Part:BBa_K2557051

EF1α promoter-TetR

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. In order to increase TetR expression to enhance repression, we combined TetR and EF1α promoter.

Characterization

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, However, we did not analyze the expression level of TetR initiated by the EF1α promoter.

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 581