pET-24(+)

pET-24(+)

pET24+载体基本信息

出品公司:	Ybscience
别名:	pET24(+), pet 24(+)
质粒类型:	大肠杆菌蛋白表达
表达水平:	高
克隆方法:	多克隆位点，限制性内切酶
载体大小:	5236 bp
5' 测序引物:	T7
5' 测序引物序列:	5'-TAATACGACTCACTATAGGG-3'
载体标签:	C-His
载体抗性:	Kana
备注:	Designed for expression from bacterial translational signal within cloned insert (no ribosome binding site or ATG start codon).
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

pET24+载体质粒图谱和多克隆位点信息

Feature Name	Start	End
T7_terminator	129	1
T7_Terminal_primer	69	87
lacO	235	208
T7_promoter	253	235
tet (300 - 563)	285	548
pBRrevBam_primer	356	337
lacI	631	1722
tet (576 - 851)	1781	2056
ROP	2531	2722
pGEX_3_primer	2738	2716
pBR322_origin	3756	3137
KanR2	3862	4677

ORF	Start	End
ORF frame 3	390	890
ORF frame 1	763	1722
ORF frame 1	3862	4677

Enzyme Name	Cut
XhoI	158
NotI	166
EagI	166
HindIII	173
SalI	179
SacI	190
EcoRI	192
BamHI	198
BglII	268
BclI	1004
ApaI	1201
EcoRV	1440
HpaI	1496
FspI	2072
NruI	3950
ClaI	3984
XmaI	4165
SmaI	4167

 

pET24+载体简介

pET-24(+) (Cat. No. 69772-3) is a transcription vector designed for expression from bacterial translation signals carried within a cloned insert. It therefore lacks the ribosome binding site and ATG start codon present on the pET translation vectors. A C-terminal His•Tag® sequence is available. 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).

pET24+载体序列

LOCUS       pET-24(+)    5236 bp     DNA Circular    SYN
DEFINITION  pET-24(+)
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..5236
                     /organism="pET-24(+)"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    complement(208..235)
                     /label="lacO"
     promoter        complement(235..253)
                     /label="T7_promoter"
     gene            285..548
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(337..356)
                     /label="pBRrevBam_primer"
     CDS             390..890
                     /label="ORF frame 3"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     misc_feature    631..1722
                     /label="lacI"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     CDS             763..1722
                     /label="ORF frame 1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            1781..2056
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    2531..2722
                     /label="ROP"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2716..2738)
                     /label="pGEX_3_primer"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      complement(3137..3756)
                     /label="pBR322_origin"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            3862..4677
                     /label="KanR2"
                     /gene="KanR2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             3862..4677
                     /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 tcctagaggg gaattgttat ccgctcacaa ttcccctata
  241 gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga cgcatcgtgg
  301 ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg
  361 gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg
  421 caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg
  481 cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata
  541 agggagagcg tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca
  601 tgatagcgcc cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac
  661 gatgtcgcag agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc
  721 agccacgttt ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac
  781 attcccaacc gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc
  841 acctccagtc tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc
  901 gatcaactgg gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt
  961 aaagcggcgg tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg
 1021 ctggatgacc aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt
 1081 cttgatgtct ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg
 1141 cgactgggcg tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc
 1201 ccattaagtt ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc
 1261 aatcaaattc agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa
 1321 caaaccatgc aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat
 1381 cagatggcgc tgggcgcaat gcgcgccatt accgagtccg ggctgcgcgt tggtgcggat
 1441 atctcggtag tgggatacga cgataccgaa gacagctcat gttatatccc gccgttaacc
 1501 accatcaaac aggattttcg cctgctgggg caaaccagcg tggaccgctt gctgcaactc
 1561 tctcagggcc aggcggtgaa gggcaatcag ctgttgcccg tctcactggt gaaaagaaaa
 1621 accaccctgg cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
 1681 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
 1741 aagttagctc actcattagg caccgggatc tcgaccgatg cccttgagag ccttcaaccc
 1801 agtcagctcc ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt
 1861 ctttatcatg caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga
 1921 ccgctttcgc tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca
 1981 cgccctcgct caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc
 2041 cattatcgcc ggcatggcgg ccccacgggt gcgcatgatc gtgctcctgt cgttgaggac
 2101 ccggctaggc tggcggggtt gccttactgg ttagcagaat gaatcaccga tacgcgagcg
 2161 aacgtgaagc gactgctgct gcaaaacgtc tgcgacctga gcaacaacat gaatggtctt
 2221 cggtttccgt gtttcgtaaa gtctggaaac gcggaagtca gcgccctgca ccattatgtt
 2281 ccggatctgc atcgcaggat gctgctggct accctgtgga acacctacat ctgtattaac
 2341 gaagcgctgg cattgaccct gagtgatttt tctctggtcc cgccgcatcc ataccgccag
 2401 ttgtttaccc tcacaacgtt ccagtaaccg ggcatgttca tcatcagtaa cccgtatcgt
 2461 gagcatcctc tctcgtttca tcggtatcat tacccccatg aacagaaatc ccccttacac
 2521 ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct ttatcagaag
 2581 ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac aggcagacat
 2641 ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg cgcgtttcgg
 2701 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
 2761 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
 2821 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct taactatgcg
 2881 gcatcagagc agattgtact gagagtgcac catatatgcg gtgtgaaata ccgcacagat
 2941 gcgtaaggag aaaataccgc atcaggcgct cttccgcttc ctcgctcact gactcgctgc
 3001 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
 3061 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
 3121 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
 3181 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
 3241 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
 3301 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
 3361 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
 3421 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
 3481 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
 3541 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
 3601 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
 3661 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
 3721 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
 3781 ggaacgaaaa ctcacgttaa gggattttgg tcatgaacaa taaaactgtc tgcttacata
 3841 aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcttg ctctaggccg
 3901 cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg cgataatgtc
 3961 gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc agagttgttt
 4021 ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt cagactaaac
 4081 tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac tcctgatgat
 4141 gcatggttac tcaccactgc gatccccggg aaaacagcat tccaggtatt agaagaatat
 4201 cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg gttgcattcg
 4261 attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc tcaggcgcaa
 4321 tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg taatggctgg
 4381 cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc ggattcagtc
 4441 gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa attaataggt
 4501 tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc catcctatgg
 4561 aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa atatggtatt
 4621 gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt tttctaagaa
 4681 ttaattcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
 4741 gcgcacattt ccccgaaaag tgccacctga aattgtaaac gttaatattt tgttaaaatt
 4801 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
 4861 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
 4921 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
 4981 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
 5041 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
 5101 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
 5161 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
 5221 cgcgtcccat tcgcca