Part:BBa_K3132101

PIP_MinimalCMV Promotor

This part (8*PIP MinimalCMV ) is one of our synthetic promoters (SynPro) based on minimalCMV promoter  (BBa_K3132004)  and eight repeats of PIP binding sites. We inset the 8*PIP 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 PIP_DBD_(G4S) linker_NLS_VP64. The PIP-DBD can bind to the 8*PIP binding sites specifically[1] and the VP64 domain can initiate transcription by activating the MinimalCMV 8*PIP promoter. The corresponding SynTF of PIP_minimalCMV is PIP-VP64 (BBa_K3132003).

Characterization of PIP_MinimalCMV Promotor:

We used fluorescence protein mCherry to verify the binding of PIP-DBD to 8*PIR(PIP binding sites), and the results are as follows. We inset mCherry behind 8*PIR and in general the fluorescence intensity is very low; when PIP is added, the fluorescence intensity significantly increased which verifies the binding of PIP-DBD to 8*PIR.

References

[1] M. Fussenegger et al., Streptogramin-based gene regulation systems for mammalian cells. Nature biotechnology 18, 1203--1208 (2000).

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]