GFP-Cdc42 was expressed from the PCDC42-GFP-linker-CDC42 (linker: APPRRLVHP) targeted to the URA3 locus (using YIplac211-PCDC42-GFP-linker-CDC42). This strain and another strain that carries the extra copies of GFP-CDC42 were originally described in Kuo et al. (2014) Cure Biol 24:753–759. The endogenous CDC42 was deleted by crossing with DLY13891 (MATa cdc42::TRP1 URA3:GFP-CDC42 (8x) in the same genetic background) (Wu et al., 2015, eLife 4, 11611). As cited in the text (see the details in Wu et al., 2015), these fusions are not fully functional at a temperature higher than 34oC.
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