Part:BBa_K3132100

UAS_MinimalCMV Promotor

This part (4*UAS MinimalCMV) is one of our synthetic promoters (SynPro) based on minimalCMV promoter (BBa_K3132004) and 4*GAL4 binding sites (BBa_K3132014). We inset the 4*Gal4 binding sites before the minimalCMV promoter. MinimalCMV promoter is a promoter from the vector pcDNA3.1 with very strong transcriptional activity. In this way, we can use our synthetic transcription factors (SynTFs) which is a fusing protein Gal4_DBD_(G4S) linker_NLS_VP64. The Gal4-DBD can bind to the 4*Gal4 binding sites specifically[1] and the VP64 domain can initiate transcription by activating the 4*UAS MinimalCMV promoter. The corresponding SynTF of UAS_minimalCMV is Gal4-VP64 (BBa_K3132000).

Characterization of UAS_MinimalCMV Promotor:

We used fluorescence protein mCherry to verify the binding of Gal4-DBD to 4*UAS(Gal4 binding sites), and the results are as follows. We inset mCherry behind 4*UAS and in general the fluorescence intensity is very low; when Gal4 is added, the fluorescence intensity significantly increased which verifies the binding of Gal4-DBD to 4*UAS.

References

[1] L. Morsut et al., Engineering Customized Cell Sensing and Response Behaviors Using Synthetic Notch Receptors. Cell 164, 780--791 (2016).

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
  COMPATIBLE WITH RFC[1000]