Table 2.
Plasmids used in this study. 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.
| Shorthand Name | Sequence | Reference |
|---|---|---|
| NES-PABD-Spo20 | X.leavis 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.leavis map2k1.L32–44-APVAT-EGFP-SGGGSGGS-S. cerevisiae Spo20p51–91 | This Study |
| NES-PABDx2-Spo20 | X.leavis map2k1.L32–44-APVAT-EGFP-SGLRSRA-S. cerevisiae Spo20p51–91-S. cerevisiae Spo20p51–91 | (Bohdanowicz et al., 2013) |
| NESx2-PABDx2-Spo20 | X.leavis map2k1.L32–44-AGGSG-X.leavis 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.leavis 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.leavis 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., 2023) |
| 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., 2014) |
| 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) |