Plasmid #33 enables target proteins to be labeled with C-terminal His8 and CL7. The His8 tag remains attached depending on the protease utilized.
Transcription is induced with IPTG and driven by the T7 RNA polymerase. The plasmid is designed for expression in E. coli.
The C-terminal CL7 tag is downstream of three PreScission protease (PSC) cleavage sites, one Sortase A (SRT) cleavage site, and a His8 tag.
Plasmid #33 includes an N-terminal SUMO cleavage site, three PSC, and one SRT C-terminal cleavage sites.
The encoded region begins with a Trx tag. A C-terminal His8 tag is also present upstream of the CL7 tag.
1. HindIII/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | His8 | PSC | PSC | PSC | CL7
This insertion option results in 7 extra aa at the N terminus of the target following SUMO protease cleavage.
2. KpnI/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | His8 | PSC | PSC | PSC | CL7
3. Bsu36I/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | His8 | PSC | PSC | PSC | CL7
This insertion option maintains the Gene of Interest’s wildtype sequence, without adding any extra residues. The N-terminus of the Gene of Interest must include the following to complete the SUMO C-terminal sequence:
EXPIRATION | 6 months from receipt when stored as directed
Plasmid Sequences are available on the Plasmid Guide Page
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