别名: | pET49b, pET 49b |
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质粒类型: | 大肠杆菌表达载体 |
表达水平: | 高 |
克隆方法: | 多克隆位点,限制性内切酶 |
载体大小: | 5926 bp (查看载体序列) |
5' 测序引物: | T7或者pGEX 5' |
5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3'; pGEX 5': 5'-GGGCTGGCAAGCCACGTTTGGTG-3' |
3' 测序引物: | T7t |
3' 测序引物序列: | T7t: 5'-TGCTAGTTATTGCTCAGCGG-3' |
载体标签: | N-GST, N-His, N-HRV 3C, C-S, C-Thrombin |
载体抗性: | Kanamycin (卡那霉素) |
备注: |
Same as pET47 but also has Nterm GST Tag; contains HRV 3C Protease cleavage site for fusion tag removal at low temperatures; Cterm thrombin cleavage site.
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The pET-49b(+) vector carries N-terminal GST•Tag™ and His•Tag? coding sequences followed by a recognition site for the human rhinovirus (HRV) 3C protease. This protease is highly specifi c for cleavage of the sequence LEVLFQ↓GP , and is active at low temperatures . pET-49b(+) also contains an optional C-terminal thrombin recognition site followed by an S•Tag™ coding sequence. Unique restriction .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 the helper phage will produce virions containing single-stranded DNA that corresponds to the coding strand. Therefore, single-stranded sequencing should be performed using the AS S•Tag 18mer Primer (Cat. No. 71262-3).