Plasmid #32

Plasmid #32 enables labeling of target proteins with an uncleavable N-terminal His8 and a cleavable C-terminal CL7 tag. EXPRESSION: Transcription is induced with IPTG and driven by the T7 RNA polymerase. The plasmid is designed for expression in E. coli. PEPTIDE TAG: The C-terminal CL7 tag is downstream of three PreScission protease (PSC) and one Sortase A (SRT) cleavage site. CLEAVAGE SITE(S): An N-terminal SUMO cleavage site exists upstream of the His8 tag. One SRT and three PSC cleavage sites exist between the target protein and the CL7 tag. OTHER TAGS: Plasmid #32 includes N-terminal Trx and His8 tags. CLONING OPTIONS: 1. HindIII/SpeI Insertion Site – Trx | SUMO | His8 | Gene of Interest | SRT | PSC | PSC | PSC | CL7 This insertion option results in 16 extra aa (including His8 tag) at the N terminus of the target following SUMO protease cleavage. 2. KpnI/SpeI Insertion Site – Trx | SUMO | His8 | Gene of Interest | SRT | PSC | PSC | PSC | CL7 This insertion option results in 11 extra aa (including His8 tag) at the N terminus of the target following SUMO protease cleavage. 3. Bsu36I/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | PSC | PSC | PSC | CL7 EXPIRATION: 6 months from receipt when stored as directed. Plasmid guide with plasmid sequence file is available at Trialtusbioscience.com.