| 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 |
买家导航
pkk223-3载体质粒基本信息
pkk223-3质粒图谱载体图谱和pkk223-3载体序列质粒序列多克隆位点信息
pkk223-3质粒载体简介
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.
pkk223-3质粒序列载体序列
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
//
