Plasmid #21

Plasmid #21 enables the tagging of target proteins with N-terminal CL7 and His8. There are three options for the 5’ placement of target genes (see cloning options below). XhoI is the 3’ cloning site.  All three options require a stop codon encoded in the 3’ cloning primer. 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 N-terminal CL7 tag is upstream of several protease cleavage sites and a His8 tag. CLEAVAGE SITE(S): 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. OTHER TAGS: 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.CLONING OPTIONS: 1. HindIII/XhoI Insertion Site – Trx | CL7 | PSC | 44-Amino Acid Linker | PSC | His8 Tag | SUMO | 13-Amino Acid Linker | SRT | Gene of Interest 2. KpnI/XhoI Insertion Site – Trx | CL7 | PSC | 44-Amino Acid Linker | PSC | His8 Tag | SUMO | 3-Amino Acid Linker | Gene of Interest 3. Bsu36I/XhoI Insertion Site – Trx | CL7 | PSC | 44-Amino Acid Linker | PSC | His8 Tag | SUMO | Gene of Interest EXPIRATION:  6 months from receipt when stored as directed. Plasmid guide with plasmid sequence file is available at Trialtusbioscience.com.