Generator

Part:BBa_K1486043:Design

Designed by: EPFL iGem team 2014   Group: iGEM14_EPF_Lausanne   (2014-10-02)


Leucine Zipper + split rLuc


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 961


Design Notes

When designing the part, we took care of adding a start codon at the beginning of the C terminus part of the split luciferase and deleting the stop codon at the end of the same sequence.

The two fusion proteins for the experiment mentionned in the main page are on the same plasmid. They are separated by a short random sequence followed by a second RBS.

We took care of deleting the start codon at the beginning of the zippers too.

Source

Zipper sequences were obtained from here : http://www.addgene.org/browse/article/8177/

Split renilla luciferase sequences were obtained by PCR amplification of the plasmid prLuc from Waldor Laboratory [http://waldorlab.bwh.harvard.edu/] used in this paper [http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0043175].

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.