pET-45b(+)載體基本信息
別名: | pET45b, pET 45b |
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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: | 5260 bp |
5' 測序引物: | T7 |
5' 測序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3' |
3' 測序引物: | T7t |
3' 測序引物序列: | T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
載體標簽: | N-His,N-EK, C-S |
載體抗性: | Ampicillin |
表達宿主菌: | BL21(DE3) 、 Rosetta2(DE3)、C43(DE3) |
備注: |
N-term HisTag with minimal extraneous sequences.
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產品目錄號: | 71327-3 |
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 | |
---|---|---|---|
Ampicillin | 1442 | 582 | |
AmpR_promoter | 1513 | 1485 | |
pBR322_origin | 1600 | 2219 | |
pGEX_3_primer | 2618 | 2640 | |
ROP | 2825 | 2634 | |
lacI | 4488 | 3397 | |
tet (887 - 636) | 4831 | 4580 | |
pBRrevBam_primer | 4763 | 4782 | |
T7_promoter | 4870 | 4888 | |
lacO | 4888 | 4915 | |
T7_transl_en_RBS | 4933 | 4949 | |
EK | 4999 | 5013 | |
T7_Terminal_primer | 5192 | 5174 | |
T7_terminator | 5193 | 5228 |
ORF | Start | End | |
---|---|---|---|
ORF frame 3 | 1442 | 582 | |
ORF frame 2 | 4356 | 3397 | |
ORF frame 1 | 4729 | 4229 |
Enzyme Name | Cut |
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FspI | 877 |
HpaI | 3622 |
ApaI | 3921 |
BclI | 4110 |
ClaI | 4849 |
XbaI | 4916 |
NcoI | 4955 |
PmlI | 4980 |
KpnI | 4988 |
AgeI | 4987 |
BstBI | 4993 |
BamHI | 5019 |
SacI | 5036 |
AscI | 5047 |
PstI | 5057 |
SalI | 5059 |
HindIII | 5065 |
NotI | 5072 |
EagI | 5072 |
XhoI | 5080 |
PacI | 5155 |
The pET-45b(+) vector carries an N-terminal His?Tag® coding sequence followed by an enterokinase cleavage site and an optional C-terminal S?TagTM 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 vector map. 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).