pET-24b(+)

pET-24b(+)

pET24b载体基本信息

出品公司:	Ybscience
别名:	pET24b, pet 24b
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:	5309 bp
5' 测序引物:	T7
5' 测序引物序列:	5'-TAATACGACTCACTATAGGG-3'
载体标签:	N-T7, C-His
载体抗性:	Kanamycin
备注:	Same as pET21a,b,c,d(+) but kanamycin resistance; a,b,c,d vary by MCS; The f1 origin is oriented so that infection with helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand.
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

pET24b载体质粒图谱和多克隆位点信息

The maps for pET-24b(+), pET-24c(+) and pET-24d(+) are the same as pET-24a(+) (shown) with the following exceptions: pET-24b(+) is a 5309bp plasmid; subtract 1bp from each site beyond BamH I at 198. pET-24c(+) is a 5308bp plasmid; subtract 2bp from each site beyond BamH I at 198. pET-24d(+) is a 5307bp plasmid; the BamH I site is in the same reading frame as in pET- 24c(+). An Nco I site is substituted for the Nde I site with a net 1bp deletion at position 238 of pET-24c(+). As a result, Nco I cuts pET24d(+) at 234, and Nhe I cuts at 229. For the rest of the sites, subtract 3bp from each site beyond position 239 in pET-24a(+). Nde I does not cut pET-24d(+). Note also that Sty I is not unique in pET-24d(+).
 

Feature Name	Start	End
T7_terminator	129	1
T7_Terminal_primer	69	87
T7_leader	238	206
Xpress_fwd_primer	239	221
T7_transl_en_RBS	263	247
lacO	308	281
T7_promoter	326	308
tet (300 - 563)	358	621
pBRrevBam_primer	429	410
lacI	704	1795
tet (576 - 851)	1854	2129
ROP	2604	2795
pGEX_3_primer	2811	2789
pBR322_origin	3829	3210
KanR2	3935	4750

ORF	Start	End
ORF frame 1	463	963
ORF frame 2	836	1795
ORF frame 2	3935	4750

Enzyme Name	Cut
XhoI	158
NotI	166
EagI	166
HindIII	173
SalI	179
SacI	190
EcoRI	192
BamHI	198
NheI	230
NdeI	237
XbaI	275
BglII	341
BclI	1077
ApaI	1274
EcoRV	1513
HpaI	1569
FspI	2145
NruI	4023
ClaI	4057
XmaI	4238
SmaI	4240

 

pET24b载体简介

The pET-24a-d(+) vectors carry an N-terminal T7•Tag® sequence plus an optional C-terminal His•Tag® sequence. These vectors differ from pET-21a-d(+) only by their selectable marker (kanamycin vs. ampicillin resistance). 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, singlestranded sequencing should be performed using the T7 terminator primer.

