Part:BBa_K2557050

TetR with TEV cleavage site

TetR is a repressor that binds to TetO sequences and inhibits the expression of downstream genes. iGEM17_Oxford inserted a TEV cleavage site in TetR (BBa_K2450202), so TetR can be recognized and cleaved by TEV protease, losing binding function and releasing inhibition of downstream genes. In our system, we use this TetR as a negative regulatory protein of the recombinase.

Usage and Biology

Fig.1 Inhibition of tetR on promoter with tetO sequence (A)Fluorescence microscope observation of HEK 293T only transfect with plasmids containing promoters with tetO sequence. (B)Fluorescence microscope observation of HEK 293T transfect with plasmids containing promoters with tetO sequence and tetR.

Fig. 1 shows that TetR has obvious inhibitory effects on the three promoters downstream of TetO.

Improvement

This part was modified from BBa_K2450202, which we codon optimized and characterized in HEK 293T.

References

1. Ramos, J. L. et al. The TetR Family of Transcriptional Repressors The TetR Family of Transcriptional Repressors. Microbiol. Mol. Biol. Rev. 69, 326–356 (2005).

Sequence 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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 361