Key resources table.
| Reagent type (species) or resource |
Designation | Source or reference | Identifiers | Additional information |
|---|---|---|---|---|
| strain, strain background (Saccharomyces cerevisiae SK1) |
yDP71 | This paper | Cdc6-3V5 |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 CDC6-3V5::KANMX6/CDC6-3V5::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP120 | This paper | Orc6-3V5 |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 ORC6-3V5::KANMX6/ORC6-3V5::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP152 | This paper | cdk1-as, Cdc6-3V5 |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 cdc28-as1(F88G)/cdc28-as1(F88G) CDC6-3V5::KANMX6/CDC6-3V5::KANMX6 |
| strain, strain background (S. cerevisiae W303) |
yDP159 | This paper | Ime2 Purification |
W303 MATa bar1::hisG pep4::unmarked
LEU2::pGAL1,10-IME2(1–404)−3xFLAG |
| strain, strain background (S. cerevisiae SK1) |
yDP176 | This paper | ime2-as, Cdc6-3V5 |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 ime2-as1(M146G)/ime2-as1(M146G) CDC6-3V5::KANMX6/CDC6-3V5::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP177 | This paper | cdk1-as, ime2-as, Cdc6-3V5 |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 cdc28-as1(F88G)/cdc28-as1(F88G) ime2-as1(M146G)/ime2-as1(M146G) CDC6-3V5::KANMX6/CDC6-3V5::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP329 | This paper | Dpb11-3V5 |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 DPB11-3V5::KANMX6/DPB11-3V5::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP330 | This paper | Psf2-3V5 |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 PSF2-3V5::KANMX6/PSF2-3V5::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP335 | This paper | Cdc45-13myc |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 CDC45-13myc::KANMX6/CDC45-13myc::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP336 | This paper | Sld2-13myc |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 SLD2-13myc::KANMX6/SLD2-13myc::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP337 | This paper | Sld3-13myc |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 SLD3-13myc::KANMX6/SLD3-13myc::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP473 | This paper | Sld2-2TA-13myc |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 SLD2(T122A T143A)−13myc::KANMX6/ SLD2(T122A T143A)−13myc::KANMX6 |
| strain, strain background (S. cerevisiae W303) |
yDP554 | This paper | Ime2-AS Purification |
W303 MATa bar1::hisG pep4::unmarked
LEU2::pGAL1,10-IME2(1–404, M146G)−3xFLAG |
| strain, strain background (S. cerevisiae SK1) |
yDP642 | This paper | Sld2-2SA-13myc |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 SLD2(S128A S138A)−13myc::KANMX6/ SLD2(S128A S138A)−13myc::KANMX6 |
| strain, strain background (S. cerevisiae SK1) |
yDP644 | This paper | Sld2-4A-13myc |
SK1 MATa/alpha ura3::pGPD1-GAL4(848).
ER::URA3/ura3::pGPD1- GAL4(848).ER::URA3 GAL-NDT80::TRP1/GAL-NDT80::TRP1 SLD2(T122A S128A S138A T143A)−13myc:: KANMX6/SLD2(T122A S128A S138A T143A) −13myc::KANMX6 |
| strain, strain background (S. cerevisiae W303) |
ySK119 | Lõoke et al, 2017 (PMID: 28270517) |
Cdk1-Clb5 Purification | W303 MATa bar1::hisG pep4::unmarked URA3::pGAL1,10-Cdc28-His,Δ1–95-Clb5-Flag |
| strain, strain background (S. cerevisiae SK1) |
yST135 | This paper | Mcm2-7 Purification |
W303 MATa bar1::hisG pep4::unmarked
TRP1::pSKM003(pGAL1,10-MCM6,MCM7) HIS3::pSKM004-(pGAL1,10-MCM2,Flag-MCM3) LYS2::pSKM002-(pGAL1,10-MCM4,MCM5) |
| strain, strain background (S. cerevisiae W303) |
yST144 |
Ticau et al. (2015)
(PMID: 25892223) |
Mcm2-7-Cdt1 Purification |
W303 MATa bar1::hisG pep4::unmarked
TRP1::pSKM003(pGAL1,10-MCM6,MCM7) HIS3::pSKM004-(pGAL1,10-MCM2,Flag-MCM3) LYS2::pSKM002-(pGAL1,10-MCM4,MCM5) URA3::pALS1(pGAL1,10-Cdt1,GAL4) |
| strain, strain background (S. cerevisiae W303) |
A4370 | Angelika Amon | cdk1-as | W303 MATa bar1::hisG cdc28-as1(F88G) |
| strain, strain background (S. cerevisiae W303) |
ySDORC | John Diffley | ORC purification |
W303 MATa bar1::hyg pep4::kanMX
TRP1::pGAL1,10-ORC5,ORC6 HIS3::pGAL1,10-ORC3,ORC4 URA3::pGAL1, 10-CBP-TEV-ORC1,ORC2 |
| antibody | poly ORC (Orc1 and Orc2 western blots and ORC ChIP) |
HM1108 (Bell Lab) | ||
| antibody | Cdt1 | HM5353 (Bell Lab) | ||
| antibody | poly MCM (Mcm3 and Mcm6 western blots) |
UM174 (Bell Lab) | ||
| antibody | poly MCM (Mcm2-7 ChIP) |
UM185 (Bell Lab) | ||
| antibody | Mcm2 | Santa Cruz, yN-19 (code sc-6680) |
RRID:AB_648843 | |
| antibody | Mcm7 | Santa Cruz, yN-19 (code SC-6688) |
RRID:AB_647936 | |
| antibody | PGK1 | Invitrogen (catalog #459250) |
RRID:AB_2532235 | |
| recombinant DNA reagent (plasmid) |
pSKM033 |
Kang et al. (2014)
(PMID: 25087876) |
Cdc6 purification | pGEX-GST-3C-FLAG-CDC6 |
| recombinant DNA reagent (plasmid) |
pALS16 | This study | Cdt1 purification | pGEX-GST-3C-CDT1 |
| chemical compound, drug | 1-NM-PP1 | Toronto Research Chemicals (catalog #A603003) |
||
| chemical compound, drug | 1-NA-PP1 | Cayman Chemical Co., (catalog #NC1049860) |