pET24b载体序列

LOCUS       pET-24 b (+)    5309 bp     DNA circular    SYN
DEFINITION  pET-24 b (+)
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..5309
                     /organism="pET-24 b (+)"
                     /mol_type="other DNA"
     terminator      complement(1..129)
                     /label="T7_terminator"
     misc_feature    69..87
                     /label="T7_Terminal_primer"
     misc_feature    complement(206..238)
                     /label="T7_leader"
     misc_feature    complement(221..239)
                     /label="Xpress_fwd_primer"
     misc_feature    complement(247..263)
                     /label="T7_transl_en_RBS"
     misc_feature    complement(281..308)
                     /label="lacO"
     promoter        complement(308..326)
                     /label="T7_promoter"
     gene            358..621
                     /label="tet (300 - 563)"
                     /gene="tet (300 - 563)"
     misc_feature    complement(410..429)
                     /label="pBRrevBam_primer"
     CDS             463..963
                     /label="ORF frame 1"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     misc_feature    704..1795
                     /label="lacI"
                     /translation="MSACFGVGMVAGPVAGGLLGAISLHAPFLAAAVLNGLNLLLGCF
                     LMQESHKGERRDPGHHRMAQNLSRYGMIAPGRESIQGGECETSNVIRCRRVCRCLLSD
                     RFPRGEPGQPRFCENAGKSGSGDGGAELHSQPRGTTTGGQTVVADWRCHLQSGPARAV
                     ANCRGD*"
     CDS             836..1795
                     /label="ORF frame 2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            1854..2129
                     /label="tet (576 - 851)"
                     /gene="tet (576 - 851)"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    2604..2795
                     /label="ROP"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     misc_feature    complement(2789..2811)
                     /label="pGEX_3_primer"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     rep_origin      complement(3210..3829)
                     /label="pBR322_origin"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     gene            3935..4750
                     /label="KanR2"
                     /gene="KanR2"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKS
                     RADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNV
                     PALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGW
                     HKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITE
                     SGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQ
                     LLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ*"
     CDS             3935..4750
                     /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 tcccgaccca tttgctgtcc accagtcatg ctagccatat
  241 gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt tatccgctca
  301 caattcccct atagtgagtc gtattaattt cgcgggatcg agatctcgat cctctacgcc
  361 ggacgcatcg tggccggcat caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc
  421 gacatcaccg atggggaaga tcgggctcgc cacttcgggc tcatgagcgc ttgtttcggc
  481 gtgggtatgg tggcaggccc cgtggccggg ggactgttgg gcgccatctc cttgcatgca
  541 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg
  601 caggagtcgc ataagggaga gcgtcgagat cccggacacc atcgaatggc gcaaaacctt
  661 tcgcggtatg gcatgatagc gcccggaaga gagtcaattc agggtggtga atgtgaaacc
  721 agtaacgtta tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt
  781 ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc
  841 ggagctgaat tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct
  901 gattggcgtt gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat
  961 taaatctcgc gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg
 1021 cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat
 1081 cattaactat ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt
 1141 tccggcgtta tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca
 1201 tgaagacggt acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc
 1261 gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa
 1321 atatctcact cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat
 1381 gtccggtttt caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct
 1441 ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg
 1501 cgttggtgcg gatatctcgg tagtgggata cgacgatacc gaagacagct catgttatat
 1561 cccgccgtta accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg
 1621 cttgctgcaa ctctctcagg gccaggcggt gaagggcaat cagctgttgc ccgtctcact
 1681 ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
 1741 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
 1801 acgcaattaa tgtaagttag ctcactcatt aggcaccggg atctcgaccg atgcccttga
 1861 gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc gtcgccgcac
 1921 ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg ctctgggtca
 1981 ttttcggcga ggaccgcttt cgctggagcg cgacgatgat cggcctgtcg cttgcggtat
 2041 tcggaatctt gcacgccctc gctcaagcct tcgtcactgg tcccgccacc aaacgtttcg
 2101 gcgagaagca ggccattatc gccggcatgg cggccccacg ggtgcgcatg atcgtgctcc
 2161 tgtcgttgag gacccggcta ggctggcggg gttgccttac tggttagcag aatgaatcac
 2221 cgatacgcga gcgaacgtga agcgactgct gctgcaaaac gtctgcgacc tgagcaacaa
 2281 catgaatggt cttcggtttc cgtgtttcgt aaagtctgga aacgcggaag tcagcgccct
 2341 gcaccattat gttccggatc tgcatcgcag gatgctgctg gctaccctgt ggaacaccta
 2401 catctgtatt aacgaagcgc tggcattgac cctgagtgat ttttctctgg tcccgccgca
 2461 tccataccgc cagttgttta ccctcacaac gttccagtaa ccgggcatgt tcatcatcag
 2521 taacccgtat cgtgagcatc ctctctcgtt tcatcggtat cattaccccc atgaacagaa
 2581 atccccctta cacggaggca tcagtgacca aacaggaaaa aaccgccctt aacatggccc
 2641 gctttatcag aagccagaca ttaacgcttc tggagaaact caacgagctg gacgcggatg
 2701 aacaggcaga catctgtgaa tcgcttcacg accacgctga tgagctttac cgcagctgcc
 2761 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
 2821 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
 2881 ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
 2941 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatat gcggtgtgaa
 3001 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
 3061 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
 3121 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
 3181 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
 3241 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
 3301 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
 3361 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
 3421 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
 3481 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
 3541 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
 3601 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
 3661 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
 3721 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
 3781 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
 3841 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgaa caataaaact
 3901 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
 3961 ttgctctagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc
 4021 tcgcgataat gtcgggcaat caggtgcgac aatctatcga ttgtatggga agcccgatgc
 4081 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat
 4141 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg
 4201 tactcctgat gatgcatggt tactcaccac tgcgatcccc gggaaaacag cattccaggt
 4261 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg
 4321 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct
 4381 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga
 4441 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc
 4501 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg
 4561 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct
 4621 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca
 4681 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga
 4741 gtttttctaa gaattaattc atgagcggat acatatttga atgtatttag aaaaataaac
 4801 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaaattgta aacgttaata
 4861 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
 4921 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
 4981 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
 5041 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
 5101 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
 5161 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
 5221 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
 5281 cgccgctaca gggcgcgtcc cattcgcca