pET-31b质粒

基本信息

启动子： T7/lac

复制子： ColE1 ori，F1 ori

终止子： T7 terminator

质粒分类： 大肠杆菌载体；PET系列表达质粒

质粒大小： 5742bp

质粒标签： C-6×His

原核抗性： 氨苄青霉素Amp

克隆菌株： DH5α

培养条件： 37℃，有氧，LB

表达宿主： BL21(DE3)

诱导方式： IPTG或乳糖及其类似物

5'测序引物： T7：TAATACGACTCACTATAGGG

3'测序引物： T7-ter：TGCTAGTTATTGCTCAGCGG

备注： 表达酮固醇异构酶

质粒简介

The pET-31b(+) vector is designed for cloning and high-level expression of peptide sequences fused with the 125aa ketosteroid isomerase protein (1). The unique AlwN I cloning site allows the unidirectional insertion of of tandemly repeated peptide coding regions separated by methionine codons. 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 circular 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 .