Table 2.
Plasmids used in this study
PlasmidDescriptionSource/reference
BG1805 2 µm, URA3, GAL1prom, C-terminal tandem affinity (TAP) tag vector Open Biosystems 
pMAL-c5X Bacterial expression vector for production of MBP (MalE) fusion proteins New England BioLabs 
pESC-Leu 2 µm, GAL1/10prom, LEU2 Agilent 
YCpLG CEN, LEU2, GAL1/10prom vector Bardwell et al., 1998  
pRS425 2 µm, LEU2 vector Sikorski and Hieter, 1989  
pRS426 2 µm, URA3 vector Sikorski and Hieter, 1989  
pRS315 CEN, LEU2 vector Sikorski and Hieter, 1989  
pET30a Bacterial expression vector Novagen 
pMLT35 pMAL-c5X MBP-MUK1-(His)6 This study 
pMLT74 pMAL-c5X MBP-muk1(S171A S172A S173A S183A S184A S185A)-(His)6 This study 
pMLT22 pESC-LEU GAL1prom-MUK1-Myc This study 
pMLT56 pESC-LEU GAL1prom-muk1(1-220)-Myc This study 
pMLT57 pESC-LEU GAL1prom-muk1(1-220; S171A S172A S173A S183A S184A S185A)-Myc This study 
pAX50 BG1805 YPK1(L424A)-TAP Muir et al., 2014  
pFR252 pRS315 ypk1(S51A T57A S71A T504A S644A S653A T662A)-Myc Muir et al., 2015  
pAM20 pRS315 Ypk1-myc Roelants et al., 2011  
pMLT101 pRS426 GAL1prom-FLAG-VPS21 This study 
pMLT102 pRS426 GAL1prom-FLAG-vps21(S21L) This study 
pMLT103 pRS426 GAL1prom-FLAG-vps21(Q66L) This study 
pMLT110 pRS426 GAL1prom-FLAG-YPT7 This study 
pMLT114 pRS426 GAL1prom-FLAG-mNG This study 
pMLT111 pRS426 MSB3-FLAG This study 
pMLT83 pRS315 MUK1-3xFLAG This study 
pMLT84 pRS315 muk1(S171A S172A S173A S183A S184A S185A)-3xFLAG This study 
pMLT85 pRS315 muk1(D353A)-3xFLAG This study 
pMLT92 pRS315 MUK1(S171E S172E S173E S183E S184E S185E)-3xFLAG This study 
pMLT132 pET30a ypk1(D242 D470A)-(His)6 This study 
pMLT135 pMAL-c5X MBP-Ypk1(603-680) [MBP-Ypk1CTThis study 
pMLT138 pMAL-c5X MBP-Ypk1(603-680; S644 S653A T662A S671A S672A S678) [MBP-Ypk1CT (6A)] This study 
pMLT88 pET30a VPS21-(His)6 This study 
pMLT141 pET30a YPT7-(His)6 This study 
pPL495 pRS315 PHSlm1-YPK1-3xHA Niles et al., 2012  
pMLT115 pRS425 GAL1prom-GST This study 
pML116 pRS425 GAL1prom-GST-Ypk1 This study 
PlasmidDescriptionSource/reference
BG1805 2 µm, URA3, GAL1prom, C-terminal tandem affinity (TAP) tag vector Open Biosystems 
pMAL-c5X Bacterial expression vector for production of MBP (MalE) fusion proteins New England BioLabs 
pESC-Leu 2 µm, GAL1/10prom, LEU2 Agilent 
YCpLG CEN, LEU2, GAL1/10prom vector Bardwell et al., 1998  
pRS425 2 µm, LEU2 vector Sikorski and Hieter, 1989  
pRS426 2 µm, URA3 vector Sikorski and Hieter, 1989  
pRS315 CEN, LEU2 vector Sikorski and Hieter, 1989  
pET30a Bacterial expression vector Novagen 
pMLT35 pMAL-c5X MBP-MUK1-(His)6 This study 
pMLT74 pMAL-c5X MBP-muk1(S171A S172A S173A S183A S184A S185A)-(His)6 This study 
pMLT22 pESC-LEU GAL1prom-MUK1-Myc This study 
pMLT56 pESC-LEU GAL1prom-muk1(1-220)-Myc This study 
pMLT57 pESC-LEU GAL1prom-muk1(1-220; S171A S172A S173A S183A S184A S185A)-Myc This study 
pAX50 BG1805 YPK1(L424A)-TAP Muir et al., 2014  
pFR252 pRS315 ypk1(S51A T57A S71A T504A S644A S653A T662A)-Myc Muir et al., 2015  
pAM20 pRS315 Ypk1-myc Roelants et al., 2011  
pMLT101 pRS426 GAL1prom-FLAG-VPS21 This study 
pMLT102 pRS426 GAL1prom-FLAG-vps21(S21L) This study 
pMLT103 pRS426 GAL1prom-FLAG-vps21(Q66L) This study 
pMLT110 pRS426 GAL1prom-FLAG-YPT7 This study 
pMLT114 pRS426 GAL1prom-FLAG-mNG This study 
pMLT111 pRS426 MSB3-FLAG This study 
pMLT83 pRS315 MUK1-3xFLAG This study 
pMLT84 pRS315 muk1(S171A S172A S173A S183A S184A S185A)-3xFLAG This study 
pMLT85 pRS315 muk1(D353A)-3xFLAG This study 
pMLT92 pRS315 MUK1(S171E S172E S173E S183E S184E S185E)-3xFLAG This study 
pMLT132 pET30a ypk1(D242 D470A)-(His)6 This study 
pMLT135 pMAL-c5X MBP-Ypk1(603-680) [MBP-Ypk1CTThis study 
pMLT138 pMAL-c5X MBP-Ypk1(603-680; S644 S653A T662A S671A S672A S678) [MBP-Ypk1CT (6A)] This study 
pMLT88 pET30a VPS21-(His)6 This study 
pMLT141 pET30a YPT7-(His)6 This study 
pPL495 pRS315 PHSlm1-YPK1-3xHA Niles et al., 2012  
pMLT115 pRS425 GAL1prom-GST This study 
pML116 pRS425 GAL1prom-GST-Ypk1 This study 

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