Part:BBa_K2483008:Design
IAA producing genes fused to droplet forming Ddx4 for S.cerevisiae
- 10INCOMPATIBLE WITH RFC[10]Illegal SpeI site found at 13
- 12INCOMPATIBLE WITH RFC[12]Illegal SpeI site found at 13
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1089
Illegal BamHI site found at 1302
Illegal BamHI site found at 4585
Illegal BamHI site found at 5989 - 23INCOMPATIBLE WITH RFC[23]Illegal SpeI site found at 13
- 25INCOMPATIBLE WITH RFC[25]Illegal SpeI site found at 13
Illegal NgoMIV site found at 5032
Illegal AgeI site found at 89
Illegal AgeI site found at 3409
Illegal AgeI site found at 5328 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 712
Illegal BsaI site found at 2110
Illegal BsaI site found at 5331
Illegal BsaI.rc site found at 665
Design Notes
Because our other parts (BBa_K2483007 and BBa_K2483009) for this project were also under control of the GAL1 promoter (strong, galactose-induced promoter), we needed to do the same here for it to be a control.
We did not check for illegal restricition sites, so the SpeI sites should have to be mutated out.
Source
The CDS for IAAM and IAAH are from the part BBa_K515100 and codon optimized for yeast. GAL1 and CYC1 are taken from the vector we were working with, which was pYES2/CT.
The Ddx4 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.