Water resource management requires knowledge of the natural variability in streamflow over multiple time scales. Reconstructions of streamflow derived from moisture-sensitive trees extend, in both time and magnitude, the variability provided by relatively short gage records. In this study, we present a network of 14 annual streamflow reconstructions, 300–600 years long, for gages in the Upper Colorado and South Platte River basins in Colorado generated from new and existing tree-ring chronologies. Gages for the reconstruction were selected on the basis of their importance to two of the largest Colorado Front Range water providers, who provided the natural flow data for the calibration with tree-ring data. The reconstruction models explain 63–76% of the variance in the gage records and capture low flows particularly well. Analyses of the reconstructions indicate that the 20th century gage record does not fully represent the range of streamflow characteristics seen in the prior two to five centuries. Multi-year drought events more severe than the 1950s drought have occurred, notably in the 19th century, and the distribution of extreme low flow years is markedly uneven over the past three centuries. When the 14 reconstructions are grouped into Upper Colorado, northern South Platte, and southern South Platte regional flow reconstructions, the three time series show a high degree of coherence, but also time-varying divergences that may reflect the differential influence of climatic features operating in the western U.S. These reconstructions are currently being used by water managers to assess the reliability of water supply systems under a broader range of conditions than indicated by the gage records alone.