Plasmid #34 enables target proteins to be labeled with C-terminal His8 and CL7. Protease cleavage removes both tags.
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 Sortase A (SRT) and PreScission protease (PSC) cleavage sites and a His8 tag.
CLEAVAGE SITE(S)
N-terminal SUMO and C-terminal SRT and PSC cleavage sites are available in Plasmid #34.
OTHER TAGS
The encoded region begins with a Trx tag and includes a C-terminal His8 tag upstream of CL7.
CLONING OPTIONS
1. HindIII/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | PSC | PSC | PSC | His8 | 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 | PSC | PSC | PSC | His8 | CL7
This insertion option results in 2 extra aa 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 | His8 | 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:
