REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Rabbit polyclonal anti-SMARCAD1	Bethyl	A301-593A; RRID: AB_1078836
Rabbit polyclonal anti-KAP1	Abcam	Ab10483; RRID: AB_297222
Mouse monoclonal anti-α-Tubulin	Francis Crick Institute	TAT-1; RRID: AB_10013740
Mouse monoclonal anti-HA	Francis Crick Institute	12CA5; RRID: AB_2532070
Rabbit polyclonal anti-FLAG	Sigma	F7425; RRID: AB_439687
Sheep anti-mouse IgG HRP-linked F(ab’)2 fragments	GE Healthcare	NA9310; RRID: AB_772193
Donkey anti-rabbit IgG HRP-linked F(ab’)2 fragments	GE Healthcare	NA9340; RRID: AB_772191
Bacterial and Virus Strains
E. coli BL21-CodonPlus® (DE3)-RIL	Agilent	#230245
E. coli BL21-CodonPlus® (DE3)-RP	Agilent	#230255
E. coli BL21(DE3) Rosetta™ 2	Novagen	71400
E. coli BL21(DE3)-R3-pRARE2	Structural Genomics Consortium, Oxford	N/A
Chemicals, Peptides, and Recombinant Proteins
FLAG-SMARCAD1 (WT)	This Study	N/A
FLAG-SMARCAD1 CUE1mt	This Study	N/A
FLAG-SMARCAD1 CUE2mt	This Study	N/A
FLAG-SMARCAD1 CUE1mt,2mt	This Study	N/A
FLAG-CUE1,2 (WT)	This Study	N/A
FLAG-CUE1mt,2	This Study	N/A
FLAG-CUE1,2mt	This Study	N/A
FLAG-CUE1mt,2mt	This Study	N/A
FLAG-CUE1,2 K161A	This Study	N/A
FLAG-CUE1,2 Q163A	This Study	N/A
FLAG-CUE1,2 T164A	This Study	N/A
FLAG-CUE1,2 K166A	This Study	N/A
FLAG-CUE1,2 E167A	This Study	N/A
FLAG-CUE1,2 L168A	This Study	N/A
FLAG-CUE1,2 F169A	This Study	N/A
FLAG-CUE1,2 P170A	This Study	N/A
FLAG-CUE1,2 Q171A	This Study	N/A
FLAG-CUE1,2 D188A	This Study	N/A
FLAG-CUE1,2 A192G	This Study	N/A
FLAG-CUE1,2 L195A	This Study	N/A
FLAG-CUE1,2 L196A	This Study	N/A
FLAG-CUE1,2 Δ200-230	This Study	N/A
FLAG-CUE1	This Study	N/A
FLAG-CUE1mt	This Study	N/A
HA-KAP1	This Study	N/A
KAP1 G53-K434	This Study	N/A
KAP1 G53-K434 ΔBBX1 (Δ141-202)	This Study	N/A
KAP1 S33-K434	This Study	N/A
KAP1 S33-K434 M335A	This Study	N/A
KAP1 S33-K434 I338A	This Study	N/A
KAP1 S33-K434 E342A	This Study	N/A
KAP1 S33-K434 R347A	This Study	N/A
KAP1 S33-K434 L376A	This Study	N/A
KAP1 S33-K434 M378A	This Study	N/A
KAP1 S33-K434 I379A	This Study	N/A
KAP1 S33-K434 V380A	This Study	N/A
Ubiquitin	Boston Biochem	U-100H
Ulp1	Purified as per Peter Cherepanov, FCI	N/A
Lysozyme	Sigma	L6876
Micrococcal nuclease	NEB	M0247
Ni-NTA agarose	Qiagen	30250
HisTrap HP	GE Healthcare	17524801
HiTrap Heparin HP column	GE Healthcare	17040701
ProSwift WCX-1S	Thermo Fisher Scientific	064295
ProSwift SAX-1S	Thermo Fisher Scientific	064293
MAbPac SEC-1 column	Thermo Fisher Scientific	074696
Mono Q™ 5/50 GL column	GE Healthcare	17-5166-01
HiPrep 16/60 Sephacryl S-400 HR	GE Healthcare	28-9356-04
Mono Q™ 10/100 GL	GE Healthcare	17-5167-01
HiLoad 16/600 Superdex 200 column	GE Healthcare	28-9893-35
Amicon Ultra-15 30K MWCO spin concentrator	Millipore	UFC903008
Amicon Ultra-4 10K MWCO spin concentrators	Millipore	UFC801024
Microcon-30kDa Centrifugal Filter Unit	Millipore	MRCF0R030
anti-HA (3F10) affinity matrix	Roche	11 815 016 001; RRID: AB_390914
HA peptide	Peptide Chemistry STP, FCI	N/A
anti-FLAG M2 affinity gel	Sigma	A2220; RRID: AB_10063035
FLAG peptide	Peptide Chemistry STP, FCI	N/A
Bio-Rad Protein Assay	Bio-Rad	5000006
InstantBlue™	Expedeon	ISB1L
SilverQuest Silver Staining Kit	Invitrogen	LC6070
