Table 2. Plasmids used in this study... | Rockefeller University Press

Table 2.

Plasmids used in this study

Plasmid	Description^a	Reference or source
pGEX-2T	For expression of GST in E. coli	GE Healthcare
pGEX-RBD (PKC1)	For expression of GST-RBD (Pkc1) in E. coli	Yoshida et al., 2006; Kono et al., 2008
pGEX2T-CYK3	For expression of GST-Cyk3 in E. coli	See Supplemental material
pGEX2T-CYK3^480–885	For expression of GST-Cyk3^480-885 in E. coli	See Supplemental material
YEp13^b	2µ and LEU2	Rose and Broach, 1990
pJC328	CEN, URA3, and CHS2-MYC	Chuang and Schekman, 1996
YCp111-CDC3-CFP	CEN, LEU2, and CDC3-CFP	Nishihama et al., 2009
pRS316GW-IQG1	CEN, URA3, and IQG1	Nishihama et al., 2009
pRS316GW-HOF1	CEN, URA3, and HOF1	Unpublished data
pRS425GW-CYK3^b	2µ, LEU2, and CYK3	Unpublished data
pRS316GW-MYO1	CEN, URA3, and MYO1	Unpublished data
YEp13-RHO1^b	2µ, LEU2, and RHO1	This study^c
pGP564-RHO2^b	2µ, LEU2, RHO2, and neighboring genes^d	This study^e
YEp13-RGD2^b	2µ, LEU2, and RGD2	This study^c
YEp13-cdc24*^b^,^f	2µ, LEU2, and cdc24*	This study^c
YEp13-CDC24^b^,^f	2µ, LEU2, and CDC24	See Supplemental material
pSD1^b	2µ, LEU2, and MSS4	M.N. Hall^g
YEp13-KDX1^b	2µ, LEU2, and KDX1	This study^c
pGP564-PSP1^b	2µ, LEU2, PSP1, and neighboring genes^d	This study^e
YEp13-NAB6^b	2µ, LEU2, NAB6, and neighboring genes^d	This study^c
YEp13-PAP1^b	2µ, LEU2, and PAP1	This study^c
pRS425GW-TUS1^b	2µ, LEU2, and TUS1	See Supplemental material
pRS425GW-ROM1^b	2µ, LEU2, and ROM1	See Supplemental material
pRS425GW-ROM2^b	2µ, LEU2, and ROM2	See Supplemental material
pRS425GW-RGD1^b	2µ, LEU2, and RGD1	See Supplemental material
YEp181-RGA1^b	2µ, LEU2, and RGA1	Caviston et al., 2003
YEp181-RGA2^b	2µ, LEU2, and RGA2	See Supplemental material
YEp181-BEM2^b	2µ, LEU2, and BEM2	Unpublished data
YEp13-BEM3^b	2µ, LEU2, and BEM3	Bi and Pringle, 1996
pRS425GW-SAC7^b	2µ, LEU2, and SAC7	See Supplemental material
pRS425GW-LRG1^b	2µ, LEU2, and LRG1	See Supplemental material
pGP564-BAG7^b	2µ, LEU2, BAG7, and neighboring genes^d	This study^e
YEp13-cdc24ΔPB1^f	2µ, LEU2, and cdc24ΔPB1	See Supplemental material
YEp13-cdc24*-DH8^f	2µ, LEU2, and cdc24* with N452G and E453G mutations	See Supplemental material
pRS425-CDC24-GFP^f	2µ, LEU2, and CDC24-GFP	See Supplemental material
YEp13-cdc24*-GFP^f	2µ, LEU2, and cdc24ΔPB1-GFP-tetA	See Supplemental material
pRS425-cdc24ΔPB1-GFP^f	2µ, LEU2, and cdc24ΔPB1-GFP	See Supplemental material
YCp-P_GAL-RHO1^T24N	CEN, LEU2, and P_GAL-RHO1^T24N	See Supplemental material
YIp128-P_GAL-RHO1^Q68L	Integrative, LEU2, and P_GAL-RHO1^Q68L	See Supplemental material
pRS306-3HA-RHO1	Integrative, URA3, and 3HA-RHO1	See Supplemental material
YCp-P_GAL-3HA-RHO1	CEN, LEU2, and P_GAL-3HA-RHO1	See Supplemental material
YCp-P_GAL-3HA-RHO1^Q68L	CEN, LEU2, and P_GAL-3HA-RHO1^Q68L	See Supplemental material
YCp-P_GAL-3HA-RHO1^T24N	CEN, LEU2, and P_GAL-3HA-RHO1^T24N	See Supplemental material
pFA6a-VN173:TRP1	PCR template for chromosomal tagging with VN173	Sung and Huh, 2007
pFA6a-VN173:His3MX6	PCR template for chromosomal tagging with VN173	Sung and Huh, 2007
pFA6a-VC155-TRP1	PCR template for chromosomal tagging with VC155	Sung and Huh, 2007
pFA6a-VN155^I153L-TRP1	PCR template for chromosomal tagging with VN155^I153L	See Supplemental material
pRS315-VC-GGS₂-RHO1	CEN, LEU2, and VC155-GGSGGS-RHO1	See Supplemental material
pRS315-CYK3^TGcΔ^h	CEN, LEU2, and CYK3 lacking bp 1,556–1,739	See Supplemental material

