Regulatory

Part:BBa_K1723001

Designed by: Emilie Cuillery   Group: iGEM15_EPF_Lausanne   (2015-09-08)
Revision as of 12:32, 17 September 2015 by E.Cuillery (Talk | contribs)

PAM rich URS j23117 promoter


PAM rich URS J23117 is an improvement of the J23117 (BBa_J23117) weak promoter [1]. This promoter is the promoter J23117 flanked with a PAM (PAM = NGG sequence) rich Upstream Regulatory Sequence (URS) to enable the use of protein dCas9-ω (BBa_K1723000) as a gene transcription regulator when in complex with one sgRNA targeting the promoter such as: the activator sgRNA Z0 (BBa_K1723002), the inhibitor sgRNA Z35 (BBa_K1723003), or the other inhibitor sgRNA Z4 (BBa_K1723004). dCas9 can only bind if its target sequence is preceded PAM sequence. Discover the applications of this part in the tab 'experience'.

Discover all the parts that can work with this one:

http://2015.igem.org/Team:EPF_Lausanne/Part_Collection

EPFL_Lausanne_promoter.png

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 56
    Illegal NheI site found at 79
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 43
  • 1000
    COMPATIBLE WITH RFC[1000]

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.


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