首页 » pET30 EK/LIC大肠杆菌蛋白表达载体-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

pET30 EK/LIC大肠杆菌蛋白表达载体-BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心

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  • 货  号:pET30 EK/LIC vector
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pET30 EK/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 Ek-LIC大肠杆菌表达载体,PCR产物直接连接表达载体,无需酶切酶连,BioVector NTCC质粒载体菌种细胞蛋白抗体基因保藏中心提供

pET30 EK/LIC载体质粒图谱和多克隆位点信息


Feature NameStartEnd
T7_terminator1291
T7_Terminal_primer6987
EK250236
lacI3191925
T7_transl_en_RBS389373
lacO434407
T7_promoter452434
tet (300 - 563)488751
pBRrevBam_primer559540
tet (576 - 851)19842259
ROP27342925
pGEX_3_primer29412919
pBR322_origin39593340
KanR240654880
ORFStartEnd
ORF frame 23441093
ORF frame 39661925
ORF frame 340654880
Enzyme NameCut
XhoI158
NotI166
EagI166
HindIII173
SalI179
SacI190
EcoRI192
BamHI198
EcoRV206
NcoI211
KpnI255
BglII258
BstBI285
NdeI363
XbaI401
BclI1207
ApaI1404
HpaI1699
FspI2275
NruI4153


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.biovector.net/
COMMENT     ORIGDB|GenBank
COMMENT     VNTAUTHORNAME|www.biovector.net|
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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