plasmid 33 option 1
plasmid #33 option 2
plasmid #33 option 3
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Plasmid #33

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Plasmid #33 enables target proteins to be labeled with C-terminal His8 and CL7. The His8 tag remains attached depending on the protease utilized.

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) cleavage sites, one Sortase A (SRT) cleavage site, and a His8 tag.
Cleavage Site(s)

Plasmid #33 includes an N-terminal SUMO cleavage site, three PSC, and one SRT C-terminal cleavage sites.
Other Tags

The encoded region begins with a Trx tag. A C-terminal His8 tag is also present upstream of the CL7 tag.
Cloning Options


    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.

    1. KpnI/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | His8 | PSC | PSC | PSC | CL7


This insertion option results in 2 extra aa at the N terminus of the target following SUMO protease cleavage.

    1. 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:

Plasmid #33