Fig. 2

Cloning and expression region of pCold-II-TEV. The cold-shock protein A promoter (cspA), the lac operator, ribosome binding site (RBS) and the translation-enhancing element (TEE) regulate gene expression from pCold-based vectors. The cspA-derived 5′UTR region allows low-temperature induction as it is stable at temperatures around 15 °C, but highly unstable at 37 °C. A TEV protease recognition sequence was introduced at the BamHI restriction site of the cloning region of pCold-II to allow removal of the vector-encoded His-tag from recombinant proteins. The cartoon is drawn to scale