REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial and Virus Strains
Escherichia coli: Rosetta™ 2(DE3) strain: F–ompT hsdSB(rB– mB–) gal dcm (DE3) pRARE2 (CamR)	Novagen / Merck Millipore	Cat# 71400
Chemicals, Peptides, and Recombinant Proteins
3X FLAG peptide	Sigma	Cat# F4799
Anti-FLAG M2 affinity gel	Sigma	Cat# A2220
Calmodulin-Sepharose 4B	GE Healthcare / Sigma	Cat# 17-0529-01
Calmodulin Affinity Resin	Agilent	Cat# 214303
Bio-Gel HT Hydroxyapatite	Bio-Rad	Cat# 1300150
Cdt1-Mcm2-7	Coster et al., 2014	N/A
ORC	Frigola et al., 2013	N/A
Cdc6	Frigola et al., 2013	N/A
DDK	On et al., 2014	N/A
Sld3/7	Yeeles et al., 2015	N/A
Cdc45	Yeeles et al., 2015	N/A
Dpb11	Yeeles et al., 2015	N/A
Sld2	Yeeles et al., 2015	N/A
GINS	Yeeles et al., 2015	N/A
Pol ε	Yeeles et al., 2015	N/A
S-CDK	Yeeles et al., 2015	N/A
Mcm10	Yeeles et al., 2015	N/A
Pol α	Yeeles et al., 2015	N/A
RPA	Devbhandari et al., 2017	N/A
Ctf4	Yeeles et al., 2015	N/A
Topo I	Yeeles et al., 2017	N/A
Mrc1	Yeeles et al., 2017	N/A
Csm3/Tof1	Yeeles et al., 2017	N/A
RFC	Yeeles et al., 2017	N/A
PCNA	Yeeles et al., 2017	N/A
Pol δ	Yeeles et al., 2017	N/A
Histones	Kurat et al., 2017	N/A
Nap1	Kurat et al., 2017	N/A
Isw1a	Kurat et al., 2017	N/A
FACT	Kurat et al., 2017	N/A
Nhp6	Kurat et al., 2017	N/A
Experimental Models: Organisms/Strains
yAM33 (Cdt1-Mcm2-7 purification)	Coster et al., 2014	N/A
ySD-ORC (ORC purification)	Frigola et al., 2013	N/A
ySDK8 (DDK purification)	On et al., 2014	N/A
yTD6 (Sld3/7 purification)	Yeeles et al., 2015	N/A
yTD8 (Sld2 purification)	Yeeles et al., 2015	N/A
yJY13 (Cdc45 purification)	Yeeles et al., 2015	N/A
yJY26 (Dpb11 purification)	Yeeles et al., 2015	N/A
yAJ2 (Pol ε purification)	Yeeles et al., 2015	N/A
yAE37 (S-CDK purification)	Yeeles et al., 2015	N/A
yAE40 (Ctf4 purification)	Yeeles et al., 2015	N/A
yJY23 (Pol α purification)	Yeeles et al., 2015	N/A
yAE34 (Pol δ purification)	Yeeles et al., 2017	N/A
yAE41 (RFC purification)	Yeeles et al., 2017	N/A
yAE42 (Topo I purification)	Yeeles et al., 2017	N/A
yAE48 (Csm3/Tof1 purification)	Yeeles et al., 2017	N/A
yJY32 (Mrc1 purification)	Yeeles et al., 2017	N/A
yCFK1 (Isw1a purification)	Kurat et al., 2017	N/A
Oligonucleotides
See Table S1 for all DNA oligonucleotides used in this study
Recombinant DNA
pBluescript II KS(–) Phagemid	Agilent Technologies	Cat# 212208-51
ZN3: replication template	This paper	N/A
ZN5SP1: replication template	This paper	N/A
M13mp18 Single-stranded DNA: primase assay	New England Biolabs	Cat# N4040S
λ DNA-HindIII Digest: molecular weight markers	New England Biolabs	Cat# N3012S
pJM126 (RPA purification)	Addgene	#49339
pAM3 (Cdc6 purification)	Frigola et al., 2013	N/A
vJY19 (PCNA purification)	Yeeles et al., 2017	N/A
pRJ1228-Nhp6 (Nhp6 purification)	Ruone et al., 2003	N/A
pCDFduet.H2A-H2B (Histones purification)	Kingston et al., 2011	N/A
pETduet.H3-H4 (Histones purification)	Kingston et al., 2011	N/A
pJFDJ5 (GINS purification)	Yeeles et al., 2015	N/A
pET28a-Mcm10 (Mcm10 purification)	Yeeles et al., 2015	N/A
pTF175 (FACT purification)	Biswas et al., 2005	N/A
pJW22 (FACT purification)	Biswas et al., 2005	N/A
pCFK1 (Nap1 purification)	Kurat et al., 2017	N/A
Software and Algorithms
ImageJ	National Institute of Health	https://imagej.nih.gov/ij/