Extended Data Table 1 |. Characterization of cysteines in truncated dynein.

Cysteines of the dynein motor domain were categorized into three groups based on available cryo-EM structures of yeast dynein (PDB ID: 4AKG)²³. The surface-exposed cysteines were accessible to surrounding water over 5 Å, partially exposed cysteines were solvent accessible to over 2.5 Å, and buried cysteines to less than 2.5 Å. Surface-exposed cysteines of the GST-dimerization were also removed. SpyCatcher and SpyTag do not contain any cysteines and are therefore omitted from this table.

Protein	Surface exposed	Partially exposed	Buried
GFP		C48	C70
GST	C84 C137 C177	C168
Dyn314kDa	C2037 C2268 C2372 C3499 C4016	C1405 C1663 C1718 C2210 C2220 C2724 C2892 C3141 C3206 C3324 C3684 C3704 C3959	C1428 C1626 C1793 C1822 C1846 C1960 C1980 C2057 C2078 C2417 C2486 C2535 C2603 C2806 C2814 C2912 C3382 C3652 C3877 C3928 C4011
Total	8	15	22