Generator

Part:BBa_K1486008:Design

Designed by: iGem EPFL 2014   Group: iGEM14_EPF_Lausanne   (2014-08-26)
Revision as of 12:22, 21 September 2014 by Robert Baldwin (Talk | contribs) (→‎References)


CxpR & Split IFP1.4 [Cterm + Cterm][1]


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 2213
    Illegal PstI site found at 3446
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
    Illegal PstI site found at 2213
    Illegal PstI site found at 3446
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 3657
    Illegal BamHI site found at 1144
    Illegal XhoI site found at 1283
    Illegal XhoI site found at 2470
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 2213
    Illegal PstI site found at 3446
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 2213
    Illegal PstI site found at 3446
    Illegal AgeI site found at 979
    Illegal AgeI site found at 1694
    Illegal AgeI site found at 2881
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

This construct aimed to evaluate the activation and presumed dimerization of CpxR in E.Coli by fusing split IFP1.4 (Infrared Fluorescent Protein). When dimerizing, CpxR allows the two parts of the split protein to re-fold and acquire their ability to emit fluorescence. Not knowing how CpxR might dimerize, we built 4 different biobricks with the various possible orientations that the dimerization of CpxR might acquire.

It is to be noted that this construct makes use of an Arabinose-inducible promoter, a standard Elowitz RBS, as well as a flexible linker 2 x (Gly-Gly-Gly-Gly-Ser) between CpxR and the two split parts of the IFP

Source

http://www.addgene.org/browse/article/8177/

References

Michnick, S., Tchekanda, E., & Sivanesan, D. (2014, April 20). An infrared reporter to detect spatiotemporal dynamics of protein-protein interactions. Nature Methods, 6-6.