The three sets of starting coordinates were generated from the microsecond trajectory of T4 lysozyme bound to p-xylene, with the first being the input coordinates (T0), the second being a snapshot after 250ns (T250), and the third from the final state after 1 microsecond of simulation (T1000). MD was executed in OpenMM 7.1.0 33 using a Langevin integrator with a 2fs timestep and a friction coefficient of 10/picosecond. BLUES simulations were run for 10,000 iterations with 2ps increments of MD between each Val111 dihedral state evaluation, repeated as needed until the rotamer fell within the biased dihedral range described previously. Each NCMC move was executed for 1004 steps with nprop = 3, such that NCMC steps were weighted toward the middle (lambda 0.2 to 0.8) of the NCMC switching protocol as described previously.