pET-42b(+)載體基本信息
| 別名: | pET42b, pet 42b, pET-42b(+) |
|---|---|
| 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: | 5929bp |
| 5' 測序引物: | T7 |
| 5' 測序引物序列: | 5'-TAATACGACTCACTATAGGG-3' |
| 載體標簽: | N-GST, N-His, C-His |
| 載體抗性: | Kanamycin |
| 備注: |
含有GST融合蛋白;N端Thrombin蛋白酶切位點; 含有N端Factor Xa蛋白酶切位點 ; pET 42 a,b,c 的差異僅在多克隆位點區域。
|
| 產品目錄號: | 70562-3 |
| 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_terminator | 120 | 1 | |
| T7_Terminal_primer | 69 | 87 | |
| S15 | 350 | 306 | |
| 6xHIS | 410 | 393 | |
| GST (variant) | 1091 | 441 | |
| T7_transl_en_RBS | 1116 | 1100 | |
| lacO | 1161 | 1134 | |
| T7_promoter | 1179 | 1161 | |
| tet (300 - 563) | 1215 | 1478 | |
| pBRrevBam_primer | 1286 | 1267 | |
| lacI | 1561 | 2652 | |
| ROP | 3224 | 3415 | |
| pGEX_3_primer | 3431 | 3409 | |
| pBR322_origin | 4449 | 3830 | |
| KanR2 | 4555 | 5370 |
| ORF | Start | End | |
|---|---|---|---|
| ORF frame 3 | 1091 | 147 | |
| ORF frame 3 | 1092 | 1820 | |
| ORF frame 1 | 1693 | 2652 | |
| ORF frame 1 | 4555 | 5370 |
| 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 | 288 |
| KpnI | 295 |
| BglII | 298 |
| SacII | 389 |
| SpeI | 420 |
| MscI | 883 |
| NdeI | 1090 |
| XbaI | 1128 |
| ApaI | 2131 |
| HpaI | 2426 |
| NruI | 4643 |
| SmaI | 4860 |
| XmaI | 4858 |
The pET-42 series is designed for cloning and high-level expression of peptide sequencesfused with the 220 aa GST?Tag? protein. Unique sites are shown on the circle map. Note that thesequence is numbered by the pBR322 convention, so the T7 expression region is reversed on thecircle map. The cloning/expression region of the coding strand transcribed by T7 RNA polymeraseis shown below. The f1 origin is oriented so that infection with helper phage will produce virionscontaining single stranded DNA that corresponds to the coding strand. Therefore, single strandedsequencing should be performed using the T7 terminator primer (cat. no. 69337-3). Vector encodedsequence can be completely removed when cloning into the PshAI site (as shown below) and thencleaving the GST fusion protein with Factor Xa.