cggcgagcac atggaacggg ttggcatgga
     1081 ttgtaggcgc cgccctatac cttgtctgcc tccccgcgtt gcgtcgcggt gcatggagcc
     1141 gggccacctc gacctgaatg gaagccggcg gcacctcgct aacggattca ccactccaag
     1201 aattggagcc aatcaattct tgcggagaac tgtgaatgcg caaaccaacc cttggcagaa
     1261 catatccatc gcgtccgcca tctccagcag ccgcacgcgg cgcatctcgg gcagcgttgg
     1321 gtcctggcca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
     1381 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
     1441 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
     1501 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
     1561 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
     1621 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
     1681 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
     1741 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
     1801 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
     1861 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
     1921 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
     1981 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     2041 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     2101 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     2161 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2221 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2281 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2341 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2401 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2461 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2521 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2581 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg
     2641 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2701 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2761 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2821 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     2881 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2941 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     3001 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     3061 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     3121 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     3181 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     3241 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     3301 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     3361 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     3421 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     3481 gccattgcta cagcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     3541 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     3601 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3661 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3721 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3781 cggcgtcaac acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3841 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3901 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3961 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     4021 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     4081 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaaagagt ttgtagaaac
     4141 gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc agtttatggc
     4201 gggcgtcctg cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc cgctcccggc
     4261 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     4321 tctttcgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg
     4381 agaccccaca ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg
     4441 tgggaccacc gcgctactgc cgccaggcaa attctgtttt atcagaccgc ttctgcgttc
     4501 tgatttaatc tgtatcaggc tgaaaatctt ctctcatccg ccaaaacagc caagcttggc
     4561 tgcaggtcga cggatccccg ggaa
//
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