We used pEGFP-N1 (https://www.addgene.org/vector-database/2491/) as a backbone vector. Lyn, cytoplasmic domain of FAS and PHR domain were sequentially inserted to the backbone vector by restriction enzyme cloning. 

First, Lyn sequence was PCR-amplified from optoFGFR1 (https://www.addgene.org/59776/) by using the following primers and inserted to pEGFP-N1 with NheI/XhoI digestion to construct Lyn-EGFP: 5′-GTAGCTAGCCACC ATGGGATGTATAAAATCAAAAGG-3′ (forward) and 5′-GTACTCGAGCGCACTACCAGCACTACCAG-3′ (reverse). 

Then, cytoplasmic domain of FAS was PCR-amplified from HeLa cell cDNA by using following primers and inserted to Lyn-EGFP with XhoI/EcoRI digestion to construct Lyn-cyFAS-EGFP: 5′-GTACTCGAGAAGAGAAAGGAAGTACAGAAAACATGCAGA-3′ (forward) and 5′-GTAGAATTCTGACCAAGCTTTGGATTTCATTT-3′ (reverse). 

Finally, PHR domain was PCR-amplified from optoFGFR1 (https://www.addgene.org/59776/) by using the following primers and inserted to Lyn-cyFAS-EGFP with BamHI/AgeI digestion to construct Lyn-cyFAS-PHR-EGFP (optoFAS): 5′-GTAGGATCCCATGAAGATGGACAAAAAGACCA-3′ (forward) and 5′-GTAACCGGTGCGTACACGGCAGCACCGATC-3′ (reverse).