Part:BBa_K4190016

TEV Cut Site

Cut site for eventual removal of His-tag.

Usage and Biology

For the gene insert containing the sequence to produce the GLP-1 receptor in E. coli, the enterokinase cut site was replaced with a TEV protease cut site. This was done because our lab was given TEV protease by the Dubois Lab at UCSC.

A codon optimized sequence for the GLP1 receptor was also added to a separate gene insert for binding affinity testing. Since this gene insert contained a TEV protease cut site rather than an enterokinase cut site, an alanine was coded for in between the cut site and the GLP1 receptor sequence. This is because TEV protease will only cut before serine, glycine, alanine, methionine, cysteine, or histidine [1].

Sequences and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

References

[1] “TEV Protease | NEB.” https://www.neb.com/products/p8112-tev-protease (accessed Aug. 27, 2022).