CDT1 inhibits CMG helicase in early S phase to separate origin licensing from DNA synthesis.
Ratnayeke N, Baris Y, Chung M, Yeeles JTP, Meyer T
Mol Cell. 2023 Jan 5;83(1):26-42.e13. doi: 10.1016/j.molcel.2022.12.004.
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Plasmids from Article
| ID | Plasmid | Purpose |
|---|---|---|
| 193139 | pLV-hCdt1(1-100)ΔCy-mCherry-P2A-mVenus-hGeminin(1-110)-IRES-Blast | Expresses bicistronic P2A construct of C-CRL4Cdt2 reporter(hCdt1(1-100)ΔCy-mCherry-) and APC/C reporter (hGeminin(1-110)) with blasticidin resistance |
| 193759 | pLV-mCherry-hCdt1(1-100)ΔCy | Expresses N-CRL4Cdt2 reporter(mCherry-hCdt1(1-100)ΔCy) |
| 193760 | pLV-TetOn-Cdt1-mCherry | Doxycyline-inducible (TetOn) human Cdt1 (C-terminal mCherry tag) |
| 193761 | pLV-TetOn-ND-Cdt1-mCherry | Doxycyline-inducible (TetOn) human Cdt1 (SV40 NLS localization and C-terminal mCherry tag) with mutations which abroggate its S phase degradation |
| 193762 | pLV-TetOn-NLS-mCherry | Doxycycline-inducible (TetOn) mCherry with SV40 NLS, control construct |
| 193763 | pCW-Cdt1-mCherry-Puro | Doxycyline-inducible (TetOn) human Cdt1 (C-terminal mCherry tag), expressed from all-in-one pCW vector (TetOn + rtTA) |
| 193764 | pCW-Cdt1ΔCy-mCherry-Puro | Doxycyline-inducible (TetOn) human Cdt1ΔCy (C-terminal mCherry tag), expressed from all-in-one pCW vector (TetOn + rtTA) |
| 193765 | pCW-Cdt1ΔPIP-mCherry-Puro | Doxycyline-inducible (TetOn) human Cdt1ΔPIP (C-terminal mCherry tag), expressed from all-in-one pCW vector (TetOn + rtTA) |
| 193766 | pCW-ND-Cdt1-mCherry-Puro | Doxycyline-inducible (TetOn) human Cdt1 (SV40 NLS localization and C-terminal mCherry tag) with mutations which abroggate its S phase degradation, expressed from all-in-one pCW vector (TetOn + rtTA) |
| 193767 | pCW-ND-Cdt1-HA-Puro | Doxycyline-inducible (TetOn) human Cdt1 (SV40 NLS localization and C-terminal HA tag) with mutations which abroggate its S phase degradation, expressed from all-in-one pCW vector (TetOn + rtTA) |
| 193768 | pCW-ND-Cdt1Δ499-546-HA-Puro | Doxycyline-inducible (TetOn) human Cdt1 (SDV40 NLS , C-terminal HA ) w/ mutations stopping S phase degradation and in MCM binding domain (Δ499-546), expressed from all-in-one pCW vector (TetOn + rtTA) |
| 193769 | pCW-ND-Cdt1ΔR198A/R210A-HA-Puro | Doxycyline-inducible (TetOn) human Cdt1 (SV40 NLS , C-terminal HA ) w/ mutations stopping S phase degradation and in MCM binding (R198A/R210A), expressed from all-in-one pCW vector (TetOn + rtTA) |
| 193770 | pCW-GemininΔDbox-HA-Puro | Doxycyline-inducible (TetOn) human Geminin (C-terminal HA tag) with Dbox removed to prevent degradation by APC/C, expressed from all-in-one pCW vector (TetOn + rtTA) |
| 193771 | pC1-ND-Cdt1-mCitrine | Human Cdt1 (SV40 NLS localization and C-terminal mCitrine tag) with mutations which abroggate its S phase degradation |