Coding

Part:BBa_K3132103

Designed by: Qiliang Yang   Group: iGEM19_SMMU-China   (2019-10-14)


ZF43-8_MinimalCMV Promotor

This part (8* ZF_43-8 MinimalCMV) is one of our synthetic promoters (SynPro) based on minimalCMV promoter (BBa_K3132004) and eight repeats of Zinc Finger_43-8 binding sites. We inset the 8* ZF_43-8 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 ZF_43-8 _DBD_(G4S) linker_NLS_VP64. The ZF_43-8-DBD can bind to the 8* ZF_43-8 binding sites specifically[1] and the VP64 domain can initiate transcription by activating the MinimalCMV 8*PIP promoter. The corresponding SynTF of ZF_43-8_minimalCMV is ZF43-8-VP64 (BBa_K3132001).

Characterization of ZF43-8_MinimalCMV Promotor:

We used fluorescence protein mCherry to verify the binding of ZF_43-8-DBD to 8* ZF_43-8(Zinc Finger_43-8 binding sites), and the results are as follows. We inset mCherry behind 8* ZF_43-8 and in general the fluorescence intensity is very low; when ZF_43-8 is added, the fluorescence intensity significantly increased which verifies the binding of ZF_43-8-DBD to 8* ZF_43-8.

T--SMMU-China--UAS MinimalCMV Promotor.png

References

[1] J. J. Lohmueller, T. Z. Armel, P. A. Silver, A tunable zinc finger-based framework for Boolean logic computation in mammalian cells. Nucleic Acids Research 40, 5180--5187 (2012).

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]


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Categories
Parameters
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