pET-30 EK/LIC

价格:1000元

联系方式:I47-825O-882O

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pET30 Xa/LIC载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET30 EK/LIC, pet 30a EK/LIC
质粒类型: 大肠杆菌蛋白表达
表达水平:
克隆方法: 多克隆位点,限制性内切酶
载体大小: 5439 bp
5' 测序引物: T7
5' 测序引物序列:   5'-TAATACGACTCACTATAGGG-3'
载体标签: N-His, N-S, N-EK, C-His
载体抗性: Kanamycin
表达宿主菌: BL21(DE3) Rosetta2(DE3)C43(DE3)
备注: For directional cloning of PCR-amplified DNA; ligation independent cloning; Factor Xa cleavage site
产品目录号: 69077-3
稳定性: 瞬时表达 Transient
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒

pET30 Xa/LIC载体质粒图谱和多克隆位点信息

pet30 EK/LIC载体图谱
pet30 EK/LIC多克隆位点

pet30 EK/LIC载体图谱

Feature Name Start End  
T7_terminator 129 1
T7_Terminal_primer 69 87
EK 250 236
lacI 319 1925
T7_transl_en_RBS 389 373
lacO 434 407
T7_promoter 452 434
tet (300 - 563) 488 751
pBRrevBam_primer 559 540
tet (576 - 851) 1984 2259
ROP 2734 2925
pGEX_3_primer 2941 2919
pBR322_origin 3959 3340
KanR2 4065 4880
ORF Start End  
ORF frame 2 344 1093
ORF frame 3 966 1925
ORF frame 3 4065 4880
Enzyme Name Cut
XhoI 158
NotI 166
EagI 166
HindIII 173
SalI 179
SacI 190
EcoRI 192
BamHI 198
EcoRV 206
NcoI 211
KpnI 255
BglII 258
BstBI 285
NdeI 363
XbaI 401
BclI 1207
ApaI 1404
HpaI 1699
FspI 2275
NruI 4153

pET30 Xa/LIC载体简介

The pET-30 Ek/LIC vector is prepared for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides. Using specifically designed primers for amplification and the pET-30 Ek/LIC Cloning Kit (Cat. No. 69077-3), inserts can be efficiently cloned without the need for restriction digestion or ligation. Fusion proteins contain N-terminal cleavable His•Tag® and S•Tag™ sequences for detection and purification. Unique sites are shown on the circle map. Note that the sequence is numbered by the pBR322 convention, so the T7 expression region is reversed on the circle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymerase is shown below. The f1 origin is oriented so that infection with helper phage will produce virions containing single stranded DNA that corresponds to the coding strand. Therefore, single stranded sequencing should be performed using the T7 terminator primer (Cat. No. 69337-3).

pET30 Xa/LIC载体序列

LOCUS       pET-30 Ek/LIC    5439 bp     DNA Circular    SYN
DEFINITION  pET-30 Ek/LIC
ACCESSION   
KEYWORDS    
SOURCE      
  ORGANISM  other sequences; artificial sequences; vectors.
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|biofeng.com|
COMMENT     VNTOAUTHORAD1|Novagen Inc.|
FEATURES             Location/Qualifiers
     source          1..5439
                     /organism="pET-30 Ek/LIC"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    complement(236..250)
                     /label="EK"
     misc_feature    319..1925
                     /label="lacI"
     CDS             344..1093
                     /label="ORF frame 2"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(373..389)
                     /label="T7_transl_en_RBS"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(407..434)
                     /label="lacO"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     promoter        complement(434..452)
                     /label="T7_promoter"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     gene            488..751
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(540..559)
                     /label="pBRrevBam_primer"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     CDS             966..1925
                     /label="ORF frame 3"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            1984..2259
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    2734..2925
                     /label="ROP"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2919..2941)
                     /label="pGEX_3_primer"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      complement(3340..3959)
                     /label="pBR322_origin"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            4065..4880
                     /label="KanR2"
                     /gene="KanR2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             4065..4880
                     /label="ORF frame 3"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLY
                     GKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIP
                     GKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGL
                     VDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVG
                     RVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"
ORIGIN
    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
  121 tgttagcagc cggatctcag tggtggtggt ggtggtgctc gagtgcggcc gcaagcttgt
  181 cgacggagct cgaattcgga tccgatatcg ccatggttga ggagaagccc gggctcttgt
  241 cgtcgtcatc ggtacccaga tctgggctgt ccatgtgctg gcgttcgaat ttagcagcag
  301 cggtttcttt cataccagaa ccgcgtggca ccagaccaga agaatgatga tgatgatggt
  361 gcatatgtat atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc
  421 cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat cgatctcgat
  481 cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc
  541 ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc
  601 ttgtttcggc gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc
  661 cttgcatgca ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg
  721 cttcctaatg caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc
  781 gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga
  841 atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg
  901 tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag
  961 cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac
 1021 agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg
 1081 tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag
 1141 aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca
 1201 gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct
 1261 gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta
 1321 ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc
 1381 agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg
 1441 gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact
 1501 ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca
 1561 ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt
 1621 ccgggctgcg cgttggtgcg gacatctcgg tagtgggata cgacgatacc gaagacagct
 1681 catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca
 1741 gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc
 1801 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
 1861 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
 1921 agtgagcgca acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg
 1981 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
 2041 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
 2101 ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg
 2161 cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc
 2221 aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg
 2281 atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag
 2341 aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc
 2401 tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag
 2461 tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt
 2521 ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg
 2581 tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt
 2641 tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc
 2701 atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt
 2761 aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg
 2821 gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac
 2881 cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
 2941 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
 3001 tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga
 3061 gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat
 3121 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc
 3181 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
 3241 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
 3301 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
 3361 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
 3421 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
 3481 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
 3541 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
 3601 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
 3661 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
 3721 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
 3781 cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
 3841 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
 3901 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
 3961 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa
 4021 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac
 4081 gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
 4141 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga
 4201 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
 4261 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
 4321 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag
 4381 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
 4441 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
 4501 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
 4561 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
 4621 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
 4681 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
 4741 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
 4801 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
 4861 tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag
 4921 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta
 4981 aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
 5041 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
 5101 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
 5161 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
 5221 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
 5281 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
 5341 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
 5401 gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca
//

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pET30 Xa/LIC载体应用举例

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