Strategy for N-degron GFP* reporter gene construction and integration into the S. cerevisiae genome. Plasmids with different UAS-UBI-X-ΔkGFP* reporters are constructed by combining UAS and GFP targeting cassettes. The X-Δk element of the reporter gene represents different bipartite N-degron signal sequences (see text). Digestion of the reporter gene plasmid with SacI and SalI restriction endonucleases releases the reporter gene from the plasmid backbone and generates free ends that target the allele to the URA3-TIM9 intergenic region of chromosome V. Unique restriction enzyme sites in the different cassettes are shown (B, BamHI; G, BglII; E, EcoRV; K, KpnI; Sa, SalI; Sc, SacI; Sm, SmaI; X, XhoI.)