Plasmid #9 allows for the expression of membrane proteins with C-terminal His8 and CL7 tags. It also encodes an engineered signal peptide from the Bacillus halodurans YidC gene. Membrane proteins are inserted at the BamHI/SpeI restriction sites where there is an existing 754 base pair restriction fragment.
Transcription is induced with IPTG and driven by the T7 RNA polymerase. The plasmid is designed for expression in E. coli.
The CL7 tag for purification is located at the C-terminus, downstream of a PreScission protease (PSC) cleavage site and His8 tag.
Plasmid #9 includes a PSC cleavage site directly adjacent to the target protein and upstream of both the His8 and CL7 purification tags.
The plasmid contains a His8 tag located downstream of the target protein and cleavage tag but upstream of the CL7 tag.