I am currently using a lac-based vector for periplasmic expression of proteins in E. coli. I now want to re-design the vector and express the proteins under the control of the T7 promoter in a DE3 strain. I found a lot of different vector sequences and am a bit confused which sequence parts I really need in my vector for IPTG- inducible expression of my proteins.
T7 promoter only? T7 promoter and T7 terminator? T7 promoter plus lac operator? (If I got that right, the T7 RNAP is under the control of a lac promoter in the bactrial genome, so I shouldn't need a lac operator in my vector?) Which ribosome binding motif?
I am looking forward to your replies!