Plasmid #21 enables target proteins to be tagged with N-terminal CL7 and His8 and includes the HIV Nef protein as a model (inserted with HindIII/XhoI restriction sites).
Transcription is driven by the T7 RNA polymerase, and the plasmid is designed for expression in E. Coli.
The N-terminus CL7 tag is upstream of several protease cleavage sites and a His8 tag.
Plasmid #21 contains two PreScission protease (PSC) cleavage sites between the CL7 and His8 tags. Downstream of the His8 tag are SUMO and Sortase A cleavage sites.
The coding region begins with an N-terminal Trx tag and includes a His8 tag between the PSC and SUMO cleavage sites, upstream of the target protein.