plasmid 32 option 1
plasmid #32 option 2
plasmid #32 option 3
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Plasmid #32

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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.

    1. 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.

    1. Bsu36I/SpeI Insertion Site – Trx | SUMO | Gene of Interest | SRT | 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 #32