REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Human CREST sera/anti-centromere	Antibodies Inc.	#9101-02; RRID:AB_2687472
Mouse anti-CENP-A	GeneTex	#GTX13939; RRID:AB_369391
Rabbit anti-CENP-HK	Klare et al. (2015)	SI0930
Rabbit anti-CENP-TW	Klare et al. (2015)	SI0822
Donkey anti-human DyLight 405	Jackson Immuno Research	#709-475-149; RRID:AB_2340553
Goat anti-mouse Rhodamine Red	Jackson Immuno Research	#115-295-003; RRID:AB_2338756
Donkey anti-rabbit Alexa Fluor 647	Jackson Immuno Research	#711-605-152; RRID:AB_2492288
Rabbit anti-CyclinB1	Abcam	#ab32053; RRID:AB_731779
Mouse anti PCNA	Cell Signaling	#2586S; RRID:AB_2160343
Rat anti Tubulin	Abcam	#6160; RRID:AB_1140548
Donkey anti-mouse Alexa Fluor 647	Invitrogen	#6160; RRID:AB_1140548
Donkey anti-rabbit Rhodamine Red	Jackson Immuno Research	#711-295-152; RRID:AB_2340613
GFP-Booster Alexa Fluor 488	Chromotek	Gb2AF488; RRID:AB_2827573
Goat anti-human Alexa Fluor 647	Jackson Immuno Research	#109-605-003; RRID:AB_2337880
Bacterial and virus strains
E.coli: BL21CodonPlus(DE3)-RIL strain	Agilent Technologies	Cat#230280
S.frugiperda:Sf9 cells	ThermoFisher	Cat#12659017
Trichoplusia ni:BTI-Tnao38	Garry W Blissard Lab	N/A
Chemicals, peptides, and recombinant proteins
CENP-LN complex	Pentakota et al. (2017)	N/A
CENP-C(1–544)HKIM	This study	N/A
CENP-HKIM	This study	N/A
CENP-OPQUR complex	Pesenti et al. (2018)	N/A
CENP-TWSX complex	Walstein et al. (2021)	N/A
CENP-SX complex	Walstein et al. (2021)	N/A
CENP-TW complex	Walstein et al. (2021)	N/A
CENP-A nucleosomes (145 bp)	Walstein et al. (2021)	N/A
CENP-A nucleosomes (199 bp)	This study	N/A
Alexa Fluor 647C5 maleimide Protein labeling kit	ThermoFisher	Cat#A20347
Amylose resin	New England Biolabs	Cat#E8022
Streptavidin UltraLink Resin	Thermo Scientific	Cat#53114
400 mesh carbon-coated copper grids	Plano	G2400C
Uranyl Formate	SPI Supplies	Cas#16984-59-1
UltraAuFoil	QuantiFoil	N/A
Indole-3-acetic acid (IAA)	Merck	#I5148
DAPI	Sigma	Cat#D9542
Poly-D-Lysine	Sigma-Aldrich	Cat#A003E
Mowiol	Calbiochem	Cat#475904
Deposited data
human CCAN EM map and structure	This study	PDB: 7QOO and EMBD-14098
human CCAN-TW EM map	This study	EMDB-14099
Mendeley Dataset including uncropped microscopy images, SDS-PAGE scans, and spreadsheets of their quantification	This study	https://doi.org/10.17632/9smdtm4msj.1
Experimental models: Cell lines
Human DLD-1: CENP-CAID-EYFP/AID−EYFP	Fachinetti et al. (2015)	N/A
Human DLD-1: CENP-AEA/- CENP-CAmC/A	Hoffmann et al. (2020)	N/A
HeLa Flip-In T-Rex EGFP-CENP-M	Basilico et al. (2014)	N/A
Oligonucleotides
Primer GTGGACACTCTAAGGATCCGG	This study	N/A
