Here, the electrostatic interactions are annihilated in both the solution and vacuum environments. For the vdW interactions, we decouple the molecule's interaction only in solution, thus removing the need to perform the calculation in vacuum. For both electrostatics and vdW interactions, we allow Yank to automatically determine the number of λ points with the trailblaze algorithm from Rizzi et al. (unpublished). We run Yank with Hamiltonian replica exchange over 2000 iterations with a time step of 2 fs. The analysis was done using Yank's analysis framework, which employs the multistate Bennett acceptance ratio (MBAR) method to recover the free energy from the MD trajectories.