质粒类型: | 大肠杆菌表达载体 |
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表达水平: | 高 |
克隆方法: | 多克隆位点,限制性内切酶 |
载体大小: | 5917 bp (查看载体序列) |
5' 测序引物: | T7或者Trx-F |
5' 测序引物序列: | T7: 5'-TAATACGACTCACTATAGGG-3'; Trx-F: 5' TTCCTCGACGCTAACCTG 3' |
载体标签: | thioredoxin (N端); His (中间和C端), N-EK |
载体抗性: | Ampicillin (氨苄青霉素) |
备注: |
For directional cloning of PCR-amplified DNA; ligation independent cloning; enterokinase cleavage site.
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The pET-32 Ek/LIC vector is prepared for rapid, directional cloning of PCR-amplified DNA for high-level expression of polypeptides fused with the 109aa Trx•Tag™ thioredoxin protein. Using specifically designed primers for amplification and the pET-32 Ek/LIC Cloning Kit (Cat. No. 69076-3), inserts can be efficiently cloned without the need for restriction digestion or ligation. Fusion proteins also contain 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 (Cat. No. 69337-3).