The plasmid pSpot3 is an expression vector for Spot-Tag® fusion proteins in E. coli. After cloning the protein of interest (POI) into the multiple cloning site (MCS) provided by pSpot3, the Spot-Tag® (sequence: PDRVRAVSHWSS) will be fused to the C-terminus of the POI.
Protein expression can be induced using lactose or IPTG, but requires the use of DE3 prophage-positive E. coli expression strains. The resulting Spot-Tag® fusion protein can be purified using the ChromoTek Spot-Trap®.
Suitable host strains for propagation in E. coli include DH5alpha, HB101, XL1-Blue, and other general purpose strains. The vector confers resistance to ampicillin (100 μg/ml) to E. coli hosts. (This plasmid was designed and generated by Dr. Philipp Kaiser from the NMI.)