pET-43.1a(+)載體基本信息
別名: | pET43.1a, pET 43.1a |
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質粒類型: | 大腸桿菌蛋白表達 |
表達水平: | 高 |
克隆方法: | 多克隆位點,限制性內切酶 |
載體大小: | 7275 bp |
5' 測序引物: | T7 |
5' 測序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
3' 測序引物: | ColiDOWN |
3' 測序引物序列: | ColiDOWN: 5'-TTCACTTCTGAGTTCGGCATG-3' |
載體標簽: | C-HSV, N-His, C-His, N-Thrombin, N-Nus, N-EK |
載體抗性: | Ampicillin |
表達宿主菌: | BL21(DE3) 、 Rosetta2(DE3)、C43(DE3) |
備注: |
C term His tag only; N term thrombin cleavage site; N term enterokinase cleavage site; a,b,c vary by MCS
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產品目錄號: | 70939-3 |
穩定性: | 瞬時表達 Transient |
組成型: | 組成型 Constitutive |
病毒/非病毒: | 非病毒 |
Feature Name | Start | End | |
---|---|---|---|
T7_Terminal_primer | 68 | 86 | |
rrnB_T2_terminator | 167 | 140 | |
rrnB_T1_terminator | 342 | 299 | |
rrnB_terminator | 465 | 308 | |
EK | 694 | 680 | |
S15 | 790 | 746 | |
6xHIS | 817 | 800 | |
T7_transl_en_RBS | 2342 | 2326 | |
lacO | 2387 | 2360 | |
T7_promoter | 2405 | 2387 | |
tet (300 - 563) | 2441 | 2704 | |
pBRrevBam_primer | 2512 | 2493 | |
lacI | 2787 | 3878 | |
ROP | 4450 | 4641 | |
pGEX_3_primer | 4657 | 4635 | |
pBR322_origin | 5675 | 5056 | |
AmpR_promoter | 5762 | 5790 | |
Ampicillin | 5833 | 6693 |
ORF | Start | End | |
---|---|---|---|
ORF frame 3 | 2317 | 548 | |
ORF frame 2 | 617 | 2131 | |
ORF frame 2 | 2318 | 3046 | |
ORF frame 3 | 2919 | 3878 | |
ORF frame 1 | 5833 | 6693 |
Enzyme Name | Cut |
---|---|
PacI | 502 |
XhoI | 529 |
PmlI | 575 |
NotI | 591 |
EagI | 591 |
HindIII | 598 |
KpnI | 623 |
SalI | 625 |
PstI | 638 |
AscI | 640 |
EcoRI | 653 |
BamHI | 659 |
SacI | 671 |
SmaI | 720 |
XmaI | 718 |
SacII | 796 |
SpeI | 827 |
BglII | 942 |
MscI | 1074 |
StuI | 1127 |
NcoI | 1402 |
NdeI | 2316 |
XbaI | 2354 |
ApaI | 3357 |
HpaI | 3652 |
FspI | 6397 |
The pET-43.1 series of vectors are designed for cloning and high-level expression of peptide sequences fused with the 491 aa Nus?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 ColiDOWN primer (cat. no. 70845-3). Vector encoded sequence can be completely removed when cloning into the PshA I or Sma I sites (as shown below) by cleaving the Nus?Tag fusion protein with enterokinase or thrombin, respectively.