Plasmids used in this study
| Plasmid . | Backbone . | Insert . | Reference . |
|---|---|---|---|
| EGFP | pEGFP-N1 | None | |
| PH-AKT1-EGFP | pEGFP-N1 | AKT1(1–164):DPPVAT:EGFP | |
| PH-AKT1R25C-EGFP | pEGFP-N1 | AKT1(1–164)-R25C:DPPVAT:EGFP | |
| PH-PLCD1-mNeonGreen | pmNeonGreen-N1 | PLCD1(1–170):GVGG:mNeonGreen | |
| LynN11-FRB-iRFP | piRFP713-N1 | LYN(1–11):MTOR(2021–2113):iRFP713 | |
| LynN11-mNeonGreenx1 | pNeonGreen-N1 | LYN(1–11):GVGG:mNeonGreen | |
| LynN11-mNeonGreenx2 | LYN(1–11):GVGG:mNeonGreen:SPVAT:mNeonGreen | ||
| LynN11-mNeonGreenx3 | LYN(1–11):GVGG:mNeonGreen:SPVAT:mNeonGreen:SPVAT:mNeonGreen | ||
| mCherry-FKBP-iSH2 | mCherry: SGLRSRAALG:FKBP1A(3–108): SA[GGSA]4PRAQAS:Mus musculusPIK3R2(420–615) | Suh et al. (2006) | |
| PH-AKT1-mCherry | pmCherry-N1 | AKT1(1–164):DPPVAT:mCherry | |
| PH-BTK-mCherry | pmCherry-N1 | BTK(1–177):DPPVAT:mCherry | Várnai et al. (1999) |
| NES-iRFP670-aPHx1 | pCMVd3-iRFP670-C1a | X. leavis map2k1.L(32–44):PVAT:iRFP670: SGLRSRAQASNSAVDM:CYTH2i2(252–399)-I303E | Goulden et al. (2019) |
| NES-mCherry-aPHx2 | pCMVd3-mCherry-C1a | X. leavis map2k1.L(32–44):PVAT:mCherry: SGLRSRAQASNSAVDM:CYTH2i2(252–399)-I303E: GGGGGATCGGGTGGTGTCGACATG: CYTH2i2(252–399)-I303E | Goulden et al. (2019) |
| NES-iRFP670-cPHx1 | pCMVd3-iRFP670-C1a | X. leavis map2k1.L(32–44):PVAT:iRFP670: SGLRSRAGGAGAILS: PLEKHA1(169–329) | Goulden et al. (2019) |
| NES-mCherry-aPHx3 | pCMVd3-mCherry-C1a | X. leavis map2k1.L(32–44):PVAT:mCherry: SGLRSRAQASNSTWKMSS:PLEKHA1(169–329):GGSGGSGG: PLEKHA1(169–329): GGSGGSGG: PLEKHA1(169–329) | Goulden et al. (2019) |
| Plasmid . | Backbone . | Insert . | Reference . |
|---|---|---|---|
| EGFP | pEGFP-N1 | None | |
| PH-AKT1-EGFP | pEGFP-N1 | AKT1(1–164):DPPVAT:EGFP | |
| PH-AKT1R25C-EGFP | pEGFP-N1 | AKT1(1–164)-R25C:DPPVAT:EGFP | |
| PH-PLCD1-mNeonGreen | pmNeonGreen-N1 | PLCD1(1–170):GVGG:mNeonGreen | |
| LynN11-FRB-iRFP | piRFP713-N1 | LYN(1–11):MTOR(2021–2113):iRFP713 | |
| LynN11-mNeonGreenx1 | pNeonGreen-N1 | LYN(1–11):GVGG:mNeonGreen | |
| LynN11-mNeonGreenx2 | LYN(1–11):GVGG:mNeonGreen:SPVAT:mNeonGreen | ||
| LynN11-mNeonGreenx3 | LYN(1–11):GVGG:mNeonGreen:SPVAT:mNeonGreen:SPVAT:mNeonGreen | ||
| mCherry-FKBP-iSH2 | mCherry: SGLRSRAALG:FKBP1A(3–108): SA[GGSA]4PRAQAS:Mus musculusPIK3R2(420–615) | Suh et al. (2006) | |
| PH-AKT1-mCherry | pmCherry-N1 | AKT1(1–164):DPPVAT:mCherry | |
| PH-BTK-mCherry | pmCherry-N1 | BTK(1–177):DPPVAT:mCherry | Várnai et al. (1999) |
| NES-iRFP670-aPHx1 | pCMVd3-iRFP670-C1a | X. leavis map2k1.L(32–44):PVAT:iRFP670: SGLRSRAQASNSAVDM:CYTH2i2(252–399)-I303E | Goulden et al. (2019) |
| NES-mCherry-aPHx2 | pCMVd3-mCherry-C1a | X. leavis map2k1.L(32–44):PVAT:mCherry: SGLRSRAQASNSAVDM:CYTH2i2(252–399)-I303E: GGGGGATCGGGTGGTGTCGACATG: CYTH2i2(252–399)-I303E | Goulden et al. (2019) |
| NES-iRFP670-cPHx1 | pCMVd3-iRFP670-C1a | X. leavis map2k1.L(32–44):PVAT:iRFP670: SGLRSRAGGAGAILS: PLEKHA1(169–329) | Goulden et al. (2019) |
| NES-mCherry-aPHx3 | pCMVd3-mCherry-C1a | X. leavis map2k1.L(32–44):PVAT:mCherry: SGLRSRAQASNSTWKMSS:PLEKHA1(169–329):GGSGGSGG: PLEKHA1(169–329): GGSGGSGG: PLEKHA1(169–329) | Goulden et al. (2019) |
All genes (except fluorescent proteins) are human unless otherwise stated. Amino acid linkers are indicated with single letter code.
pCMVd3 plasmids are based on the pEGFP-C1 backbone, with the CMV promoter truncated to remove 18 of the 26 putative transcription factor–binding sites in the human cytomegalovirus major intermediate enhancer/promoter (pCMV∆3 as described in Morita et al. [2012]).