2、28b载体质粒图谱和多克隆位点信息pET28b载体简介pET‐28b载体带有一个N端的His/Thrombin/T7蛋白标签，同时含有一个可以选择的C端His标签。pET28b载体的单一的多克隆位点见上面的环状质粒图谱。注意：载体序列是以pBR322质粒的编码规矩进行编码的，所以T7蛋白表达区在质粒图谱上面是反向的。T7 RNA聚合酶启动的克隆和表达区域在质粒图谱中也被标注了出来。质粒的F1复制子是被定向的，所以在T7噬菌体聚合酶的作用下，包含有蛋白编码序列的病毒粒子能够产生，并启动蛋白表达，同时蛋白表达将被T7终止子序列(Cat. No. 69337‐3)的作用下终止蛋白翻译。The p
3、ET‐28a‐c(+) vectors carry an N‐terminal His•Tag®/ thrombin/ T7•Tag® configuration plus an optional C‐terminal His•Tag sequence. 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 cl
4、oning/expression region of the coding strand transcribed by T7 RNA polymeraseis 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 perf
5、ormed usingthe T7 terminator primer (Cat. No. 69337‐3).pET28b载体序列ORIGIN1ATCCGGATATAGTTCCTCCTTTCAGCAAAAAACCCCTCAAGACCCGTTTAGAGGCCCCAA61GGGGTTATGCTAGTTATTGCTCAGCGGTGGCAGCAGCCAACTCAGCTTCCTTTCGGGCTT121TGTTAGCAGCCGGATCTCAGTGGTGGTGGTGGTGGTGCTCGAGTGCGGCCGCAAGCTTGT181CGACGGAGCTCGAATTCGGATCCCGACCCATTTGCTGT
