The pET-32a-c series is designed for cloning and high-level expression of peptide sequences fused with the 109aa Trx?Tag? thioredoxin protein (1). Cloning sites are available for producing fusion proteins also containing cleavable His?Tag? and S?Tag? sequences for detection and purification. 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 T7 terminator primer.
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 | 5899 bp |
5'测序引物及序列 | T7:5'-TAATACGACTCACTATAGGG-3' |
3'测序引物及序列 | T7t:5'-GCTAGTTATTGCTCAGCGG-3' |
载体标签 | N-T7,C-His |
载体抗性 | 氨苄青霉素(Ampicillin) |
备注 | Production of soluble, active target proteins; N-term thrombin cleavage site; Nterm enterokinase cleavage site; a,b,c vary by MCS |
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