pET-41b(+)載體基本信息
別名: | pET41b, pET 41b |
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質(zhì)粒類型: | 大腸桿菌蛋白表達(dá) |
表達(dá)水平: | 高 |
克隆方法: | 多克隆位點(diǎn),限制性內(nèi)切酶 |
載體大小: | 5932 bp |
5' 測(cè)序引物: | T7或pGEX-5‘ |
5' 測(cè)序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3'; pGEX-5': 5'-GGGCTGGCAAGCCACGTTTGGTG-3' |
載體標(biāo)簽: | N-GST, N-His, N-Thrombin |
載體抗性: | Kanamycin |
表達(dá)宿主菌: | BL21(DE3) 、 Rosetta2(DE3)、C43(DE3) |
備注: |
Encodes GST fusion tag; Nterm thrombin cleavage site; Nterm enterokinase cleavage site; PshAI blunt cloning site; a,b,c vary by MCS
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產(chǎn)品目錄號(hào): | 70557-3 |
穩(wěn)定性: | 瞬時(shí)表達(dá) Transient |
組成型: | 組成型 Constitutive |
病毒/非病毒: | 非病毒 |
Feature Name | Start | End | |
---|---|---|---|
T7_terminator | 120 | 1 | |
T7_Terminal_primer | 69 | 87 | |
EK | 278 | 264 | |
S15 | 353 | 309 | |
6xHIS | 413 | 396 | |
GST (variant) | 1094 | 444 | |
T7_transl_en_RBS | 1119 | 1103 | |
lacO | 1164 | 1137 | |
T7_promoter | 1182 | 1164 | |
tet (300 - 563) | 1218 | 1481 | |
pBRrevBam_primer | 1289 | 1270 | |
lacI | 1564 | 2655 | |
ROP | 3227 | 3418 | |
pGEX_3_primer | 3434 | 3412 | |
pBR322_origin | 4452 | 3833 | |
KanR2 | 4558 | 5373 |
ORF | Start | End | |
---|---|---|---|
ORF frame 3 | 1094 | 147 | |
ORF frame 3 | 1095 | 1823 | |
ORF frame 1 | 1696 | 2655 | |
ORF frame 1 | 4558 | 5373 |
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 | 291 |
KpnI | 298 |
BglII | 301 |
SacII | 392 |
SpeI | 423 |
MscI | 886 |
NdeI | 1093 |
XbaI | 1131 |
ApaI | 2134 |
HpaI | 2429 |
NruI | 4646 |
SmaI | 4863 |
XmaI | 4861 |
The pET-41 series is designed for cloning and high-level expression of peptide sequences fused with the 220 aa GST?Tag? protein. 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).Vector encoded sequence can be completely removed when cloning into the PshAI site (as shown below) and then cleaving the GST fusion protein with Enterokinase.