The early colonial period witnessed new scales of connectivity and unprecedented projects of resource extraction across the Spanish Americas. Yet such transformations also drew heavily on preexisting Indigenous landscapes, technologies, and institutions. Drawing together recent discussions in archaeology and geography about mobility and resource materialities, this article takes the early colonial route as a central object of investigation and contributes to new emerging interpretive frameworks that make sense of Spanish colonialism in the Americas as a variable, large-scale, and materially constituted process. Using three case studies—the ruta de Colón on the island of Hispaniola, the routes connecting the southeastern Caribbean islands with mainland South America, and the ruta de la plata in the south-central Andes—we develop a comparative archaeological analysis that reveals divergent trajectories of persistence, appropriation, and erasure in the region's routes and regimes of extraction and mobility during the fifteenth and sixteenth centuries.