ligation independent cloning (LIC) clone, bacterial expression expression with RBS (extended) and TET-2 and pelB and pBR322 origin and KanR and ROP and BseRI and HIS10 and MBP and TEV and FC sequencing primer pET26 pelB seq R and FC sequencing primer MBP pelB seqF and TET and LacI and FC sequencing primer pET-PPL seq 1 F and FC sequencing primer pET-PPL seq 1 R and FC sequencing primer pET-PPL seq 2 F and pBRrevBam and T7 and lac operator and T7 term primer; kanamycin resistance in bacterial;
Comments:
TEV protease cleavage site is designed for use with ligation-independent cloning into the BseRI site. This TEV site contains only the amino acids DNLYFQ. Cleavage occurs after the Q, and requires a compatible amino acid C-terminal to the Q, such as S, G, A, M. See Kapust et al 2002 Biochem Biophys Res Commun 294:949