REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
rat anti-HA (3F10)	Roche	Cat# 3F10; RRID:AB_2314622
rabbit anti-hCENP-C	Black lab	N/A
mouse anti-hCENP-A	Abcam	Cat# ab13939; RRID:AB_300766
rabbit anti-dCENP-C	Lehner lab	N/A
rabbit anti-V5	Sigma-Aldrich	Cat# V8137; RRID:AB_261889
mouse anti-V5	Thermo Fisher	Cat# R960-25; RRID:AB_2556564
rabbit anti-Myc	Abcam	Cat# ab9106; RRID:AB_307014
chicken anti-dCENP-A	This paper	N/A
mouse anti-mCherry	Sacchani lab	N/A
mouse anti-GFP	Sacchani lab	N/A
mouse anti-HA 12CA5	Sigma-Aldrich	Cat#11583816001; RRID:AB_514505
rabbit anti-HA	Abcam	Cat# ab9110, RRID:AB_307019
Bacterial and Virus Strains
ElectroMAX DH5α-E Competent Cells	ThermoFisher Scientific	11319019
Chemicals, Peptides, and Recombinant Proteins
dCENP-A(101-225)	This paper	pET3a
dH4	Karolin Luger	pET3a
Human H4	This paper	pEC-K-3C-His-GST
CAL1(1-160)	This paper	pEC-K-3C-His-GST
CAL1 (841-979)	This paper	pEC-K-3C-His-GST
dCENP-C (1264-1411)	This paper	pEC-K-3C-His-GST
Experimental Models: Cell Lines
U2OS-LacO (female)	S. Janicki lab
U2OS-LacO- CAL1V5_T2A_ dCENP-A_T2A_ HAdCENP-C (clone 17)	This paper	N/A
U2OS-LacO-dCENP-A (clone4)	This paper	N/A
Schneider S2 cells (L2-4; male)	Botchan lab	N/A
Oligonucleotides
αSatCh17-f2: CGTTGGAAACGGGATAATTTCAGCTGACTA	This paper	N/A
αSatCh17-r2: CACAGAGTGGTCCAAATATCCACTTGTAGA	This paper	N/A
αSatCh21-f2: GGCCTTTCATAGAGCAGGTTTGAAACACTC	This paper	N/A
αSatCh21-r2: CAAATATCCACTTGCAGATTCCACAAAAAG	This paper	N/A
LacO_F TAGAGGCGCCGAATTCCA	Mendiburo et al., 2011	N/A
LacO_R2 ATCCGCTCACAATTCCAC	Mendiburo et al., 2011	N/A
Recombinant DNA
pDS47_dCENP-A_HA	This paper	N/A
pDS47_H3_HA	This paper	N/A
pDS47_H3L1α2_HA	This paper	N/A
pDS47_dCENP-AC_HA	This paper	N/A
pDS47_dCENP-A α2+C_HA	This paper	N/A
pDS47_H3 L1α2+C _HA	This paper	N/A
pDS47_H3 α2+C _HA	This paper	N/A
pDS47_H3 C _HA	This paper	N/A
pDS47_dCENP-A L1α2 _HA	This paper	N/A
pDS47_dCENP-A L1α2+C _HA	This paper	N/A
pDS47_H3 L1+C _HA	This paper	N/A
pDS47_H3 L1α2+C(223) _HA	This paper	N/A
pDS47_H3 L1α2+C(222) _HA	This paper	N/A
pDS47_H3 L1α2+C(219) _HA	This paper	N/A
pDS47_H3 L1α2+C(218) _HA	This paper	N/A
pDS47_H3 L1α2+C(206) _HA	This paper	N/A
pMT_CAL1_HA	This paper	N/A
pMT_CAL1(NT)_HA	This paper	N/A
pMT_CAL1(CT)_HA	This paper	N/A
pN2_CMV_dCENP-A_GFPLacI	This paper	N/A
pN2_CMV _H3_ GFPLacI	This paper	N/A
pN2_CMV _H3L1α2_ GFPLacI	This paper	N/A
