pET-30c(+)載體基本信息
別名: | pET30c, pet 30c |
---|---|
質(zhì)粒類(lèi)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: | 5423 bp |
5' 測(cè)序引物: | T7 |
5' 測(cè)序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
載體標(biāo)簽: | N-His, N-S, N-Thrombin, N-EK, C-His |
載體抗性: | Kanamycin |
表達(dá)宿主菌: | BL21(DE3) 、 Rosetta2(DE3)、C43(DE3) |
備注: | N term thrombin cleavage site; N term enterokinase cleavage site; a,b,c vary by MCS |
產(chǎn)品目錄號(hào): | 69911-3 |
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 | 129 | 1 | |
T7_Terminal_primer | 69 | 87 | |
EK | 231 | 220 | |
lacI | 303 | 1909 | |
T7_transl_en_RBS | 373 | 357 | |
lacO | 418 | 391 | |
T7_promoter | 436 | 418 | |
tet (300 - 563) | 472 | 735 | |
pBRrevBam_primer | 543 | 524 | |
tet (576 - 851) | 1968 | 2243 | |
ROP | 2718 | 2909 | |
pGEX_3_primer | 2925 | 2903 | |
pBR322_origin | 3943 | 3324 | |
KanR2 | 4049 | 4864 |
ORF | Start | End | |
---|---|---|---|
ORF frame 1 | 328 | 1077 | |
ORF frame 2 | 950 | 1909 | |
ORF frame 2 | 4049 | 4864 |
Enzyme Name | Cut |
---|---|
XhoI | 158 |
NotI | 166 |
EagI | 166 |
HindIII | 173 |
SalI | 179 |
SacI | 190 |
EcoRI | 192 |
BamHI | 198 |
EcoRV | 209 |
NcoI | 213 |
KpnI | 239 |
BglII | 242 |
BstBI | 269 |
NdeI | 347 |
XbaI | 385 |
BclI | 1191 |
ApaI | 1388 |
HpaI | 1683 |
FspI | 2259 |
NruI | 4137 |
SmaI | 4354 |
XmaI | 4352 |
The pET-30a-c(+) vectors carry an N-terminal His? Tag®/ thrombin/ S? Tag?/ enterokinase configuration plus an optional C-terminal His? Tag sequence. 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 circular 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).