TABLE 1.

List of plasmids

PlasmidVectorInsertConstruction
YEp1338 HIS3 2μm pADH1-gal4(dbd)-ime1(270-360) Ligation
YIp1611 BS rim11Δ::LEU2 Ligation
YEp1788 LEU2 2μm pUME6-ume6(1-232)-gal4(ad)-ADHt Ligation
YIp1902 URA3 cdc25-5::URA3 Gap repair (32)
YEp1931 TRP1 2μm pIME1-gal4(dbd)-ime1(270-360) Ligation
YCp1955 HIS3 2μm CEN11 pAdh1-gal4(dbd) Ligation
YEp2042 TRP1 2μm hop1-lacZ Ligation
YEp2112 TRP1 2μm pIME1-gal4(dbd)-ime1S302A(270-360) Site-directed mutagenesis
YEp2122 TRP1 2μm pIME1-gal4(dbd)-ime1S306A(270-360) Site-directed mutagenesis
YIp2127 HIS3 rim11K68A Site-directed mutagenesis
YCp2195 HIS3 2μm CEN11 pADH1-gal4(dbd)-ime1Y359F(270-360) Random mutagenesis
YEp2196 HIS3 2μm pIME1-gal4(dbd)-ime1Y359F(270-360) Ligation
YEp2200 TRP1 2μm pIME1-gal4(dbd)-ime1(270-360) Ligation
YEp2206 TRP1 2μm pIME1-gal4(dbd)-ime1S333A(270-360) Site-directed mutagenesis
p2211T Easy rim15::loxp-URA3-kanr-loxp Ligation
YIp2214 LEU2 ime1Y359F(−1439 to +1081)-ADHt Ligation
YEp2215 TRP1 2μm pIME1-gal4(dbd)-ime1S337A(270-360) Site-directed mutagenesis
YCp2233 HIS3 ARSH4 + CEN6 pADH1-gal4(dbd)-ime1S302A(270-360) Ligation
YIp2234 LEU2 ime1(−1439 to +1081)-ADHt Ligation
YCp2242 HIS3 ARSH4 + CEN6 pADH1-gal4(dbd)-ime1S306A(270-360) Ligation
YCp2302 HIS3 ARSH4 + CEN6 pADH1-gal4(dbd)-ime1S302D(270-360) Ligation
YCp2303 HIS3 ARSH4 + CEN6 pADH1-gal4(dbd)-ime1S337A(270-360) Ligation
YCp2306 HIS3 ARSH4 + CEN6 pADH1-gal4(dbd)-ime1S333A(270-360) Ligation
YEp2430 TRP1 2μm pIME1-gal4(dbd)-ime1S352A,S356A,S360A (270-360) Site-directed mutagenesis
YCp2435 HIS3 ARSH4 + CEN6 pADH1-gal4(1-147)-ime1(270-360) Ligation
YCp2436 HIS3 ARSH4 + CEN6 pADH1-gal4(dbd)-ime1 S352A,S356A,S360A (270-360) Ligation
YIp2670 LEU2 Rim15K823A Ligation
P2677ptac-GST-Ime1(270-360)Ligation
YIp2691 LEU2 pRIM11-rim11K68A(1-223) Ligation
YEp2713 TRP1 2μm pIME1-gal4(dbd)-ime1S306,Y359F (270-360) Ligation
YEp2780 TRP1 2μm pADH1-gal4(dbd)-ime1(270-360) Ligation
YIp2804 URA3 Rim11-3SA Site-directed mutagenesis
P2843 ptac-gal4(92-147)-ime1(270-360)(Y359F) Ligation
P2844 pT7-RIM11-polyA Ligation