pkk223-3

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载体基本信息

出品公司: PL-Pharmacia
载体名称: pkk223-3
质粒类型: 大肠杆菌表达载体
高拷贝/低拷贝: --
启动子: --
克隆方法: 多克隆位点,限制性内切酶
载体大小: 4586 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: --
载体抗性: Ampicillin
筛选标记: --
备注: --
产品目录号: --
稳定性: --
组成型: --
病毒/非病毒: 非病毒

载体图谱质粒图谱和多克隆位点信息

pkk223-3载体图谱

载体简介

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.

载体序列

LOCUS       pKK223-3	4586 bp 	DNA	SYN
DEFINITION  pKK223-3
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
FEATURES             Location/Qualifiers
     source          1..4586
                     /organism="pKK223-3"
                     /mol_type="other DNA"
     misc_feature    complement(9..31)
                     /label="M13_pUC_rev_primer"
     promoter        complement(43..71)
                     /label="tac_promoter"
     gene            268..1158
                     /label="tet (300 - 1190)"
                     /gene="tet (300 - 1190)"
     misc_feature    complement(320..339)
                     /label="pBRrevBam_primer"
     CDS             373..1158
                     /label="ORF frame 1"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIMQLVGQVPAALWVIFG
                     EDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAIIAGMAADALGYVLLA
                     FATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSLAALTSLTSITGPL
                     IVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST*"
     CDS             complement(505..963)
                     /label="ORF frame 3"
                     /translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
                     RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
                     CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSTLSLMRLLH*"
     misc_feature    1799..1990
                     /label="ROP"
                     /translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
                     RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
                     CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSTLSLMRLLH*"
     misc_feature    complement(1984..2006)
                     /label="pGEX_3_primer"
                     /translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
                     RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
                     CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSTLSLMRLLH*"
     gene            complement(3177..4037)
                     /label="Ampicillin"
                     /gene="Ampicillin"
                     /translation="MVVIYLPGQHGLQRGHPDAAGSEKNHNGEGHPASRRERQQDVAQ
                     RVGRHAGDNGLLLAETFGGGTSDEGLSEGVQDSEYRKRQADHRRAPAKAVLAENDPER
                     CRHLSYELHDKEDSHKCGDDSHAPRPPEGADWVEGSQGHRSTLSLMRLLH*"
     CDS             complement(3177..4037)
                     /label="ORF frame 1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     promoter        complement(4079..4107)
                     /label="AmpR_promoter"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     terminator      complement(4148..4175)
                     /label="rrnB_T2_terminator"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     terminator      complement(4307..4350)
                     /label="rrnB_T1_terminator"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     terminator      complement(4316..4473)
                     /label="rrnB_terminator"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    4506..4523
                     /label="pBAD_rev_primer"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
     misc_feature    4506..4523
                     /label="pTrcHis_rev_primer"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW*"
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 agctccggat cctctacgcc ggacgcatcg tggccggcat caccggcgcc
  301 acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc
  361 cacttcgggc tcatgagcgc ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg
  421 ggactgttgg gcgccatctc cttgcatgca ccattccttg cggcggcggt gctcaacggc
  481 ctcaacctac tactgggctg cttcctaatg caggagtcgc ataagggaga gcgtcgaccg
  541 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
  601 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
  661 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
  721 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
  781 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc gctgggctac
  841 gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat tcttctcgct
  901 tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt agatgacgac
  961 catcagggac agcttcaagg atcgctcgcg gctcttacca gcctaacttc gatcactgga
 1021 ccgctgatcg tcacggcgat ttatgccgcc tcggcgagca catggaacgg gttggcatgg
 1081 attgtaggcg ccgccctata ccttgtctgc ctccccgcgt tgcgtcgcgg tgcatggagc
 1141 cgggccacct cgacctgaat ggaagccggc ggcacctcgc taacggattc accactccaa
 1201 gaattggagc caatcaattc ttgcggagaa ctgtgaatgc gcaaaccaac ccttggcaga
 1261 acatatccat cgcgtccgcc atctccagca gccgcacgcg gcgcatctcg ggcagcgttg
 1321 ggtcctggcc acgggtgcgc atgatcgtgc tcctgtcgtt gaggacccgg ctaggctggc
 1381 ggggttgcct tactggttag cagaatgaat caccgatacg cgagcgaacg tgaagcgact
 1441 gctgctgcaa aacgtctgcg acctgagcaa caacatgaat ggtcttcggt ttccgtgttt
 1501 cgtaaagtct ggaaacgcgg aagtcagcgc cctgcaccat tatgttccgg atctgcatcg
 1561 caggatgctg ctggctaccc tgtggaacac ctacatctgt attaacgaag cgctggcatt
 1621 gaccctgagt gatttttctc tggtcccgcc gcatccatac cgccagttgt ttaccctcac
 1681 aacgttccag taaccgggca tgttcatcat cagtaacccg tatcgtgagc atcctctctc
 1741 gtttcatcgg tatcattacc cccatgaaca gaaattcccc cttacacgga ggcatcaagt
 1801 gaccaaacag gaaaaaaccg cccttaacat ggcccgcttt atcagaagcc agacattaac
 1861 gcttctggag aaactcaacg agctggacgc ggatgaacag gcagacatct gtgaatcgct
 1921 tcacgaccac gctgatgagc tttaccgcag ctgcctcgcg cgtttcggtg atgacggtga
 1981 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
 2041 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
 2101 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
 2161 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
 2221 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
 2281 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
 2341 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
 2401 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
 2461 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
 2521 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
 2581 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
 2641 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
 2701 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
 2761 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
 2821 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
 2881 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
 2941 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
 3001 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
 3061 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
 3121 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
 3181 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
 3241 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
 3301 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
 3361 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
 3421 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
 3481 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
 3541 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
 3601 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
 3661 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
 3721 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
 3781 tgcccggcgt caacacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
 3841 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
 3901 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
 3961 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
 4021 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
 4081 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaaa gagtttgtag
 4141 aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc tggcagttta
 4201 tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc
 4261 cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc
 4321 ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct actctcgcat
 4381 ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc ggcatggggt
 4441 caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga ccgcttctgc
 4501 gttctgattt aatctgtatc aggctgaaaa tcttctctca tccgccaaaa cagaagcttg
 4561 gctgcaggtc gacggatccc cgggaa
//

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