pET-35b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： 5927bp

质粒标签： N-CBDclos,C-6×His

原核抗性： 卡那霉素Kan

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

质粒简介

The pET-35b(+) vector is designed for expression of CBDclos fusion proteins.A variety of cloning sites and strategies are available. Rapid directional cloning of PCR-amplified DNA is available with the pET-35 Xa/LIC Vector Kit, which contains linearized pET-35b(+) ready for annealing with appropriately prepared inserts. When cloned with the Xa/LIC method, resulting CBDclos•Tag™ fusion proteins can be cleaved precisely at the vector-encoded junction using Factor Xa. Unique sites are shown on the circle map. Note that the sequence is num-bered 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.