Habitat loss and fragmentation are the major causes of biodiversity loss, and, increasingly, habitat is fragmented by urbanization. Yet, the degree to which urbanization creates barriers to animal dispersal remains poorly understood. We used population genetic techniques to determine whether urbanization and/or natural landscape features are dispersal barriers to a butterfly, Atrytonopsis new species 1, throughout its range on coastal sand dunes that are increasingly threatened by development. Using AFLP markers that produced 89 polymorphic loci, we found significant population structure across the range of Atrytonopsis sp1. We found no indication that existing levels of urbanization were barriers to Atrytonopsis sp1 dispersal. Rather, two natural barriers, an ocean inlet and maritime forest, explained the genetic structure. Even in areas with long histories of urbanization, we found no significant isolation-by-distance relationship, and there was very low genetic differentiation between sampling locations. Consequently, conservation strategies for Atrytonopsis sp1, and potentially for other mobile insects that use open-structured habitats, should not focus explicitly on habitat corridors through urban areas, but rather should seek to preserve and restore as much habitat as possible across the butterfly's range.