The HIV-1 IN CCD (residues 50–212) containing the F185H mutation was expressed and purified as described [23]. The protein was concentrated to 8 mg/ml and crystallized using hanging-drop vapor diffusion method with a crystallization buffer consisting of 100 mM sodium cacodylate pH 6.5, 100 mM ammonium sulfate, 10% (w/v) PEG 8000, and 5 mM DTT. Crystallization drops were prepared using an equal volume of protein and well solution. Crystallization trays were prepared on ice at room temperature and then transferred to 4°C for storage. Crystals formed within one week to a month. Crystals were transferred to a drop containing crystallization solution, 5 mM of STP0404, and 10% DMSO. Crystals were soaked overnight prior to data collection. Crystal data were collected on a Rigaku Micromax-007 at 100 K. Data were integrated and scaled using HKL3000 [37] and Scalepack [38]. Phaser [39] in the PHENIX suite [40] was used to run molecular replacement using Protein Data Bank code 4O55 as a search model [23]. Phenix.refine [41] was used for data refinement, and manual refinement was done in Coot [42]. The coordinates are deposited in the Protein Data Bank under accession codes 7KE0. The data and refinement statistics are given in S1 Table.

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