pET-49b(+)

价格:800元

联系方式:I47-825O-882O

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pET49b载体基本信息

出品公司: EMD Biosciences (Novagen)
别名: pET49b, pET 49b
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: 5926 bp
5' 测序引物: T7或者pGEX 5'
5' 测序引物序列:   T7: 5'-TAATACGACTCACTATAGGG-3'; pGEX 5': 5'-GGGCTGGCAAGCCACGTTTGGTG-3'
3' 测序引物: T7t
3' 测序引物序列:   T7t: 5'-TGCTAGTTATTGCTCAGCGG-3'
载体标签: N-GST, N-His, N-HRV 3C, C-S, C-Thrombin
载体抗性: Kanamycin
表达宿主菌: BL21(DE3) Rosetta2(DE3)C43(DE3)
备注:
Same as pET47 but also has Nterm GST Tag; contains HRV 3C Protease cleavage site for fusion tag removal at low temperatures; Cterm thrombin cleavage site.
产品目录号: 71463-3
稳定性: 瞬时表达 Transient
组成型: 组成型 Constitutive
病毒/非病毒: 非病毒

pET49b载体质粒图谱和多克隆位点信息

pET-49b载体图谱



pET-49b多克隆位点

pET-49b载体特征

Feature Name Start End  
T7_terminator 120 1
T7_Terminal_primer 69 87
S15 212 168
6xHIS 359 342
GST (variant) 1088 438
T7_transl_en_RBS 1113 1097
lacO 1158 1131
T7_promoter 1176 1158
tet (300 - 563) 1212 1475
pBRrevBam_primer 1283 1264
lacI 1558 2649
ROP 3221 3412
pGEX_3_primer 3428 3406
pBR322_origin 4446 3827
KanR2 4552 5367
f1_origin 5898 5592
ORF Start End  
ORF frame 3 1088 141
ORF frame 3 1089 1817
ORF frame 1 1690 2649
ORF frame 1 4552 5367
Enzyme Name Cut
PacI 141
XhoI 147
SacI 238
NotI 242
EagI 242
HindIII 249
SalI 255
AatII 266
AscI 270
StuI 280
EcoRI 289
BamHI 295
KpnI 306
XmaI 306
SmaI 308
SacII 338
SpeI 417
MscI 880
NdeI 1087
XbaI 1125
ApaI 2128
HpaI 2423
NruI 4640

pET49b载体简介

The pET-49b(+) vector carries N-terminal GST•Tag™ and His•Tag? coding sequences followed by a recognition site for the human rhinovirus (HRV) 3C protease. This protease is highly specifi c for cleavage of the sequence LEVLFQ↓GP , and is active at low temperatures . pET-49b(+) also contains an optional C-terminal thrombin recognition site followed by an S•Tag™ coding sequence. Unique restriction .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 the helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the AS S•Tag 18mer Primer (Cat. No. 71262-3). 
pET Vectors 选择指南