pN2_CMV _dCENP-AC_ GFPLacI	This paper	N/A
pN2_CMV _dCENP-A α2+C_ GFPLacI	This paper	N/A
pN2_CMV _H3 L1α2+C _ GFPLacI	This paper	N/A
pN2_CMV _H3 α2+C _ GFPLacI	This paper	N/A
pN2_CMV _H3 C _ GFPLacI	This paper	N/A
pN2_CMV _dCENP-A L1α2 _ GFPLacI	This paper	N/A
pN2_CMV _dCENP-A L1α2+C _ GFPLacI	This paper	N/A
pN2_CMV _H3 L1+C _ GFPLacI	This paper	N/A
pN2_CMV _ GFPLacI_dCENP-C(CT)	This paper	N/A
pN2_CMV _ GFPLacI	This paper	N/A
pN2_CMV _ GFPLacI_HJURP	This paper	N/A
pN2_CMV_CAL1_GFPLacI	This paper	N/A
pN2_CMV _ GFPLacI_dCENP-C	This paper	N/A
pN2_CMV_CAL1_V5	This paper	N/A
pN2_CMV_dCENP-A_V5	This paper	N/A
pN2_CMV_tetR_V5	This paper	N/A
pN2_CMV _dCENP-AC_V5	This paper	N/A
pN2_CMV _dCENP-AC_V5	This paper	N/A
pN2_CMV _ HA_dCENP-C	This paper	N/A
pN2_CMV_CAL1_HA	This paper	N/A
pN2_CMV_dCENP-A_HA	This paper	N/A
pN2_CMV _H3_ HA	This paper	N/A
pN2_CMV_CAL1(NT)_HA	This paper	N/A
pN2_CMV_CAL1(CT)_HA	This paper	N/A
pN2_CMV_CAL1(MID)_HA	This paper	N/A
pN2_CMV_CAL1(1-160)_HA	This paper	N/A
pN2_CMV_CAL1(101-407)_HA	This paper	N/A
pN2_CMV _ HA_dCENP-C(NT)	This paper	N/A
pN2_CMV _ HA_dCENP-C(CT)	This paper	N/A
pN2_CMV _ HA_dCENP-C(MID)	This paper	N/A
pN2_CMV _ HA_dCENP-C(1009-1263)	This paper	N/A
pN2_CMV _ HA_dCENP-C(1264-1411)	This paper	N/A
pN2_CMV _ HA_dCENP-C(1264-1411)L1357E-L1406E	This paper	N/A
pN2_CMV _ HA_dCENP-C_L1357E-L1406E	This paper	N/A
pN2_CMV_HA_hCENP-C	This paper	N/A
pN2_CMV_hCENP-A_HA	This paper	N/A
pcDNA3_CMV_HA_HJURP	This paper	N/A
pN2_CMV_CAL1(NT)_ GFPLacIw	This paper	N/A
pN2_CMV_CAL1(1-160)_ GFPLacIw	This paper	N/A
pN2_CMV _ GFPLacIw	This paper	N/A
pN2_CMV_dCENP-A(Y187A-L188Q)_GFPLacIw	This paper	N/A
pN2_CMV _dCENP-AC_ GFPLacIw	This paper	N/A
pN2_CMV_dCENP-A_GFPLacIw	This paper	N/A
pN2_CMV_CAL1_ GFPLacIw	This paper	N/A
pN2_CMV_ GFPLacIw_dCENP-C	This paper	N/A
pN2_CMV_tetR_Myc	This paper	N/A
pN2_CMV_dCENP-A_SNAP	This paper	N/A
pN2_CMV_dCENP-A_mCherry	This paper	N/A
pN2_CMV_CAL1V5_T2A_ dCENP-A_T2A_ HAdCENP-C	This paper	N/A
pET3a_dCENP-A(101-225)	This paper	N/A
pET3a_dH4	Karolin Luger	N/A
pEC-K-3C-His-GST_hH4	This paper	N/A
pEC-K-3C-His-GST_CAL1(1-160)	This paper	N/A
pEC-K-3C-His-GST_CAL1 (841-979)	This paper	N/A
pEC-K-3C-His-GST_dCENP-C (1264-1411)	This paper	N/A
Software and Algorithms
ImageJ
Prism	GraphPad	Prism 8.0
Other
Adobe Illustrator	Adobe	CC 2019
Adobe Photoshop	Adobe	CC 2019
softWoRx Explorer Suite 2.0	Applied Precision	2.0