Construct was generated using Gibson assembly (Gibson et al., 2009), and verified by sequencing of new junction regions. Psun-1::TIR1::mRuby::sun-1
3'UTR was derived from pCFJ178. A codon-optimized TIR1 gene with two introns was assembled from multiple gBlocks (IDT). This coding sequence was used to replace the sun-1
coding sequence in pOR131 (Rog and Dernburg, 2015), a derivative of pCFJ178 containing the sun-1
promoter and coding sequence inserted upstream of a codon-optimized mRuby gene, followed by the sun-1
3'UTR. In the resulting construct, TIR1 and mRuby are separated by a 3x(Gly-Gly-Ser-Gly) linker.