Plasmid	Description^a	Reference or source
pGEX-2T	For expression of GST in E. coli	GE Healthcare
pGEX-RBD (PKC1)	For expression of GST-RBD (Pkc1) in E. coli	Yoshida et al., 2006; Kono et al., 2008
pGEX2T-CYK3	For expression of GST-Cyk3 in E. coli	See Supplemental material
pGEX2T-CYK3^480–885	For expression of GST-Cyk3^480-885 in E. coli	See Supplemental material
YEp13^b	2µ and LEU2	Rose and Broach, 1990
pJC328	CEN, URA3, and CHS2-MYC	Chuang and Schekman, 1996
YCp111-CDC3-CFP	CEN, LEU2, and CDC3-CFP	Nishihama et al., 2009
pRS316GW-IQG1	CEN, URA3, and IQG1	Nishihama et al., 2009
pRS316GW-HOF1	CEN, URA3, and HOF1	Unpublished data
pRS425GW-CYK3^b	2µ, LEU2, and CYK3	Unpublished data
pRS316GW-MYO1	CEN, URA3, and MYO1	Unpublished data
YEp13-RHO1^b	2µ, LEU2, and RHO1	This study^c
pGP564-RHO2^b	2µ, LEU2, RHO2, and neighboring genes^d	This study^e
YEp13-RGD2^b	2µ, LEU2, and RGD2	This study^c
YEp13-cdc24*^b^,^f	2µ, LEU2, and cdc24*	This study^c
YEp13-CDC24^b^,^f	2µ, LEU2, and CDC24	See Supplemental material
pSD1^b	2µ, LEU2, and MSS4	M.N. Hall^g
YEp13-KDX1^b	2µ, LEU2, and KDX1	This study^c
pGP564-PSP1^b	2µ, LEU2, PSP1, and neighboring genes^d	This study^e
YEp13-NAB6^b	2µ, LEU2, NAB6, and neighboring genes^d	This study^c
YEp13-PAP1^b	2µ, LEU2, and PAP1	This study^c
pRS425GW-TUS1^b	2µ, LEU2, and TUS1	See Supplemental material
pRS425GW-ROM1^b	2µ, LEU2, and ROM1	See Supplemental material
pRS425GW-ROM2^b	2µ, LEU2, and ROM2	See Supplemental material
pRS425GW-RGD1^b	2µ, LEU2, and RGD1	See Supplemental material
YEp181-RGA1^b	2µ, LEU2, and RGA1	Caviston et al., 2003
YEp181-RGA2^b	2µ, LEU2, and RGA2	See Supplemental material
YEp181-BEM2^b	2µ, LEU2, and BEM2	Unpublished data
YEp13-BEM3^b	2µ, LEU2, and BEM3	Bi and Pringle, 1996
pRS425GW-SAC7^b	2µ, LEU2, and SAC7	See Supplemental material
pRS425GW-LRG1^b	2µ, LEU2, and LRG1	See Supplemental material
pGP564-BAG7^b	2µ, LEU2, BAG7, and neighboring genes^d	This study^e
YEp13-cdc24ΔPB1^f	2µ, LEU2, and cdc24ΔPB1	See Supplemental material
YEp13-cdc24*-DH8^f	2µ, LEU2, and cdc24* with N452G and E453G mutations	See Supplemental material
pRS425-CDC24-GFP^f	2µ, LEU2, and CDC24-GFP	See Supplemental material
YEp13-cdc24*-GFP^f	2µ, LEU2, and cdc24ΔPB1-GFP-tetA	See Supplemental material
pRS425-cdc24ΔPB1-GFP^f	2µ, LEU2, and cdc24ΔPB1-GFP	See Supplemental material
YCp-P_GAL-RHO1^T24N	CEN, LEU2, and P_GAL-RHO1^T24N	See Supplemental material
YIp128-P_GAL-RHO1^Q68L	Integrative, LEU2, and P_GAL-RHO1^Q68L	See Supplemental material
pRS306-3HA-RHO1	Integrative, URA3, and 3HA-RHO1	See Supplemental material
YCp-P_GAL-3HA-RHO1	CEN, LEU2, and P_GAL-3HA-RHO1	See Supplemental material
YCp-P_GAL-3HA-RHO1^Q68L	CEN, LEU2, and P_GAL-3HA-RHO1^Q68L	See Supplemental material
YCp-P_GAL-3HA-RHO1^T24N	CEN, LEU2, and P_GAL-3HA-RHO1^T24N	See Supplemental material
pFA6a-VN173:TRP1	PCR template for chromosomal tagging with VN173	Sung and Huh, 2007
pFA6a-VN173:His3MX6	PCR template for chromosomal tagging with VN173	Sung and Huh, 2007
pFA6a-VC155-TRP1	PCR template for chromosomal tagging with VC155	Sung and Huh, 2007
pFA6a-VN155^I153L-TRP1	PCR template for chromosomal tagging with VN155^I153L	See Supplemental material
pRS315-VC-GGS₂-RHO1	CEN, LEU2, and VC155-GGSGGS-RHO1	See Supplemental material
pRS315-CYK3^TGcΔ^h	CEN, LEU2, and CYK3 lacking bp 1,556–1,739	See Supplemental material

CEN indicates a low-copy number plasmid; 2µ indicates a high-copy number plasmid; integrative indicates a plasmid without a yeast replication origin.

Used in Fig. 2 A and/or Fig. S1.

Isolated in the suppressor screen using the YEp13-based library (see Materials and methods) or by subcloning a single-gene fragment from the originally isolated plasmid back into YEp13.

The actual plasmids used in the experiments shown in this paper are listed. The genes responsible for suppression were confirmed by further truncation.

Isolated from a genomic tiling library (Thermo Fisher Scientific).

Used in Fig. S3.

University of Basel, Basel, Switzerland.

Used for construction of strain MOY882.

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