IV style="POSITION: relative; TEXT-ALIGN: left; PADDING-BOTTOM: 0px; TEXT-TRANSFORM: none; BACKGROUND-COLOR: rgb(255,255,255); TEXT-INDENT: 0px; MARGIN: 0px 0px 16px; PADDING-LEFT: 0px; LETTER-SPACING: normal; PADDING-RIGHT: 0px; FONT: 14px Verdana, Arial, Tahoma; MAX-WIDTH: 620px; WHITE-SPACE: normal; COLOR: rgb(51,51,51); WORD-SPACING: 0px; PADDING-TOP: 0px; -webkit-text-stroke-width: 0px" class=main-title>
pET32b载体基本信息
出品公司:别名:质粒类型:表达水平:克隆方法:载体大小:5 测序引物:5 测序引物序列:  载体标签:载体抗性:备注:产品目录号:稳定性:组成型:病毒/非病毒:
EMD Biosciences(Novagen)
pET32b, pet 32b
大肠杆菌蛋白表达
高
多克隆位点，限制性内切酶
5899bp
T7或者Trx-F
T7: 5-TAATACGACTCACTATAGGG-3; Trx-F: 5 TTCCTCGACGCTAACCTG 3
thioredoxin (N端); His (中间和C端)
Ampicillin
Production of soluble, active target proteins; N-term thrombin cleavage site;
Nterm enterokinase cleavage site; a,b,c vary by MCS
69016-3
瞬时表达 Transient
组成型 Constitutive
非病毒
pET32b载体质粒图谱和多克隆位点信息

pET32b载体简介
The pET-32a-c series is designed for cloning and high-level expression of peptide sequences fused with the 109aa Trx•Tag™ thioredoxin protein (1). Cloning sites are available for producing fusion proteins also containing cleavable His•Tag® and S•Tag™ sequences for detection and purification. 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 circle 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).
pET32b载体序列
LOCUS       pET-32b(+)              5899 bp    DNA     circular     11-SEP-2008
SOURCE      
  ORGANISM  
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
     promoter        complement(763..779)
                     /vntifkey="30"
                     /label=T7\promoter
     misc_signal     complement(365..691)
                     /vntifkey="87"
                     /label=trx\tag
     protein_bind    complement(326..343)
                     /vntifkey="31"
                     /label=His\tag
     misc_feature    complement(248..292)
                     /vntifkey="21"
                     /label=S\tag
     protein_bind    complement(140..157)
                     /vntifkey="31"
                     /label=His\tag
     terminator      complement(26..72)
                     /vntifkey="43"
                     /label=T7\terminator
     CDS             1170..2249
                     /vntifkey="4"
                     /label=lac\I
     rep_origin      complement(3683..3683)
                     /vntifkey="33"
                     /label=ColE1\pBR322\origin
     CDS             complement(736..760)
                     /vntifkey="4"
                     /label=lac\operator
     protein_bind    complement(299..316)
                     /vntifkey="31"
                     /label=thrombin
     protein_bind    complement(218..232)
                     /vntifkey="31"
                     /label=enterokinase
     CDS             complement(4444..5301)
                     /vntifkey="4"
                     /label=bla\(amp)\sequence
     rep_origin      complement(5433..5888)
                     /vntifkey="33"
                     /label=f1\origin
BASE COUNT     1390 a      1564 c      1560 g      1385 t
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 tccgatatcg ccatggcctt gtcgtcgtcg tcggtaccca
      241 gatctgggct gtccatgtgc tggcgttcga atttagcagc agcggtttct ttcataccag
      301 aaccgcgtgg caccagacca gaagaatgat gatgatgatg gtgcatatgg ccagaaccag
      361 aaccggccag gttagcgtcg aggaactctt tcaactgacc tttagacagt gcacccactt
      421 tggttgccgc cacttcaccg tttttgaaca gcagcagagt cgggatacca cggatgccat
      481 atttcggcgc agtgccaggg ttttgatcga tgttcagttt tgcaacggtc agtttgccct
      541 gatattcgtc agcgatttca tccagaatcg gggcgatcat tttgcacgga ccgcaccact
      601 ctgcccagaa atcgacgagg atcgccccgt ccgctttgag tacatccgtg tcaaaactgt
      661 cgtcagtcag gtgaataatt ttatcgctca tatgtatatc tccttcttaa agttaaacaa
      721 aattatttct agaggggaat tgttatccgc tcacaattcc cctatagtga gtcgtattaa
      781 tttcgcggga tcgagatcga tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac
      841 cggcgccaca ggtgcggttg ctggcgccta tatcgccgac atcaccgatg gggaagatcg
      901 ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt
      961 ggccggggga ctgttgggcg ccatctcctt gcatgcacca ttccttgcgg cggcggtgct
     1021 caacggcctc aacctactac tgggctgctt cctaatgcag gagtcgcata agggagagcg
     1081 tcgagatccc ggacaccatc gaatggcgca aaacctttcg cggtatggca tgatagcgcc
     1141 cggaagagag tcaattcagg gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag
     1201 agtatgccgg tgtctcttat cagaccgttt cccgcgtggt gaaccaggcc agccacgttt
     1261 ctgcgaaaac gcgggaaaaa gtggaagcgg cgatggcgga gctgaattac attcccaacc
     1321 gcgtggcaca acaactggcg ggcaaacagt cgttgctgat tggcgttgcc acctccagtc
     1381 tggccctgca cgcgccgtcg caaattgtcg cggcgattaa atctcgcgcc gatcaactgg
     1441 gtgccagcgt ggtggtgtcg atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg
     1501 tgcacaatct tctcgcgcaa cgcgtcagtg ggctgatcat taactatccg ctggatgacc
     1561 aggatgccat tgctgtggaa gctgcctgca ctaatgttcc ggcgttattt cttgatgtct
     1621 ctgaccagac acccatcaac agtattattt tctcccatga agacggtacg cgactgggcg
     1681 tggagcatct ggtcgcattg ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt
     1741 ctgtctcggc gcgtctgcgt ctggctggct ggcataaata tctcactcgc aatcaaattc
     1801 agccgatagc ggaacgggaa ggcgactgga gtgccatgtc cggttttcaa caaaccatgc
     1861 aaatgctgaa tgagggcatc gttcccactg cgatgctggt tgccaacgat cagatggcgc