Significance The quest for an optimal xylose pathway in yeast is of utmost importance along the way to realizing the potential of lignocellulosic biomass conversion into fuels and chemicals. An often-overlooked aspect of this catabolic pathway is the molecular transporter of this sugar. Here we demonstrate that sugar transport preference and kinetics can be rewired through the programming of a specific sequence motif. The result is a study to rationally alter the sugar preference of a protein through defined sequence-level modifications. In these cases, primary hexose transporters were rewired into xylose transporters.