Table 2.

Plasmids used in this study

Shorthand nameSequenceReference
NES-PABD-Spo20 X. laevis map2k1.L32-44-APVAT-EGFP-GLRSRASI-S. cerevisiae Spo20p51-91 Zeniou-Meyer et al. (2007), Zhang et al. (2014)  
PASS EGFP-SGLRSRA-M. musculus PKIa34-51-SR-S. cerevisiae Spo20p51-91 Zhang et al. (2014), Zeniou-Meyer et al. (2007)  
NES-flex-PABD-Spo20 X. laevis map2k1.L32-44-APVAT-EGFP-SGGGSGGS-S. cerevisiae Spo20p51-91 This study 
NES-PABDx2-Spo20 X. laevis map2k1.L32-44-APVAT-EGFP-SGLRSRA-S. cerevisiae Spo20p51-91- S. cerevisiae Spo20p51-91 Bohdanowicz et al. (2013)  
NESx2-PABDx2-Spo20 X. laevis map2k1.L32-44-AGGSG-X. laevis map2k1.L32-44-APVAT-EGFP-SGLRSRA-S. cerevisiae Spo20p51-91- S. cerevisiae Spo20p51-91 This study 
PILS-Nir1 (Nir1-LNS2) NeonGreen-GGSGGM-(PITPNM3) Nir1613-897 This study 
Nir2-LNS2 NeonGreen-GGSGGM-(PITPNM1) Nir2896-1216 This study 
Nir3-LNS2 pcDNA3.1-mEGFP-GGGGSHM-(PITPNM2 isoform 6) Nir3925-1209 This study 
PILS-Nir1-613-630 (PITPNM3) Nir1613-630-GGSGG-NeonGreen This study 
PILS-Nir1-631-894 NeonGreen-GGSGG-(PITPNM3) Nir1631-894 This study 
PILS-Nir1-LNS2-K820E NeonGreen-GGSGGM-(PITPNM3) Nir1613-897, K820E This study 
C1ab-Prkd1 X. laevis map2k1.L32-44-APVAT-mCherry-SGLRSRAQASNSTS-M. musculus Prkd1138-343 Kim et al. (2011)  
FKBP-PJ-dead mCherry-SGLRSRSAAAGAGGAARAALG-FKBP1A3-109-SAGGSAGGSAGGSAGGSAGGPRAQASRSLDA-S. cerevisiae Sac12-517, C392S -GGTARGAAAGAGGAGR-INPP5E214-644, D556A,C641A Hammond et al. (2012)  
FKBP-PJ mCherry-SGLRSRSAAAGAGGAARAALG-FKBP1A3-109-SAGGSAGGSAGGSAGGSAGGPRAQASRSLDA-S. cerevisiae Sac12-517-GGTARGAAAGAGGAGR-INPP5E214-644, C641A Hammond et al. (2012)  
FKBP-INPP5E mCherry-SGLRSRSAAAGAGGAARAAMG-FKBP1A3-109-ARGAAAGAGGAGR-INPP5E214-644, C641A Hammond et al. (2012)  
FKBP-PJ-Sac1 mCherry-SGLRSRSAAAGAGGAARAALG-FKBP1A3-109-SAGGSAGGSAGGSAGGSAGGPRAQASRSLDA-S. cerevisiae Sac12-517-GGTARGAAAGAGGAGR-INPP5E214-644, D556A C641A Hammond et al. (2012)  
BFP-Tubby(C) M. musculus Tubby protein243-505-TVPRARDA-pTagBFP-KRPRL Quinn et al. (2008)  
P4Mx1 X. laevis map2k1.L32-44-APVAT-mTagBFP2-SGLRSRAQASNSAVDGGSASGLRS- L. penomophila SidM546-647 Zewe et al. (2020)  
PM-FRB Lyn1-11-RSANSGAGAGAGAILSR-MTOR2021-2113-TSYPYDVPDYAPVAT-iRFP Hammond et al. (2014)  
Mito-FRB iRFP-SGLRSRAGGAGAILSR-MTOR2021-2113-GGSAGGSAQASNSAVDGTA-Fis1122-152 Doyle et al. (2024), Preprint 
FKBP-PI4K mCherry-SGLRSRSAAAGAGGAARAAL-FKBP1A3-108-SAGGSAGGSAGGSAGGSAGGPRAQASNSL-PI4KA1102-2103 Zewe et al. (2020)  
FKBP-PIP5K pTagBFP-SGLRSRSAAAGAGGAARAALG-FKBP1A3-108-SAGGSAGGSAGGSAGGSAGGPRAQASNSAVDLQA-PIP5K1C1-640,D101R, R304D, R445E, K446E This study 
EGFP-PH-PLCδ1 PLCδ11-170-DPPVAT-EGFP Várnai and Balla (1998)  
