Public water supply systems (PWSS) are critical infrastructure that are vulnerable to contamination and physical disruption. Exploring susceptibility of PWSS to such perturbations requires detailed knowledge of supply system structure and operation. The physical structure of the distribution system (i.e., pipeline connections) and basic information on sources are documented for most industrialized metropolises. Yet, most information on PWSS function comes from hydrodynamic models that are seldom validated using observational data. In developing regions, the issue may be exasperated as information regarding the physical structure of the PWSS may be incorrect, incomplete, undocumented, or difficult to obtain in many cities. Here, we present a novel application of stable isotopes in water (SIW) to quantify the contribution of different water sources, identify static and dynamic regions (e.g., regions supplied chiefly by one source vs. those experiencing active mixing between multiple sources), and reconstruct basic flow patterns in a large, complex PWSS. Our analysis, based on a Bayesian mixing model framework, uses basic information on the SIW and production volumes of sources but requires no information on pipeline connections in the system. Our work highlights the ability of SIW to analyze PWSS and document aspects of supply system structure and operation that can otherwise be challenging to observe. This method could allow water managers to document spatiotemporal variation in flow patterns within PWSS, validate hydrodynamic model results, track pathways of contaminant propagation, optimize water supply operation, and help monitor and enforce water rights.