Regulatory

Part:BBa_K1723025:Design

Designed by: Cyril Pulver   Group: iGEM15_EPF_Lausanne   (2015-09-18)


CYC_3 promoter


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 13
    Illegal XbaI site found at 197
    Illegal PstI site found at 19
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 13
    Illegal PstI site found at 19
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 13
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 13
    Illegal XbaI site found at 197
    Illegal PstI site found at 19
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 13
    Illegal XbaI site found at 197
    Illegal PstI site found at 19
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This promoter was specifically designed to be controlled by complexes constituted of dCas9_VP64 and gRNAs c3_1, c6_1 and c7_1. The design of the promoter is based on promoter CYC_0 (BBa_K1723022). It shows specific mutations that make c3_3, c6_3 and c7_3 sites differ from c3_0, c6_0 and c7_0 sites which are present on CYC_0. Since it has been demonstrated with CYC_0 that inhibition prevails over activation when combining the co-expression of dCas9_VP64 and those gRNAs [1], this promoter can be used as a basic cellular computation tool.

Mutations were carefully designed to keep both TATA sequences [2] of CYC_0 intact.


Source

Synthesized as a G-Block


References

[1] Farzadfard, F., Perli, S.D., Lu, T.K., 2013. Tunable and Multifunctional Eukaryotic Transcription Factors Based on CRISPR/Cas, ACS Synthetic Biology (ACS publications), http://pubs.acs.org/doi/pdf/10.1021/sb400081r (16.09.2015)

[2] Hahn, S., Buratowski, S., Sharp, P.A., Guarente, L, 1989. Yeast TATA-binding protein TFIID binds to TATA elements with both consensus and nonconsensus DNA sequences, Proc. Natl. Acad. Sci. U.S.A. Pubmed, https://www.wikigenes.org/e/ref/e/2569738.html (18.09.2015)