FKBP-PI-PLC B. cereus PI-PLC32-329, W78A, W273A-RILQSTVPMG-FKBP1A3-108-RDPPVATM-TagBFP2-SGLRSRSAAATLDHNQPYHICRGFTCFKKPPTPPPEPET Pemberton et al. (2020)  
FKBP-DGKa mRFP-SGLRSRSAAAGAGGAARAAL-FKBP1A3-108-SAGGSAGGSAGGSAGGSAGGPRAQASRS-DGKA isoform b394-773 This study 
M3 pcDNA3.1-HAx3-AchR-CHRM32-590 J. Wess 
ER-FRB iRFP713- SGLRSRAQLTMAYPYDVPDYVA-MTOR2021-2113- QGSGAGAGAGAILNSRV-SACM1L418-484 Zewe et al. (2020)  
Golgi-FRB iRFP-SGLRSRAGGAGAILSR-MTOR2021-2113-GGSAGGSA-GOLGB13096-3224 Zewe et al. (2020)  
Rab5-FRB iRFP713-SGLRSRAGGAGAILSR-MTOR2021-2113-GGSAGGSAQASNSAVDGT-C. lupus Rab5a1-215 Hammond et al. (2012)  
GFP-Nir2 EGFP-SGLRSRAQASNS-PITPNM1v2 Kim et al. (2015)  
iRFP-PILS-Nir1 miRFP670-GGSGGM-(PITPNM3) Nir1613-897 This study 
iRFP-Tubby(c) M. musculus Tubby243-505-PRARDPPVAT-miRFP670 Quinn et al. (2008)  
iRFP-PH-PLCδ1 iRFP713-CTRDLELKL-R. norvegicus PLCD1 isoform X32-131 Idevall-Hagren et al. (2012)  
TagBFP2-HRAS-CAAX TagBFP2-SGLRSRAQASNSAVD-HRAS172-189 Goulden et al. (2019)  
mCh-HRAS-CAAX mCherry-SGLRSRAQASNSAVD-HRAS172-189 This study 
pUC19 Empty plasmid used to bring total DNA mass up to 1 μg as needed Yanisch-Perron et al. (1985)  
Shorthand nameSequenceReference
NES-PABD-Spo20 X. laevis map2k1.L32-44-APVAT-EGFP-GLRSRASI-S. cerevisiae Spo20p51-91 Zeniou-Meyer et al. (2007), Zhang et al. (2014)  
PASS EGFP-SGLRSRA-M. musculus PKIa34-51-SR-S. cerevisiae Spo20p51-91 Zhang et al. (2014), Zeniou-Meyer et al. (2007)  
NES-flex-PABD-Spo20 X. laevis map2k1.L32-44-APVAT-EGFP-SGGGSGGS-S. cerevisiae Spo20p51-91 This study 
NES-PABDx2-Spo20 X. laevis map2k1.L32-44-APVAT-EGFP-SGLRSRA-S. cerevisiae Spo20p51-91- S. cerevisiae Spo20p51-91 Bohdanowicz et al. (2013)  
NESx2-PABDx2-Spo20 X. laevis map2k1.L32-44-AGGSG-X. laevis map2k1.L32-44-APVAT-EGFP-SGLRSRA-S. cerevisiae Spo20p51-91- S. cerevisiae Spo20p51-91 This study 
PILS-Nir1 (Nir1-LNS2) NeonGreen-GGSGGM-(PITPNM3) Nir1613-897 This study 
Nir2-LNS2 NeonGreen-GGSGGM-(PITPNM1) Nir2896-1216 This study 
Nir3-LNS2 pcDNA3.1-mEGFP-GGGGSHM-(PITPNM2 isoform 6) Nir3925-1209 This study 
PILS-Nir1-613-630 (PITPNM3) Nir1613-630-GGSGG-NeonGreen This study 
PILS-Nir1-631-894 NeonGreen-GGSGG-(PITPNM3) Nir1631-894 This study 
PILS-Nir1-LNS2-K820E NeonGreen-GGSGGM-(PITPNM3) Nir1613-897, K820E This study 
C1ab-Prkd1 X. laevis map2k1.L32-44-APVAT-mCherry-SGLRSRAQASNSTS-M. musculus Prkd1138-343 Kim et al. (2011)  
FKBP-PJ-dead mCherry-SGLRSRSAAAGAGGAARAALG-FKBP1A3-109-SAGGSAGGSAGGSAGGSAGGPRAQASRSLDA-S. cerevisiae Sac12-517, C392S -GGTARGAAAGAGGAGR-INPP5E214-644, D556A,C641A Hammond et al. (2012)  
