REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Sheep polyclonal anti-Psf2	Gambus lab	Gambus et al.56
Sheep polyclonal anti-Cdc45	Gambus lab	Gambus et al.56
Sheep polyclonal anti-GINS	Gambus lab	Tarcan et al.57
Rabbit polyclonal anti-DONSON	Gambus lab	Kingsley et al.42
Sheep polyclonal anti-DONSON	Gambus lab	Kingsley et al.42
Sheep polyclonal anti-Mcm7	Gambus lab	Priego Moreno et al.58
Sheep polyclonal anti-RecQ4	Gambus lab	Moreno et al.59
Sheep polyclonal anti-TopBP1	Gambus lab	Moreno et al.59
Sheep polyclonal anti-Treslin	Blow lab	Volpi et al.60
Sheep polyclonal anti-MTBP	Blow lab	Volpi et al.60
IgG from rabbit serum	Sigma-Aldrich	RRID: AB_1163659
Polyclonal rabbit anti-FLAG	Sigma-Aldrich	RRID: AB_439687
Monoclonal mouse anti-Mcm2	BD Transduction labs	RRID: AB_2141952
Monoclonal mouse anti-PCNA	Santa Cruz Biotechnology	RRID: AB_628110
Monoclonal mouse anti-PCNA	Sigma Aldrich	RRID: AB_477413
Bacterial and virus strains
Rosetta™(DE3) pLysS Competent Cells	Novagen (Merck)	70956
BL21(DE3) Competent Cells	Novagen (Merck)	69451
Chemicals, peptides, and recombinant proteins
Aphidicolin	Bio-Techne Limited	Cat# 5736/1
Caffeine	Sigma-Aldrich	CAS: 58-08-2
ANTI-FLAG® M2 Magnetic Beads	Sigma-Aldrich	Cat# M8823
3X FLAG Peptide	Stratech Scientific Ltd	A6001
HisTrap HP	Cytiva	Cat# 17524801
HiTrap Heparin HP	Cytiva	Cat# 17040701
Cdc45-TEV-His10-FLAG5	This study	Addgene agreement 83268
DONSON WT	Gambus lab	Kingsley et al.42
DONSON DEW (D374A; E377A; W381A)	This study	Addgene agreement 83268
DONSON S437A	This study	Addgene agreement 83268
DONSON WM (W234L; M463T)	This study	Addgene agreement 83268
DONSON 3A (V473A; R476A; Y481A)	This study	Addgene agreement 83268
DONSON D24 (D1-24)	This study	Addgene agreement 83268
DONSON DEWS (D374A; E377A; W381A; S437A)	This study	Addgene agreement 83268
DONSON DD1 (D1-155)	Gambus lab	Kingsley et al.42
GINS: (Psf1, Psf2, Psf3 and Sld5)	Gambus lab	Tarcan et al.57
Graphene oxide	Sigma-Aldrich	763705-25ML
Deposited data
Xenla dCMGDo consensus cryo-EM map	This study	EMDB: EMD-18191
Xenla dCMGDo ATPase cryo-EM map	This study	EMDB: EMD-18192
Xenla sCMG consensus cryo-EM map	This study	EMDB: EMD-18195
Xenla dCMGDo – without MCM ATPase PDB coordinates	This study	PDB: 8Q6O
Xenla dCMGDo – MCM ATPase PDB coordinates	This study	PDB: 8Q6P
Xenopus laevis Cdc45-FLAG interactions on chromatin during DNA replication – Mass spectrometry data project	This study	PRIDE: PXD044422
Experimental models: Cell lines
293FT	Thermo Fisher Scientific	R70007
Experimental models: Organisms/strains
Xenopus laevis	Gambus lab	N/A
Oligonucleotides
