pET-42b(+)

pET-42b(+)载体基本信息

出品公司:	Ybscience
别名:	pET42b, pet 42b, pET-42b(+)
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:	5929bp
5' 测序引物:	T7
5' 测序引物序列:	5'-TAATACGACTCACTATAGGG-3'
载体标签:	N-GST, N-His, C-His
载体抗性:	Kanamycin
备注:	含有GST融合蛋白;N端Thrombin蛋白酶切位点; 含有N端Factor Xa蛋白酶切位点 ; pET 42 a,b,c 的差异仅在多克隆位点区域。
稳定性:	瞬时表达 Transient
组成型:	组成型 Constitutive
病毒/非病毒:	非病毒

pET-42b(+)载体质粒图谱和多克隆位点信息

Feature Name	Start	End
T7_terminator	120	1
T7_Terminal_primer	69	87
S15	350	306
6xHIS	410	393
GST (variant)	1091	441
T7_transl_en_RBS	1116	1100
lacO	1161	1134
T7_promoter	1179	1161
tet (300 - 563)	1215	1478
pBRrevBam_primer	1286	1267
lacI	1561	2652
ROP	3224	3415
pGEX_3_primer	3431	3409
pBR322_origin	4449	3830
KanR2	4555	5370

ORF	Start	End
ORF frame 3	1091	147
ORF frame 3	1092	1820
ORF frame 1	1693	2652
ORF frame 1	4555	5370

Enzyme Name	Cut
XhoI	174
NotI	182
EagI	182
HindIII	189
SalI	195
SacI	206
PstI	214
AscI	216
StuI	226
EcoRI	235
BamHI	241
EcoRV	251
NcoI	254
AgeI	288
KpnI	295
BglII	298
SacII	389
SpeI	420
MscI	883
NdeI	1090
XbaI	1128
ApaI	2131
HpaI	2426
NruI	4643
SmaI	4860
XmaI	4858

 

pET-42b(+)载体简介

The pET-42 series is designed for cloning and high-level expression of peptide sequencesfused with the 220 aa GST•Tag™ protein. Unique sites are shown on the circle map. Note that thesequence is numbered by the pBR322 convention, so the T7 expression region is reversed on thecircle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymeraseis shown below. The f1 origin is oriented so that infection with helper phage will produce virionscontaining single stranded DNA that corresponds to the coding strand. Therefore, single strandedsequencing should be performed using the T7 terminator primer (cat. no. 69337-3). Vector encodedsequence can be completely removed when cloning into the PshAI site (as shown below) and thencleaving the GST fusion protein with Factor Xa.

