Table 2.

List of plasmids used and generated in the study

Plasmid	Source	Description
pYG026	Addgene plasmid # 65328	Yeast ARS/CEN plasmid carrying the yeGFP and Leu2 and URA3 auxotrophic selection genes
pITGFP89	Addgene plasmid #83560	to amplify the yeGFP-CLN2 CDS
pLEP771	Dr. Kiran Madura	Yeast integrating plasmid used to tag Pup1 protein with tDimer red fluorescent protein
pFA6a-HIS3MX6	Dr. Marc Gartenberg	Used to amplify the spHIS3 gene with BAR1 overhangs to delete the BAR1 gene
pSD01	This study: synthesized the yeast-optimized d2eGFP (yeGFP-mODC) gene with GAL1 promoter on a bacterial cloning vector with ampicillin selection from Twist Biosciences.	To amplify the yeGFP-mODC CDS and GAL1 promoter sequence.
pSD02	This study	The variable promoter region swapped for GAL1 promoter amplified from pSD01. Used as the template downstream for creating the degron GFP constructs.
pSD03	This study	yeGFP-mODC CDS cloned into the pSD02 plasmid. Used to create constructs for genomic integration in SDY011.
pSD04	This study	yeGFP-CLN2 CDS cloned into the pSD02 plasmid. Used to create constructs for genomic integration in SDY011.
pSD09	This study	ARS/CEN and mCherry cassette deleted pSD03 plasmid linearized with BstEII for genomic integration of yeGFP-mODC at LEU2 locus.
pSD10	This study	ARS/CEN and mCherry cassette deleted pSD04 plasmid linearized with BstEII for genomic integration of yeGFP-CLN2 at LEU2 locus.