Ticks and tick-borne pathogens represent the greatest vector-borne disease threat in the United States. Blacklegged ticks are responsible for most human cases, yet the disease burden is unevenly distributed across the northern and southern United States. Understanding the genetic characteristics influencing phenotypic differences in tick vectors is critical to elucidating disparities in tick-borne pathogen transmission dynamics. Applying evolutionary analyses to molecular variation in natural tick populations across ecological gradients will help identify signatures of local adaptation, which will improve control and mitigation strategies. In this study, we performed whole genome nanopore sequencing of individual (