DONSON DEW (D374A; E377A; W381A) forward: CTCTGCGCTAGAAGAGATGGGAGTTGAAGA TAAAATTAAAAAG;	Sigma-Aldrich	N/A
DONSON DEW (D374A; E377A; W381A) reverse: AAGCTCGCATCTTCCGCGCTTTCATC CGATGCCTC	Sigma-Aldrich	N/A
DONSON S437A forward: AAACTGTAAGGC GATTGTCGCTGC	Sigma-Aldrich	N/A
DONSON S437A reverse: TGAGGAAGTT CAGTAAAG	Sigma-Aldrich	N/A
DONSON W234L forward: GTCGCTTCCTCT GGTACAGCTTTTTC	Sigma-Aldrich	N/A
DONSON W234L reverse: GGATGAATCCAG TGCACAAG	Sigma-Aldrich	N/A
DONSON M463T forward: AGGAGCAACAA CGCACGCGCTTA	Sigma-Aldrich	N/A
DONSON M463T reverse: CGGAATGCGA CTGGAGAC	Sigma-Aldrich	N/A
DONSON 3A (V473A; R476A; Y481A) forward: GAACTCTGGTGCGAAGGATCAGTTTAGTCTGG;	Sigma-Aldrich	N/A
DONSON 3A (V473A; R476A; Y481A) reverse: ACCGCAGTCTTCGCATTCACACTCCTTG CCTTAAG	Sigma-Aldrich	N/A
DONSON D24 (Δ1-24) forward: AGGAGCAGG AGCGAGGCT	Sigma-Aldrich	N/A
DONSON D24 (Δ1-24) reverse: CAGATCCTCT TCTGAGATGAGTTTTTGTTCG	Sigma-Aldrich	N/A
DONSON DD1 (Δ1-155) forward: TCCGTGAC TTTTCCTGCT	Sigma-Aldrich	N/A
DONSON DD1 (Δ1-155) reverse: CATTTATTT CCAGGTGTACC	Sigma-Aldrich	N/A
Recombinant DNA
pET28a-DONSON	Gambus lab	Addgene: 208797
pET28a-DONSON ΔD1	Gambus lab	Addgene: 208804
pET28a-DONSON-WM (W234L, M463T)	This study	Addgene: 208798
pET28a-DONSON-DEWS (D374A, E377A, W381A, S437A)	This study	Addgene: 208799
pET28a-DONSON-3A (V473A, R476A, Y481A)	This study	Addgene: 208800
pET28a-DONSON Δ24 (1-24 removed)	This study	Addgene: 208801
pET28a-DONSON-S437A	This study	Addgene: 208803
pET28a-DONSON-DEW (D374A, E377A, W381A)	This study	Addgene: 208802
Software and algorithms
crYOLO 1.4.0	Wagner et al.61	https://cryolo.readthedocs.io/en/stable/
Relion 3.0.4 and 4.0.0	Kimanius et al.62; Zivanov et al63	https://www.mrc-lmb.cam.ac.Uk/relion/index.php?title=Main_Page
cryoSPARC v4.2.1	Punjani et al.64	https://cryosparc.com/
Gctf	Zhang et al.65	https://sbgrid.org/software/titles/gctf
CtfFind-4.1	Rohou et al.66	https://grigoriefflab.umassmed.edu/ctffind4
Topaz-0.2.5	Bepler et al.67	https://cb.csail.mit.edu/cb/topaz/
Coot V0.9.8.1	Emsley et al.68	https://www.mrc-lmb.cam.ac.uk/personal/pemsley/coot/
PHENIX V1.20.1	Liebschner et al.69	http://www.phenix-online.org/
UCSF Chimera V1.16	Pettersen et al.70	https://www.cgl.ucsf.edu/chimera/
UCSF ChimeraX	Goddard et al.71	https://www.rbvi.ucsf.edu/chimerax/
Mascot v 2.8.0.1	Matrix Science, London, UK	https://www.matrixscience.com/
Scaffold v 5.1.0	Proteome Software Inc., Portland	https://proteomesoftware.com/
AcquireMP v 2.5	Refeyn Ltd., Oxford, UK	https://www.refeyn.com/
DiscoverMP v 2.5	Refeyn Ltd., Oxford, UK	https://www.refeyn.com/
GraphPad Prism v 10.0.2	GraphPad Software LLC	https://www.graphpad.com/