REAGENT or RESOURCE	SOURCE	IDENTIFIER
Chemicals, peptides, and recombinant proteins
Certified Megabase Agarose	BioRad	Cat#161-3108
SYTOX green	Thermo Fischer	Cat# S7020
Alpha-Factor	Zymo Research	Cat#Y1001
RNAse A, DNase and protease-free	Thermo Fischer	Cat#EN0531
Concanavalin a from Canavalia ensiformis (Jack bean),Type VI, lyophilized powder	Sigma-Aldrich	Cat#L7647-250MG
Formaldehyde 37%	Sigma-Aldrich	Cat#F8775-4X25ML
Nocodazole	Sigma-Aldrich	Cat#M1404
DpnII	NEB	Cat#R0543M
Biotin-14-dCTP	Invitrogen	Cat#19518018
DNA Polymerase I Klenow Fragment	NEB	Cat#M0210L
T4 DNA ligase	Thermo Fischer	Cat#EL0014
Proteinase K	Thermo Scientific	Cat#EO0492
T4 DNA Polymerase	NEB	Cat#M0203L
Dynabeads MyOne Streptavidin C1	Invitrogen	Cat#65001
Hydroxyurea,98%, powder	Sigma-Aldrich	Cat# H8627-25G
Glass beads, acid-washed	Sigma-Aldrich	Cat#G8772-100G
Critical commercial assays
DNA size markers S. pombe chromosomal DNA	BioRad	Cat#170-3633
VK05 Lysing KIT	Ozyme	Cat#0042
NEBNext Ultra II FS kit	NEB	Cat#E7805L
Nextseq 500/550 High Output Kit v2.5 (150 Cycles)	Illumina	Cat# 20024907
NextSeq High-output 75-cycle V2.5 Kit	Illumina	Cat#20024906
Deposited data
Raw microscopy images	This study; Mendeley	https://data.mendeley.com/datasets/swdfkxzzk7/1
Raw FASQ files	This study	BioProject: PRJNA757122 https://www.ncbi.nlm.nih.gov/bioproject/PRJNA757122 Submission ID: SUB10236799
Experimental models: Organisms/strains
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0; chromosome# n = 16	Brachmann et al., 1998	BY4741
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0; chromosome# n = 3	Luo et al., 2018	JL381
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0; chromosome# n = 3	Luo et al., 2018	JL410
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0; chromosome# n = 2 size-matched size	Luo et al., 2018	JL402
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0; chromosome# n = 2 unmatched size	This study	JL498
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0 cdc15-2(ts); chromosome# n = 3	This study	LS381
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0 cdc15-2(ts); chromosome# n = 2 size-matched size	This study	LS402
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0 NUP49::mScarlet-S.p.HIS5; chromosome# n = 16	This study	LS125
S. cerevisiae. Genotype: MATahis3Δ1 leu2Δ0 met15Δ0 ura3Δ0 NUP49::mScarlet-S.p.HIS5; chromosome# n = 2 size-matched size	This study	LS126
Oligonucleotides
chr11R-gRNA for CRISPR-Cas9 AAATGAAGAAGTGCCATGGG	This study	n/a
chr5L-gRNA for CRISPR-Cas9 TTTCACATGCTCGACCGTTT	This study	n/a
cen7-gRNA TATTTTATTGTCGGTGTTTG	This study	n/a
Recombinant DNA
Plasmid: TEF1p-CRISPR-Cas9	Luo et al., 2018
Software and algorithms
ImageJ	Schneider et al., 2012	https://imagej.nih.gov/ij/
Matlab 2018	MATLAB 2018a, The MathWorks, Inc., Natick, Massachusetts, United States.	https://www.mathworks.com/products/matlab.html
		mat2pdb: https://fr.mathworks.com/matlabcentral/fileexchange/42957-read-and-write-pdb-files-using-matlab
FlowJo v10.0.7	Becton, Dickinson and Company; 2021	https://www.flowjo.com/
Bowtie 2	Langmead and Salzberg, 2012	http://bowtie-bio.sourceforge.net/bowtie2/index.shtml
Clustal Omega	Sievers et al., 2011	https://www.ebi.ac.uk/Tools/msa/clustalo/
PyMOL	Molecular Graphics System, Version 2.0 Schrödinger, LLC	https://pymol.org/edu/
DEseq2	Love et al., 2014	https://bioconductor.org/packages/release/bioc/html/DESeq2.html
Other
Saccharomyces Genome Database | SGD for S288c reference genome files		https://www.yeastgenome.org/
Yeast genome annotation pipeline - YGAP	Proux-Wera et al., 2012	http://wolfe.ucd.ie/annotation/index.html#questioncheck1