Primer: Biotin-TEG- CCGGATCCTTAG AGTGTCCACCATC	This study	N/A
75bp-DNA: ATCCGTGGTAGAATAG 3GAAATATCTTCCTATAGAAACTAG ACAGAATGATTCTCAGAAACTCCT TTGTGATGGAT	This study	N/A
165bp-DNA: GTGGTAGAATAGGAAATATCTTCC TATAGAAACTAGACAGAATGATTC TCAGAAACTCCTTTGTGATGTGTG CGTTCAACTCACAGAGTTTAACCT TTCTTTTCATAGAGCAGTTAGGAA ACACTCTGTTTGTAATGTCTGCAA GTGGATATTCAGACGCCCTTG	This study	N/A
199bp-DNA: ATCGCCCTTGAGGCCTTCGTTGGAA ACGGGATTTCTTCATATTCTGCTAGA CAGAAGAATTCTCAGTAACTTCCTTG TGTTGTGTGTATTCAACTCACAGAGTT GAACGATCCTTTACACAGAGCAGACT TGAAACACTCTTTTTGTGGAATTTGCA GGCCTAGATTTCAGCCGCTTTGAGGT CAATCACCCCGTGGAT	This study	N/A
Recombinant DNA
pLIB	Addgene	Cat#80610
pLIB_6His-CENP-I	This study	N/A
pLIB_CENP-I	This study	N/A
pBiga	Addgene	CAT#80611
pBIGa_6His-CENP-C(1–544)_H_K_I_M	This study	N/A
pBIGa_6His-CENP-I_H_K_M	This study	N/A
pETDuet-1	Novagen	Cat#71146
pETDuet-1_6xHis-CENP-W_MBP-CENP-T	This study	N/A
pETDuet-1_6xHis-CENP-W_mScarlet-CENP-T	This study	N/A
pUC18	Addgene	Cat#50004
pUC18_199bp-DNA	This study	N/A
pUC18_165bp-DNA	This study	N/A
pUC18_75bp-DNA	This study	N/A
Software and algorithms
EPU	Thermo Fisher Scientific	N/A
TRanSHIRE	Stabrin et al. (2020)	https://transphire.readthedocs.io/en/latest/
MOTIONCORR 2.1	(Li et al., 2013)	http://cryoem.ucsf.edu/software/driftcorr.html
CTFFIND4	Rohou and Grigorieff (2015)	http://grigoriefflab.janelia.org/ctffind4
MOTIONCOR2	Zheng et al. (2017)	http://msg.ucsf.edu/em/software/motioncor2.html
SPHIRE	Moriya et al. (2017)	http://sphire.mpg.de
crYOLO	Wagner et al. (2019)	https://cryolo.readthedocs.io/en/stable/
RELION 3	Scheres Lab	https://www3.mrc-lmb.cam.ac.uk/relion/index.php?title = Main_PAGE
Chimera	Pettersen et al. (2004)	https://www.cgl.ucsf.edu/chimera/
ChimeraX	Pettersen et al. (2021)	https://www.cgl.ucsf.edu/chimerax/
Segger	Pintilie and Chiu (2012)	https://www.cgl.ucsf.edu/chimera/
Pymol	Schrödinger, LLC	https://pymol.org/2/
Namdinator	Kidmose et al., 2019	https://namdinator.au.dk/about/
COOT	Emsley et al. (2010)	https://www2.mrc-lmb.cam.ac.uk/personal/pemsley/coot/
PHENIX	Adams et al. (2010)	https://www.phenix-online.org
Molprobity	Chen et al. (2010)	https://www.phenix-online.org
DeepEMhancer	Sanchez-Garcia et al. (2021)	https://github.com/rsanchezgarc/deepEMhancer
DynDom6D	Veevers and Hayward (2019)	http://dyndom.cmp.uea.ac.uk/dyndom/dyndomDownload.jsp
GraphPad Prism Version 9.0.2 (134)	GraphPad Software Inc	http://www.graphpad.com
Fiji Version 2.0.0-rc-69/1.52n	(Schindelin et al., 2012)	http://ImageJ.nih.gov/ij/
softWoRx Version 7.0	GE Healthcare	N/A
CRaQ	Bodor et al. (2012)	N/A
Image Lab	Bio-rad	https://www.bio-rad.com/de-de/product/image-lab-software?ID=KRE6P5E8Z