pET49b载体序列

LOCUS       pET-49b(+)              5926 bp    DNA     circular     11-SEP-2008
DEFINITION  pET-49b(+).
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.|
COMMENT     VNTREPLTYPE|Plasmid
COMMENT     VNTEXTCHREPL|Bacteria
FEATURES             Location/Qualifiers
     rep_origin      complement(5467..5914)
                     /vntifkey="33"
                     /label=f1\ori
     misc_RNA        complement(3843..4398)
                     /vntifkey="53"
                     /label=RNA\II
     rep_origin      complement(3843..3843)
                     /vntifkey="33"
                     /label=ColE1,\pBR\ori
     misc_RNA        4283..4390
                     /vntifkey="53"
                     /label=RNA\I
     CDS             3221..3412
                     /vntifkey="4"
                     /label=ROP\gene
     CDS             1567..2649
                     /vntifkey="4"
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(1160..1176)
                     /vntifkey="30"
                     /label=T7\promoter
     protein_bind    complement(1135..1155)
                     /vntifkey="31"
                     /label=lacO
     CDS             complement(429..1088)
                     /vntifkey="4"
                     /label=GST
     protein_bind    complement(342..359)
                     /vntifkey="31"
                     /label=His-Tag
     terminator      complement(26..73)
                     /vntifkey="43"
                     /label=T7\terminator
     CDS             4552..5367
                     /vntifkey="4"
                     /label=Kan
     misc_feature    complement(213..230)
                     /vntifkey="21"
                     /label=Thrombin\site
     protein_bind    complement(168..212)
                     /vntifkey="31"
                     /label=S-Tag
     protein_bind    complement(310..332)
                     /vntifkey="31"
                     /label=3c
     primer          complement(300..336)
                     /vntifkey="27"
                     /label=S-SacII/Bam\+SanDI\-dcm
     primer          complement(374..421)
                     /vntifkey="27"
                     /label=S-Tag\Spacer-Spe\I
BASE COUNT     1485 a      1486 c      1520 g      1435 t
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgtttagcag cctaggttaa ttaagcctcg agagcagcag aagtagagct gtccatgtgc
      181 tggcgttcga atttagcagc agcggtttct ttactaccgc gtggcaccag agcgagctct
      241 gcggccgcaa gcttgtcgac ggacgtcggg cgcgccaagg cctgtacaga attcggatcc
      301 tggtacccgg gtccctgaaa gaggacttca agagccgcgg agtgatggtg atggtgatga
      361 ccagaaccac tagatggagt aggagtagga ggattgttat tagaaccgcc accaccacta
      421 gttgaaccat ccgattttgg aggatggtcg ccaccaccaa acgtggcttg ccagccctgc
      481 aaaggccatg ctatatactt gctggatttc aagtacttat caatttgtgg gatagcttca
      541 atacgttttt taaaacaaac taattttggg aacgcatcca ggcacattgg gtccatgtat
      601 aaaacaacat caagagcgtc atacaacatg aagtcaggat gggttacatg atcaccattt
      661 aaatatgttt tatgacataa acgatcttcg aacattttca gcatttcagg tagcttgcta
      721 agaaaatcaa ctttgagagt ttcaaagtct ttactatatg caattctcga aacaccgtat
      781 ctaatatcca aaaccgctcc ttcaagcatt gaaatctctg cacgctcttt tggacaacca
      841 cccaacatgt tgtgcttgtc agctatataa cgtatgatgg ccatagactg tgttaattta
      901 acatcaccat caatataata aggaagattg ggaaactcca aacccaattc aaactttttg
      961 tttcgccatt tatcaccttc atcgcgctca tacaaatgct cttcatattt ttcttcaaga
     1021 tattccaaaa gaagtcgagt gggttgcaca aggcccttaa ttttccaata acctagtata
     1081 ggggacatat gtatatctcc ttcttaaagt taaacaaaat tatttctaga ggggaattgt
     1141 tatccgctca caattcccct atagtgagtc gtattaattt cgcgggatcg agatcgatct
     1201 cgatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg
     1261 gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga
     1321 gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca
     1381 tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac ctactactgg
     1441 gctgcttcct aatgcaggag tcgcataagg gagagcgtcg agatcccgga caccatcgaa
     1501 tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca attcagggtg
     1561 gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt ctcttatcag
     1621 accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg ggaaaaagtg
     1681 gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca actggcgggc
     1741 aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc gccgtcgcaa
     1801 attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt ggtgtcgatg
     1861 gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct cgcgcaacgc
     1921 gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc tgtggaagct
     1981 gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc catcaacagt
     2041 attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt cgcattgggt
     2101 caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg tctgcgtctg
     2161 gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga acgggaaggc
     2221 gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga gggcatcgtt
     2281 cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg cgccattacc
     2341 gagtccgggc tgcgcgttgg tgcggacatc tcggtagtgg gatacgacga taccgaagac
     2401 agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct gctggggcaa
     2461 accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg caatcagctg
     2521 ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca aaccgcctct
     2581 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2641 gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac cgggatctcg
     2701 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     2761 tagcatgatc gtgctcctgt cgttgaggac ccggctaggc tggcggggtt gccttactgg
     2821 ttagcagaat gaatcaccga tacgcgagcg aacgtgaagc gactgctgct gcaaaacgtc
     2881 tgcgacctga gcaacaacat gaatggtctt cggtttccgt gtttcgtaaa gtctggaaac
     2941 gcggaagtca gcgccctgca ccattatgtt ccggatctgc atcgcaggat gctgctggct
     3001 accctgtgga acacctacat ctgtattaac gaagcgctgg cattgaccct gagtgatttt
     3061 tctctggtcc cgccgcatcc ataccgccag ttgtttaccc tcacaacgtt ccagtaaccg
     3121 ggcatgttca tcatcagtaa cccgtatcgt gagcatcctc tctcgtttca tcggtatcat
     3181 tacccccatg aacagaaatc ccccttacac ggaggcatca gtgaccaaac aggaaaaaac
     3241 cgcccttaac atggcccgct ttatcagaag ccagacatta acgcttctgg agaaactcaa
     3301 cgagctggac gcggatgaac aggcagacat ctgtgaatcg cttcacgacc acgctgatga
     3361 gctttaccgc agctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     3421 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     3481 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga
     3541 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac
     3601 catatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgct
     3661 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     3721 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     3781 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     3841 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     3901 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     3961 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4021 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4081 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4141 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4201 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4261 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4321 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4381 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4441 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4501 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     4561 attcaacggg aaacgtcttg ctctaggccg cgattaaatt ccaacatgga tgctgattta
     4621 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgattg
     4681 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     4741 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     4801 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccggc
     4861 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     4921 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagt
     4981 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     5041 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     5101 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     5161 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     5221 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     5281 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     5341 catttgatgc tcgatgagtt tttctaagaa ttaattcatg agcggataca tatttgaatg
     5401 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     5461 aattgtaaac gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     5521 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     5581 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     5641 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     5701 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     5761 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     5821 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     5881 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgcca
//

pET49b载体质粒图谱pdf版和相关资料下载

pET49b载体应用举例

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