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pASK-IBA6C is a classic cloning vector developed for the E. coli expression of recombinant proteins with Strep-tag®II fused to the N-terminus. The vector carries the chloramphenicol resistance cassette for selection of transformed E. coli cells, F1 and ColE1 origin for a high plasmid copy number, inducible tetracycline promoter/operator for the regulated expression of proteins, multiple cloning site with recognition sites for several common restriction enzymes (EcoRI, BamHI, SmaI, SalI, PstI), and the ompA signal for periplasmic secretion of the recombinant protein.
The vector can be combined with any E. coli strain since the tet-promoter works independently of the genetic background of E. coli. Expression of the recombinant protein is induced by the addition of anhydrotetracycline, which is a derivative of tetracycline and does not exhibit antibiotic activity. During secretion into the periplasmic space, the ompA signal is cleaved off, and the N-terminal Strep-tag®II can be removed by protease factor Xa.