FKBP-PJ mCherry-SGLRSRSAAAGAGGAARAALG-FKBP1A3-109-SAGGSAGGSAGGSAGGSAGGPRAQASRSLDA-S. cerevisiae Sac12-517-GGTARGAAAGAGGAGR-INPP5E214-644, C641A Hammond et al. (2012)  
FKBP-INPP5E mCherry-SGLRSRSAAAGAGGAARAAMG-FKBP1A3-109-ARGAAAGAGGAGR-INPP5E214-644, C641A Hammond et al. (2012)  
FKBP-PJ-Sac1 mCherry-SGLRSRSAAAGAGGAARAALG-FKBP1A3-109-SAGGSAGGSAGGSAGGSAGGPRAQASRSLDA-S. cerevisiae Sac12-517-GGTARGAAAGAGGAGR-INPP5E214-644, D556A C641A Hammond et al. (2012)  
BFP-Tubby(C) M. musculus Tubby protein243-505-TVPRARDA-pTagBFP-KRPRL Quinn et al. (2008)  
P4Mx1 X. laevis map2k1.L32-44-APVAT-mTagBFP2-SGLRSRAQASNSAVDGGSASGLRS- L. penomophila SidM546-647 Zewe et al. (2020)  
PM-FRB Lyn1-11-RSANSGAGAGAGAILSR-MTOR2021-2113-TSYPYDVPDYAPVAT-iRFP Hammond et al. (2014)  
Mito-FRB iRFP-SGLRSRAGGAGAILSR-MTOR2021-2113-GGSAGGSAQASNSAVDGTA-Fis1122-152 Doyle et al. (2024), Preprint 
FKBP-PI4K mCherry-SGLRSRSAAAGAGGAARAAL-FKBP1A3-108-SAGGSAGGSAGGSAGGSAGGPRAQASNSL-PI4KA1102-2103 Zewe et al. (2020)  
FKBP-PIP5K pTagBFP-SGLRSRSAAAGAGGAARAALG-FKBP1A3-108-SAGGSAGGSAGGSAGGSAGGPRAQASNSAVDLQA-PIP5K1C1-640,D101R, R304D, R445E, K446E This study 
EGFP-PH-PLCδ1 PLCδ11-170-DPPVAT-EGFP Várnai and Balla (1998)  
FKBP-PI-PLC B. cereus PI-PLC32-329, W78A, W273A-RILQSTVPMG-FKBP1A3-108-RDPPVATM-TagBFP2-SGLRSRSAAATLDHNQPYHICRGFTCFKKPPTPPPEPET Pemberton et al. (2020)  
FKBP-DGKa mRFP-SGLRSRSAAAGAGGAARAAL-FKBP1A3-108-SAGGSAGGSAGGSAGGSAGGPRAQASRS-DGKA isoform b394-773 This study 
M3 pcDNA3.1-HAx3-AchR-CHRM32-590 J. Wess 
ER-FRB iRFP713- SGLRSRAQLTMAYPYDVPDYVA-MTOR2021-2113- QGSGAGAGAGAILNSRV-SACM1L418-484 Zewe et al. (2020)  
Golgi-FRB iRFP-SGLRSRAGGAGAILSR-MTOR2021-2113-GGSAGGSA-GOLGB13096-3224 Zewe et al. (2020)  
Rab5-FRB iRFP713-SGLRSRAGGAGAILSR-MTOR2021-2113-GGSAGGSAQASNSAVDGT-C. lupus Rab5a1-215 Hammond et al. (2012)  
GFP-Nir2 EGFP-SGLRSRAQASNS-PITPNM1v2 Kim et al. (2015)  
iRFP-PILS-Nir1 miRFP670-GGSGGM-(PITPNM3) Nir1613-897 This study 
iRFP-Tubby(c) M. musculus Tubby243-505-PRARDPPVAT-miRFP670 Quinn et al. (2008)  
iRFP-PH-PLCδ1 iRFP713-CTRDLELKL-R. norvegicus PLCD1 isoform X32-131 Idevall-Hagren et al. (2012)  
TagBFP2-HRAS-CAAX TagBFP2-SGLRSRAQASNSAVD-HRAS172-189 Goulden et al. (2019)  
mCh-HRAS-CAAX mCherry-SGLRSRAQASNSAVD-HRAS172-189 This study 
pUC19 Empty plasmid used to bring total DNA mass up to 1 μg as needed Yanisch-Perron et al. (1985)  

All genes are Homo sapiens unless indicated otherwise. The relevant amino acid positions in the full-length protein are noted. Mutations are described by the position of the residues in the full-length protein. Amino acid sequences written out indicate linkers.

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