We fuse part BBa_K209496 (Frb) and BBa_K209023 (FKBP) with yEGFP, which make it visible in the yeast under fluorescent microscope. Then we got partBBa_K2601008(FKBP-yEGFP) and BBa_K2601007(Frb-yEGFP).
Afterward, we drive the expression of FKBP-yEGFP and Frb-yEGFP with 4 promoters pUra3, pTet07,pTEF1 and PDH3. The expression of these promoters were measure by flow cytometry(Fig. 1). We then got prats BBa_K2601021 (Tet07-Frb-yEGFP), K2601023 (PDH3-Frb-yEGFP), BBa_K2601025 (Tet07-FKBP-yEGFP), BBa_K2601026 (TEF1-FKBP-yEGFP), K2601027 (PDH3-FKBP-yEGFP).
What's more, we fused Frb-yEGFP with HOtag6 and fused FKBP-yEGFP with HOtag3. These two parts can form phase separation in the presence of rapamycin (Fig. 2). The original part BBa_K209496 (Frb) and BBa_K209023 (FKBP) have not this function. We uploaded 2 parts: BBa_K2601010 (Frb-yEGFP-HOTag6) and BBa_K2601011 (FKBP-yEGFP-HOTag3).