pET-30c(+)

价格:800元

联系方式:I47-825O-882O

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pET30c载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET30c, pet 30c
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: 5423 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-Thrombin, N-EK, C-His
载体抗性: Kanamycin
表达宿主菌: BL21(DE3) Rosetta2(DE3)C43(DE3)
备注: N term thrombin cleavage site; N term enterokinase cleavage site; a,b,c vary by MCS
产品目录号: 69911-3
稳定性: 瞬时表达 Transient
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒

pET30c载体质粒图谱和多克隆位点信息

pet30c载体图谱
pet30c多克隆位点

Feature Name Start End  
T7_terminator 129 1
T7_Terminal_primer 69 87
EK 231 220
lacI 303 1909
T7_transl_en_RBS 373 357
lacO 418 391
T7_promoter 436 418
tet (300 - 563) 472 735
pBRrevBam_primer 543 524
tet (576 - 851) 1968 2243
ROP 2718 2909
pGEX_3_primer 2925 2903
pBR322_origin 3943 3324
KanR2 4049 4864
ORF Start End  
ORF frame 1 328 1077
ORF frame 2 950 1909
ORF frame 2 4049 4864
Enzyme Name Cut
XhoI 158
NotI 166
EagI 166
HindIII 173
SalI 179
SacI 190
EcoRI 192
BamHI 198
EcoRV 209
NcoI 213
KpnI 239
BglII 242
BstBI 269
NdeI 347
XbaI 385
BclI 1191
ApaI 1388
HpaI 1683
FspI 2259
NruI 4137
SmaI 4354
XmaI 4352

pET30c载体简介

The pET-30a-c(+) vectors carry an N-terminal His• Tag®/ thrombin/ S• Tag™/ enterokinase configuration plus an optional C-terminal His• Tag sequence. 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 circular 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).

pET30c载体序列

LOCUS       pET-30 c (+)    5423 bp     DNA Circular    SYN
DEFINITION  pET-30 c (+)
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..5423
                     /organism="pET-30 c (+)"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    complement(220..231)
                     /label="EK"
     misc_feature    303..1909
                     /label="lacI"
     CDS             328..1077
                     /label="ORF frame 1"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(357..373)
                     /label="T7_transl_en_RBS"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(391..418)
                     /label="lacO"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     promoter        complement(418..436)
                     /label="T7_promoter"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     gene            472..735
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(524..543)
                     /label="pBRrevBam_primer"
                     /translation="MMMMMVHMYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSI
                     SILYAGRIVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLL
                     GAISLHAPFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRE
                     SIQGGECETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPR
                     GTTTGGQTVVADWRCHLQSGPARAVANCRGD*"
     CDS             950..1909
                     /label="ORF frame 2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            1968..2243
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    2718..2909
                     /label="ROP"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2903..2925)
                     /label="pGEX_3_primer"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      complement(3324..3943)
                     /label="pBR322_origin"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            4049..4864
                     /label="KanR2"
                     /gene="KanR2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             4049..4864
                     /label="ORF frame 2"
                     /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 tccacagata tcccatggcc ttgtcgtcgt cgtcggtacc
  241 cagatctggg ctgtccatgt gctggcgttc gaatttagca gcagcggttt ctttcatacc
  301 agaaccgcgt ggcaccagac cagaagaatg atgatgatga tggtgcatat gtatatctcc
  361 ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca caattcccct
  421 atagtgagtc gtattaattt cgcgggatcg agatcgatct cgatcctcta cgccggacgc
  481 atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc
  541 accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt
  601 atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc
  661 cttgcggcgg cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag
  721 tcgcataagg gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg
  781 tatggcatga tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac
  841 gttatacgat gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa
  901 ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct
  961 gaattacatt cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg
 1021 cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc
 1081 tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga
 1141 agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa
 1201 ctatccgctg gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc
 1261 gttatttctt gatgtctctg accagacacc catcaacagt attattttct cccatgaaga
 1321 cggtacgcga ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt
 1381 agcgggccca ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct
 1441 cactcgcaat caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg
 1501 ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc
 1561 caacgatcag atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg
 1621 tgcggacatc tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc
 1681 gttaaccacc atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct
 1741 gcaactctct cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa
 1801 aagaaaaacc accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
 1861 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
 1921 ttaatgtaag ttagctcact cattaggcac cgggatctcg accgatgccc ttgagagcct
 1981 tcaacccagt cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga
 2041 ctgtcttctt tatcatgcaa ctcgtaggac aggtgccggc agcgctctgg gtcattttcg
 2101 gcgaggaccg ctttcgctgg agcgcgacga tgatcggcct gtcgcttgcg gtattcggaa
 2161 tcttgcacgc cctcgctcaa gccttcgtca ctggtcccgc caccaaacgt ttcggcgaga
 2221 agcaggccat tatcgccggc atggcggccc cacgggtgcg catgatcgtg ctcctgtcgt
 2281 tgaggacccg gctaggctgg cggggttgcc ttactggtta gcagaatgaa tcaccgatac
 2341 gcgagcgaac gtgaagcgac tgctgctgca aaacgtctgc gacctgagca acaacatgaa
 2401 tggtcttcgg tttccgtgtt tcgtaaagtc tggaaacgcg gaagtcagcg ccctgcacca
 2461 ttatgttccg gatctgcatc gcaggatgct gctggctacc ctgtggaaca cctacatctg
 2521 tattaacgaa gcgctggcat tgaccctgag tgatttttct ctggtcccgc cgcatccata
 2581 ccgccagttg tttaccctca caacgttcca gtaaccgggc atgttcatca tcagtaaccc
 2641 gtatcgtgag catcctctct cgtttcatcg gtatcattac ccccatgaac agaaatcccc
 2701 cttacacgga ggcatcagtg accaaacagg aaaaaaccgc ccttaacatg gcccgcttta
 2761 tcagaagcca gacattaacg cttctggaga aactcaacga gctggacgcg gatgaacagg
 2821 cagacatctg tgaatcgctt cacgaccacg ctgatgagct ttaccgcagc tgcctcgcgc
 2881 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
 2941 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
 3001 ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa
 3061 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atatgcggtg tgaaataccg
 3121 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
 3181 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
 3241 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
 3301 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
 3361 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
 3421 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
 3481 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
 3541 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
 3601 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
 3661 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
 3721 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
 3781 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
 3841 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
 3901 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
 3961 gctcagtgga acgaaaactc acgttaaggg attttggtca tgaacaataa aactgtctgc
 4021 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcttgctc
 4081 taggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
 4141 taatgtcggg caatcaggtg cgacaatcta tcgattgtat gggaagcccg atgcgccaga
 4201 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
 4261 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
 4321 tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc aggtattaga
 4381 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
 4441 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
 4501 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
 4561 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
 4621 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
 4681 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
 4741 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
 4801 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
 4861 ctaagaatta attcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
 4921 gggttccgcg cacatttccc cgaaaagtgc cacctgaaat tgtaaacgtt aatattttgt
 4981 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
 5041 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
 5101 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
 5161 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
 5221 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
 5281 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
 5341 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
 5401 tacagggcgc gtcccattcg cca
//

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