The cosmopolitan inversion In(3R)Payne in Drosophila melanogaster decreases in frequency with increasing distance from the equator on three continents, indicating it is subject to strong natural selection. We investigated patterns of genetic variation and linkage disequilibrium (LD) in 24 molecular markers located within and near In(3R)Payne to determine if different parts of the inversion responded to selection the same way. We found reduced variation in the markers we used compared to others distributed throughout the genome, consistent with the inversion having a relatively recent origin (<N(e) generations). LD between markers and In(3R)Payne varied significantly among markers within the inversion, with regions of high association interspersed by regions of low association. Several factors indicate that these patterns were not due to demographic factors such as admixture and bottlenecks associated with colonization, but instead reflected strong epistatic selection. Furthermore, we found that nonadjacent regions with high association to the inversion contained markers with the strongest clinal patterns in allele frequency; in most cases, the level of clinal variation was beyond what could be explained by hitchhiking with In(3R)Payne, indicating that genes within these regions are targets of selection. Our results provide some support for the hypothesis that inversions persist in natural populations because they hold together favorable combinations of alleles that act together to facilitate adaptive shifts.