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The Florida Wildlife Corridor is increasingly affected by intensifying human pressures, leading to cascading environmental changes that threaten ecological integrity and connectivity. To inform conservation strategies and land management decisions, we developed a geographically explicit statewide Human Modification Index (HMI) for Florida. This index captures the extent of anthropogenic pressure across the landscape, incorporating infrastructure development, population density, transportation networks, energy production, agriculture, recreational use, and pollution. Using three distance-based kernel categories, we modeled the spatial influence of human activities on surrounding landscapes. We integrated the HMI with a Landscape Integrity layer—representing ecological condition based on land cover and road fragmentation—and applied the Omniscape algorithm to model cumulative and normalized current flow, highlighting both broad connectivity patterns and constrained, channelized movement areas known as “pinch points”. We then developed a novel composite index by multiplying these layers with potential habitat richness and natural community data to identify areas of high ecological value and landscape connectivity. The top 20% of composite scores defined high-priority areas, which were further evaluated using tax parcel data to estimate just value and land value. We focused on ten critical linkages within Priority 1 areas of the Florida Wildlife Corridor to (1) map connectivity across ten critical linkages, (2) identify “pinch-point” bottlenecks with high biodiversity and natural community scores, and (3) quantify the economic value of key areas. High-value composite pinch points occupy just 70,400 ha (1.6% of 4.45 M ha of linkages) yet comprise 57% of top tier cumulative connectivity flow in the critical linkages. The results demonstrate substantial variability in connectivity, vulnerability, and economic stakes across the corridor, emphasizing the strategic importance of conserving pinch points to maintain landscape permeability. Our study presents a novel, data-driven framework that integrates ecological integrity and land valuation to inform actionable insights for protecting Florida’s most critical wildlife corridors.