Our submitted plasmid contains both GAL4-KaiC fusion protein and a KaiB protein, linked by a self-cleaving linker:
The plasmid we submitted to the iGEM registry contains KaiC fused to a Gal4 activation domain, with a self-cleaving peptide linker, P2A, fusing it to KaiB. P2A cleaves itself during translation, causing the KaiC-Gal4AD and the KaiB to separate. We did this to allow us to control the exact ratio of KaiC, KaiB, and KaiA, and make sure it is constant and equal in every cell. It is also codon optimized for yeast, and contains an NLS sequence to localize it to the nucleus. This part was synthesized by Genscript.