pet42b载体序列

LOCUS       pET-42b(+)    5929 bp     DNA  circular   SYNDEFINITION  pET-42b(+), pet42bACCESSION   KEYWORDS    SOURCE        ORGANISM  other sequences; artificial sequences; vectors.COMMENT     This file is created by Vector NTI                            http://www.biofeng.com/COMMENT     ORIGDB|GenBankCOMMENT     VNTAUTHORNAME|Biofeng.com|COMMENT     VNTOAUTHORAD1|Novagen Inc.|FEATURES             Location/Qualifiers     source          1..5929                     /organism="pET-42 b(+)"                     /mol_type="other DNA"     terminator      complement(1..120)                     /label="T7_terminator"     misc_feature    69..87                     /label="T7_Terminal_primer"     CDS             complement(147..1091)                     /label="ORF frame 3"     misc_feature    complement(306..350)                     /label="S15"     misc_feature    complement(393..410)                     /label="6xHIS"     gene            complement(441..1091)                     /label="GST (variant)"                     /gene="GST (variant)"     CDS             1092..1820                     /label="ORF frame 3"     misc_feature    complement(1100..1116)                     /label="T7_transl_en_RBS"     misc_feature    complement(1134..1161)                     /label="lacO"     promoter        complement(1161..1179)                     /label="T7_promoter"     gene            1215..1478                     /label="tet (300 - 563)"                     /gene="tet (300 - 563)"     misc_feature    complement(1267..1286)                     /label="pBRrevBam_primer"     misc_feature    1561..2652                     /label="lacI"     CDS             1693..2652                     /label="ORF frame 1"     misc_feature    3224..3415                     /label="ROP"     misc_feature    complement(3409..3431)                     /label="pGEX_3_primer"     rep_origin      complement(3830..4449)                     /label="pBR322_origin"     gene            4555..5370                     /label="KanR2"                     /gene="KanR2"     CDS             4555..5370                     /label="ORF frame 1"ORIGIN    1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa   61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt  121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt  181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc  241 ggatccccga tatccatgga cccgcgtccc tcaataccgg agccaccacc ggtacccaga  301 tctgggctgt ccatgtgctg gcgttcgaat ttagcagcag cggtttcttt cataccaatt  361 gcagtactac cgcgtggcac cagacccgcg gagtgatggt gatggtgatg accagaacca  421 ctagttgaac catccgattt tggaggatgg tcgccaccac caaacgtggc ttgccagccc  481 tgcaaaggcc atgctatata cttgctggat ttcaagtact tatcaatttg tgggatagct  541 tcaatacgtt ttttaaaaca aactaatttt gggaacgcat ccaggcacat tgggtccatg  601 tataaaacaa catcaagagc gtcatacaac atgaagtcag gatgggttac atgatcacca  661 tttaaatatg ttttatgaca taaacgatct tcgaacattt tcagcatttc aggtagcttg  721 ctaagaaaat caactttgag agtttcaaag tctttactat atgcaattct cgaaacaccg  781 tatctaatat ccaaaaccgc tccttcaagc attgaaatct ctgcacgctc ttttggacaa  841 ccacccaaca tgttgtgctt gtcagctata taacgtatga tggccataga ctgtgttaat  901 ttaacatcac catcaatata ataaggaaga ttgggaaact ccaaacccaa ttcaaacttt  961 ttgtttcgcc atttatcacc ttcatcgcgc tcatacaaat gctcttcata tttttcttca 1021 agatattcca aaagaagtcg agtgggttgc acaaggccct taattttcca ataacctagt 1081 ataggggaca tatgtatatc tccttcttaa agttaaacaa aattatttct agaggggaat 1141 tgttatccgc tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatcga 1201 tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg 1261 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca 1321 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg 1381 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac 1441 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc 1501 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg 1561 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat 1621 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa 1681 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg 1741 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg 1801 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg 1861 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa 1921 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa 1981 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac 2041 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg 2101 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt 2161 ctggctggct ggcataaata tctcactcgc aatcaaattc agccgatagc ggaacgggaa 2221 ggcgactgga gtgccatgtc cggttttcaa caaaccatgc aaatgctgaa tgagggcatc 2281 gttcccactg cgatgctggt tgccaacgat cagatggcgc tgggcgcaat gcgcgccatt 2341 accgagtccg ggctgcgcgt tggtgcggac atctcggtag tgggatacga cgataccgaa 2401 gacagctcat gttatatccc gccgttaacc accatcaaac aggattttcg cctgctgggg 2461 caaaccagcg tggaccgctt gctgcaactc tctcagggcc aggcggtgaa gggcaatcag 2521 ctgttgcccg tctcactggt gaaaagaaaa accaccctgg cgcccaatac gcaaaccgcc 2581 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa 2641 agcgggcagt gagcgcaacg caattaatgt aagttagctc actcattagg caccgggatc 2701 tcgaccgatg cccttgagag ccttcaaccc agtcagctcc ttccggtggg cgcggggcat 2761 gactagcatg atcgtgctcc tgtcgttgag gacccggcta ggctggcggg gttgccttac 2821 tggttagcag aatgaatcac cgatacgcga gcgaacgtga agcgactgct gctgcaaaac 2881 gtctgcgacc tgagcaacaa catgaatggt cttcggtttc cgtgtttcgt aaagtctgga 2941 aacgcggaag tcagcgccct gcaccattat gttccggatc tgcatcgcag gatgctgctg 3001 gctaccctgt ggaacaccta catctgtatt aacgaagcgc tggcattgac cctgagtgat 3061 ttttctctgg tcccgccgca tccataccgc cagttgttta ccctcacaac gttccagtaa 3121 ccgggcatgt tcatcatcag taacccgtat cgtgagcatc ctctctcgtt tcatcggtat 3181 cattaccccc atgaacagaa atccccctta cacggaggca tcagtgacca aacaggaaaa 3241 aaccgccctt aacatggccc gctttatcag aagccagaca ttaacgcttc tggagaaact 3301 caacgagctg gacgcggatg aacaggcaga catctgtgaa tcgcttcacg accacgctga 3361 tgagctttac cgcagctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat 3421 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg 3481 tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca gccatgaccc agtcacgtag 3541 cgatagcgga gtgtatactg gcttaactat gcggcatcag agcagattgt actgagagtg 3601 caccatatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc 3661 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg 3721 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa 3781 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg 3841 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga 3901 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg 3961 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg 4021 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc 4081 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg 4141 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca 4201 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt 4261 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag 4321 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg 4381 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc 4441 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt 4501 tggtcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc 4561 catattcaac gggaaacgtc ttgctctagg ccgcgattaa attccaacat ggatgctgat 4621 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcga 4681 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc 4741 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg 4801 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc 4861 gggaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat 4921 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac 4981 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat 5041 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg 5101 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat 5161 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc 5221 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca 5281 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag 5341 tttcatttga tgctcgatga gtttttctaa gaattaattc atgagcggat acatatttga 5401 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc 5461 tgaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 5521 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 5581 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 5641 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 5701 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 5761 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 5821 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 5881 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgcca//