ligation independent cloning (LIC) clone with pBR322 origin and Ampicillin Resistance and ROP and ORF frame 2 – Tetracycline resistance and ORF frame 1 and pBRrevBam and T7 and T7 term primer and BseRI Stuffer and MBP signal sequence (M1-A26 of NP_290668) and RBS and MBP (K27-T392 of NP_290668) and HIS10 and TEV and FC sequencing primer pET26 pelB seq R and FC sequencing primer MBP pelB seqF 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 T7 term and FC sequencing primer pET-PPL seq 2 R and AmpR; ampicillin 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