Regulatory

Part:BBa_K2398003:Design

Designed by: Moritz Przybilla   Group: iGEM17_Heidelberg   (2017-10-25)


Constitutive Promotor with RFP guide RNA for application in Phage-assisted continous evolution (PACE


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 159
    Illegal NheI site found at 182
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1
    Illegal BglII site found at 223
    Illegal XhoI site found at 72
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 146
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

This part is based in a plasmid from Qi et al. [3]
This was designed for the use with the cloning standard of the iGEM Team Heidelberg 2017 (http://2017.igem.org/Team:Heidelberg/RFC). It is meant for the usage as building block for accessory plasmids in the context of phage assisted continuous ecolution [1] or related systems [2].

The part is fully compatible with RFC10, as well as with Golden Gate assembly


Source

PCR amplified from plasmid

References

[1] Esvelt, Kevin M.; Carlson, Jacob C.; Liu, David R. (2011): A system for the continuous directed evolution of biomolecules. In: Nature 472 (7344), S. 499–503. DOI: 10.1038/nature09929.

[2]Brödel, Andreas K.; Jaramillo, Alfonso; Isalan, Mark (2016): Engineering orthogonal dual transcription factors for multi-input synthetic promoters. In: Nature communications 7, S. 13858. DOI: 10.1038/ncomms13858. 

[3] Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific Control of Gene Expression. Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim WA. Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022. 10.1016/j.cell.2013.02.022 PubMed 23452860