pET-40b(+)載體基本信息
別名: | pET40b, pET 40b |
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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: | 6190 bp |
5' 測序引物: | T7 |
5' 測序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
載體標(biāo)簽: | His (中間和C端), N-DsbC, N-S, N-EK, N-Thrombin |
載體抗性: | Kanamycin |
備注: |
Encodes DsbC tag for export and periplasmic folding of target proteins.
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表達(dá)宿主菌: | BL21(DE3) 、 Rosetta2(DE3)、C43(DE3) |
產(chǎn)品目錄號: | 70091-3 |
穩(wěn)定性: | 瞬時表達(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 | 266 | 252 | |
S15 | 326 | 282 | |
6xHIS | 386 | 369 | |
T7_transl_en_RBS | 1140 | 1124 | |
lacO | 1185 | 1158 | |
T7_promoter | 1203 | 1185 | |
tet (300 - 563) | 1239 | 1502 | |
pBRrevBam_primer | 1310 | 1291 | |
lacI | 1585 | 2676 | |
tet (576 - 851) | 2735 | 3010 | |
ROP | 3485 | 3676 | |
pGEX_3_primer | 3692 | 3670 | |
pBR322_origin | 4710 | 4091 | |
KanR2 | 4816 | 5631 |
ORF | Start | End | |
---|---|---|---|
ORF frame 3 | 1115 | 147 | |
ORF frame 3 | 1116 | 1844 | |
ORF frame 1 | 1717 | 2676 | |
ORF frame 1 | 4816 | 5631 |
Enzyme Name | Cut |
---|---|
XhoI | 174 |
NotI | 182 |
EagI | 182 |
HindIII | 189 |
SalI | 195 |
SacI | 206 |
EcoRI | 208 |
BamHI | 214 |
EcoRV | 222 |
NcoI | 227 |
KpnI | 271 |
BglII | 274 |
BstBI | 301 |
SacII | 365 |
SpeI | 396 |
NdeI | 1114 |
XbaI | 1152 |
BclI | 1958 |
ApaI | 2155 |
HpaI | 2450 |
FspI | 3026 |
NruI | 4904 |
The pET-40b(+) vector (Cat. No. 70091-3) is designed for expression of DsbC fusion proteins. 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).