REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Anti-Flag M2 magnetic beads	Sigma-Aldrich	M8823
Anti-Flag M2 mouse	Sigma-Aldrich	P2983, RRID:AB_439685
Anti-Flag M2 affinity gel	Sigma-Aldrich	A2220
Anti-Sui2 (rabbit)	Dr T. Dever, NIH
Anti-Gcn3	Dr A. Hinnebusch, NIH
Anti-Gcd7	Dr A. Hinnebusch, NIH
Anti-Gcd6	Dr A. Hinnebusch, NIH
Anti-Gcd2	Dr A. Hinnebusch, NIH
Anti-Tef1	Dr C. Grant, Univ. Manchester UK
IRDye® 800CW goat anti-rabbit IgG	LI-COR Biosciences	926-32211, RRID:AB_621843
IRDye® 680RD goat anti-mouse IgG	LI-COR Biosciences	925-68070, RRID:AB_2651128
Bacterial and virus strains
E. coli XL1-Blue Supercompetent Cells	Agilent	200523
Chemicals, peptides, and recombinant proteins
Complete Protease Inhibitor mini tablets, EDTA free	Roche	11836170001
Yeast Nitrogen Base Without Amino Acids	Formedium	CYN0402
SC, Double Drop-Out -Leu, -Ura	Formedium	DSCK1017
5-Fluoro Orotic Acid Monohydrate	Formedium	5FOA05
3-Amino-1,2,4-triazole	Sigma-Aldrich	A8056
Adenine	Sigma-Aldrich	A8626
Guanine	Sigma-Aldrich	G11950
Guanosine 5′-triphosphate sodium salt hydrate	Sigma-Aldrich	G8877
Guanosine 5′-diphosphate sodium salt	Sigma-Aldrich	G7127
Adenosine 5′-diphosphate sodium salt	Sigma-Aldrich	A2754
Guanosine 5'-Diphosphate, BODIPY™ FL 2'-(or-3')-O-(N-(2-Aminoethyl) Urethane), Bis (Triethylammonium) Salt)	ThermoFisher	G22360
GTP, [α-32P]- 3000Ci/mmol 10mCi/ml	PerkinElmer	BLU006H250UC
Ultima Gold F	PerkinElmer	6013179
Critical commercial assays
QuikChange II site direct mutagenesis	Agilent	200523
NiNTA agarose	Qiagen	30210
HiTrap heparin	Cytiva	17-0407-01
HiTrap Q	Cytiva	17-1153-01
Experimental models: Organisms/strains
Saccharomyces cerevisiae strains derived from S288c	Lab collection	see Table S1
Oligonucleotides
GCGCTTGCCCGCGGCTCTTTTGCCCATCGGTAATAG	Sigma-Aldrich	K66A F
GCAAAAGAGCCGCGGGCAAGCGCGTTGAATGTTGC	Sigma-Aldrich	K66A R
GCGCTTGCCCGACGCTCTTTTGCCCATCGGTAATAG	Sigma-Aldrich	K66D F
GCAAAAGAGCGTCGGGCAAGCGCGTTGAATGTTGC	Sigma-Aldrich	K66D R
GCGCTTGCCCAGGGCTCTTTTGCCCATCGGTAATAG	Sigma-Aldrich	K66R F
GCAAAAGAGCCCTGGGCAAGCGCGTTGAATGTTGC	Sigma-Aldrich	K66R R
CTTGCCCTGCGCATTTGTCACAGATATACCTCCACAAGTC	Sigma-Aldrich	D173A F
CTGTGACAAATGCGCAGGGCAAGATTACAAAATCGCCATTG	Sigma-Aldrich	D173A R
CTTGCCCTGTAAGTTTGTCACAGATATACCTCCACAAGTC	Sigma-Aldrich	D173K F
CTGTGACAAACTTACAGGGCAAGATTACAAAATCGCCATTG	Sigma-Aldrich	D173K R
CTTGCCCTGTAATTTTGTCACAGATATACCTCCACAAGTC	Sigma-Aldrich	D173N F
CTGTGACAAAATTACAGGGCAAGATTACAAAATCGCCATTG	Sigma-Aldrich	D173N R
GACAGACTCTTATGAAACTAAATTTATGCCAC	Sigma-Aldrich	R39K F
CAGCTGTCAGTGGCATAAATTTAGTTTCATAAG	Sigma-Aldrich	R39K R
GACAGACTCTTATGAAAACTGATTTTATGCCAC	Sigma-Aldrich	R39D F
CAGCTGTCAGTGGCATAAAATCAGTTTCATAAG	Sigma-Aldrich	R39D R
GACAGACTCTTATGAAACTGAATTTATGCCAC	Sigma-Aldrich	R39E F
CAGCTGTCAGTGGCATAAATTCAGTTTCATAAG	Sigma-Aldrich	R39E R
GATTTTATTTTAGTCAGTGGTAATGTATTGACTAAC	Sigma-Aldrich	D138N F
GCTGAAATCGATGTTAGTCAATACATTACCACTGAC	Sigma-Aldrich	D138N R
GATTTTATTTTAGTCAGTGGTAAAGTATTGACTAAC	Sigma-Aldrich	D138K F
GCTGAAATCGATGTTAGTCAATACTTTACCACTGAC	Sigma-Aldrich	D138K R
Recombinant DNA
Yeast expression plasmids	This study or cited references	See Table S2
Software and algorithms
UCSF Chimera 1.15		(Pettersen et al., 2004)
homology modelling server SWISS-MODEL		(Waterhouse et al., 2018)
PISA		(Krissinel and Henrick, 2007)
FluorEssence	Horiba Scientific
ImageStudio	Li-Cor BioSciences