pET-29a(+)

价格:800元

联系方式:I47-825O-882O

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pET29a载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET29a, pet 29a
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: 5371 bp
5' 测序引物: T7
5' 测序引物序列:   5'-TAATACGACTCACTATAGGG-3'
载体标签: N-S, N-Thrombin, C-His
载体抗性: Kanamycin
表达宿主菌: BL21(DE3) BLR(DE3)BL21(DE3)pLysS
备注:
N term thrombin cleavage site; a,b,c vary by MCS;
 
产品目录号: 69871-3
稳定性: 瞬时表达 Transient
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒

pET29a载体质粒图谱和多克隆位点信息

pet29a载体图谱
pet29a多克隆位点

Feature Name Start End  
T7_terminator 129 1
T7_Terminal_primer 69 87
lacI 218 1857
T7_transl_en_RBS 321 305
lacO 366 339
T7_promoter 384 366
tet (300 - 563) 420 683
pBRrevBam_primer 491 472
tet (576 - 851) 1916 2191
ROP 2666 2857
pGEX_3_primer 2873 2851
pBR322_origin 3891 3272
KanR2 3997 4812
ORF Start End  
ORF frame 3 297 1025
ORF frame 1 898 1857
ORF frame 1 3997 4812
Enzyme Name Cut
XhoI 158
NotI 166
EagI 166
HindIII 173
SalI 179
SacI 190
EcoRI 192
BamHI 198
EcoRV 206
NcoI 212
KpnI 238
BglII 241
BstBI 268
NdeI 295
XbaI 333
BclI 1139
ApaI 1336
HpaI 1631
FspI 2207
NruI 4085
SmaI 4302
XmaI 4300

pET29a载体简介

The pET-29a-c(+) vectors carry an N-terminal S•Tag™/thrombin 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).

pET29a载体序列

LOCUS       pET-29 a (+)    5371 bp     DNA  Circular   SYN
DEFINITION  pET-29 a (+)
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..5371
                     /organism="pET-29 a (+)"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    218..1857
                     /label="lacI"
     CDS             297..1025
                     /label="ORF frame 3"
                     /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                     IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                     PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                     ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                     TVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(305..321)
                     /label="T7_transl_en_RBS"
                     /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                     IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                     PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                     ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                     TVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(339..366)
                     /label="lacO"
                     /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                     IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                     PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                     ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                     TVVADWRCHLQSGPARAVANCRGD*"
     promoter        complement(366..384)
                     /label="T7_promoter"
                     /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                     IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                     PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                     ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                     TVVADWRCHLQSGPARAVANCRGD*"
     gene            420..683
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
                     /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                     IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                     PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                     ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                     TVVADWRCHLQSGPARAVANCRGD*"
     misc_feature    complement(472..491)
                     /label="pBRrevBam_primer"
                     /translation="MYISFLKLNKIISRGELLSAHNSPIVSRINFAGSRSISILYAGR
                     IVAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHA
                     PFLAAAVLNGLNLLLGCFLMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGEC
                     ETSNVIRCRRVCRCLLSDRFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQ
                     TVVADWRCHLQSGPARAVANCRGD*"
     CDS             898..1857
                     /label="ORF frame 1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            1916..2191
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    2666..2857
                     /label="ROP"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2851..2873)
                     /label="pGEX_3_primer"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      complement(3272..3891)
                     /label="pBR322_origin"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            3997..4812
                     /label="KanR2"
                     /gene="KanR2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             3997..4812
                     /label="ORF frame 1"
                     /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 tccgatatca gccatggaac cgcgtggcac cagggtaccc
  241 agatctgggc tgtccatgtg ctggcgttcg aatttagcag cagcggtttc tttcatatgt
  301 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
  361 attcccctat agtgagtcgt attaatttcg cgggatcgag atcgatctcg atcctctacg
  421 ccggacgcat cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg
  481 ccgacatcac cgatggggaa gatcgggctc gccacttcgg gctcatgagc gcttgtttcg
  541 gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt gggcgccatc tccttgcatg
  601 caccattcct tgcggcggcg gtgctcaacg gcctcaacct actactgggc tgcttcctaa
  661 tgcaggagtc gcataaggga gagcgtcgag atcccggaca ccatcgaatg gcgcaaaacc
  721 tttcgcggta tggcatgata gcgcccggaa gagagtcaat tcagggtggt gaatgtgaaa
  781 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
  841 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
  901 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
  961 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
 1021 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
 1081 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
 1141 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
 1201 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
 1261 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
 1321 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
 1381 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
 1441 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
 1501 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
 1561 cgcgttggtg cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
 1621 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
 1681 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
 1741 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
 1801 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
 1861 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
 1921 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
 1981 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
 2041 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
 2101 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
 2161 cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct
 2221 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
 2281 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
 2341 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
 2401 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
 2461 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
 2521 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
 2581 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
 2641 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
 2701 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
 2761 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
 2821 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
 2881 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
 2941 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
 3001 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg
 3061 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc
 3121 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
 3181 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
 3241 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
 3301 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
 3361 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
 3421 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
 3481 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
 3541 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
 3601 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
 3661 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
 3721 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
 3781 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
 3841 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
 3901 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa
 3961 ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg
 4021 tcttgctcta ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
 4081 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
 4141 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
 4201 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
 4261 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag
 4321 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
 4381 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
 4441 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
 4501 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
 4561 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
 4621 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
 4681 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
 4741 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
 4801 gagtttttct aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa
 4861 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa
 4921 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
 4981 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
 5041 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
 5101 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
 5161 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
 5221 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
 5281 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
 5341 tgcgccgcta cagggcgcgt cccattcgcc a
//

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