Human commensal species such as rodent pests are often widely distributed across cities and threaten both infrastructure and public health. Spatially-explicit population genomic methods provide insights into movements for cryptic pests that drive evolutionary connectivity across multiple spatial scales. We examined spatial patterns of neutral genome-wide variation in brown rats (Rattus norvegicus) across Manhattan, New York City (NYC) using 262 samples and 61,401 SNPs to understand: 1) relatedness among nearby individuals and the extent of spatial genetic structure in a discrete urban landscape; 2) the geographic origin of NYC rats, using a large, previously-published dataset of global rat genotypes; and 3) heterogeneity in gene flow across the city, particularly deviations from isolation-by-distance. We found that rats separated by ≤200m exhibit strong spatial autocorrelation (r = 0.3, p = 0.001) and the effects of localized genetic drift extend to a range of 1400m. Across Manhattan, rats exhibited a homogeneous population origin from rats that likely invaded from Great Britain. While traditional approaches identified a single evolutionary cluster with clinal structure across Manhattan, recently-developed methods (e.g., fineSTRUCTURE, sPCA, EEMS) provided evidence of reduced dispersal across the island's less residential Midtown region resulting in fine-scale genetic structuring (FST = 0.01) and two evolutionary clusters (Uptown and Downtown Manhattan). Thus, while some urban populations of human commensals may appear to be continuously distributed, landscape heterogeneity within cities can drive differences in habitat quality and dispersal, with implications for the spatial distribution of genomic variation, population management, and the study of widely distributed pests. This article is protected by copyright. All rights reserved.