SYPRO® Ruby	Thermo Fisher Scientific	S11791
Deposited Data
TRIM25 coiled-coil domain X-ray structure	PDB	PDB 4CFG
TRIM28 B-box 2 domain X-ray structure	PDB	PDB 2YVR
TRIM56 RING domain X-ray structure	PDB (Fiskin et al., 2017)	PDB 5JW7
GP78 CUE domain NMR structure	PDB (Liu et al., 2012)	PDB 2LVN
KAP1 RBCC-SMARCAD1 CUE1,2 X-ray structure	This Study	PDB 6H3A
KAP1 RBCC ΔBBX1-SMARCAD1 CUE1,2 X-ray structure	This Study	PDB 6QU1
Experimental Models: Cell Lines
T-Rex-293 cell line	Thermo Fisher Scientific	R71007
293 T-Rex F-SMARCAD1 716R	This Study	N/A
Experimental Models: Organisms/Strains
SMARCAD1 cDNA	Human	UniProt Q9H4L7
KAP1 cDNA	Human	UniProt Q13263
Recombinant DNA
pcDNA4/TO vector	Thermo Fisher Scientific	V102020
pSMARCAD1-716R-CUE1mt,2mt/TO	This Study	N/A
pGIPZ-SMARCAD1 V2LHS-51716	Dharmacon	V2LHS-51716
pGEX-6P-1	GE Healthcare	28-9546-48
pGEX6P1-KAP1	This Study	N/A
pGEX6P1-KAP1 S33-K434	This Study	N/A
pGEX6P1-KAP1 D202-K434	This Study	N/A
pGEX6P1-KAP1 L592-P835	This Study	N/A
pET28a-SUMO	Peter Cherepanov, FCI	N/A
pET28a-SUMO-HA-KAP1	This Study	N/A
pET28a-SUMO-KAP1 S33-K434	This Study	N/A
pET28a-SUMO-KAP1 S33-K434 M335A	GenScript, This Study	N/A
pET28a-SUMO-KAP1 S33-K434 I338A	GenScript, This Study	N/A
pET28a-SUMO-KAP1 S33-K434 E342A	GenScript, This Study	N/A
pET28a-SUMO-KAP1 S33-K434 R347A	GenScript, This Study	N/A
pET28a-SUMO-KAP1 S33-K434 L376A	GenScript, This Study	N/A
pET28a-SUMO-KAP1 S33-K434 M378A	GenScript, This Study	N/A
pET28a-SUMO-KAP1 S33-K434 I379A	GenScript, This Study	N/A
pET28a-SUMO-KAP1 S33-K434 V380A	GenScript, This Study	N/A
pET28a-SUMO-F-SMARCAD1	This Study	N/A
pET28a-SUMO-F-SMARCAD1-CUE1mt,2	This Study	N/A
pET28a-SUMO-F-SMARCAD1-CUE1,2mt	This Study	N/A
pET28a-SUMO-F-SMARCAD1-CUE1mt,2mt	This Study	N/A
pET28a-SUMO-F-CUE1,2 (S95-N347)	This Study	N/A
pET28a-SUMO-F-CUE1mt,2	This Study	N/A
pET28a-SUMO-F-CUE1,2mt	This Study	N/A
pET28a-SUMO-F-CUE1mt,2mt	This Study	N/A
pET28a-SUMO-F-CUE1,2 K161A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 Q163A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 T164A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 K166A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 E167A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 L168A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 F169A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 P170A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 Q171A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 D188A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 A192G	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 L195A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 L196A	GenScript, This Study	N/A
pET28a-SUMO-F-CUE1,2 Δ200-230	GenScript, This Study	N/A
pET28a-SUMO-CUE1,2	This Study	N/A
pET28a-SUMO-F-CUE1 (S95-E237)	This Study	N/A
pET28a-SUMO-F-CUE1mt	This Study	N/A
pCDF-Duet1-SUMO	Peter Cherepanov, FCI	N/A
pCDF-SUMO-KAP1 S33-K434	This Study	N/A
Software and Algorithms
Origin	Malvern	N/A
DIALS 1.11	DIALS	N/A
Phenix autosol	Phenix	N/A
Phenix refine	Phenix	N/A
PHASER	CCP4	N/A
UCSF Chimera	UCSF	N/A
Other
Diamond Light Source beamline i03	N/A	N/A
Diamond Light Source beamline I04-1	N/A	N/A