Coding

Part:BBa_K2483007:Design

Designed by: Bryan Nowack   Group: iGEM17_Potsdam   (2017-09-17)


Ddx4-YFP


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 790
    Illegal BamHI site found at 802
    Illegal BamHI site found at 1531
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 978
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 173
    Illegal SapI.rc site found at 832


Design Notes

We codon optimized it for yeast and changed the linker to a 10x SG linker. This is because of our following experiments (BBa_2483010) where we had bigger enzymes than YFP and wanted to make sure that phase separation and enzyme activity are both not hindered.


Source

The amino acid sequence was taken from the publication by Tim Nott:

Nott TJ, Petsalaki E, Farber P, et al. Phase Transition of a Disordered Nuage Protein Generates Environmentally Responsive Membraneless Organelles. Molecular Cell. 2015;57(5):936-947. doi:10.1016/j.molcel.2015.01.013.

It was codon optimized for S.cerevisiae and we included bigger linkers.

References