Regulatory

Part:BBa_K1723005:Design

Designed by: Emilie Cuillery   Group: iGEM15_EPF_Lausanne   (2015-09-15)
Revision as of 21:58, 16 September 2015 by E.Cuillery (Talk | contribs) (→‎Design Notes)


PAM rich URS J23117Alt promoter


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 277
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 167
    Illegal XhoI site found at 195
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part was synthesized on the model of the PAM rich URS J23117 promoter (BBa_K1723001). It was obtained by mutating bases on the promoter to obtain different target sites for sgRNAs for promoter regulation using dCas9-ω system. We mutated the sequences as we wanted except for the -35 sequence and -10 sequence and -35 sequence to keep a similar promoter strength [1].

Source

this part was fully synthesized.

References

[1] Bikard, D., Jiang, W., Samai, P., Hochschild, A., Zhang, F., & Marraffini, L. A. (2013). Programmable repression and activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic acids research, 41(15), 7429-7437.

[2] Qi, L. S., Larson, M. H., Gilbert, L. A., Doudna, J. A., Weissman, J. S., Arkin, A. P., & Lim, W. A. (2013). Repurposing CRISPR as an RNA-guided platform for sequence-specific control of gene expression. Cell, 152(5), 1173-1183.

[3] Alec AK Nielsen & Christopher A Voigt (2014). Multi-input CRISPR/Cas circuits that interface host regulatory network. Molecular systems biology, 10(11), 763.