In urban karst areas, such as the City of Bowling Green, Kentucky and the Tampa Bay Metropolitan Area, groundwater quality faces a variety of threats. The development of residential, commercial, and industrial landuse types allows for a wide variety of groundwater pollutants to enter the karst groundwater systems. Various different models and indices have attempted evaluative approaches to identify issues in urban karst areas, but the methods vary by location and lack a focus on urban karst groundwater quality. There also exists a lack of a data-driven approach that is able to capture short- and long-term changes in threats to groundwater quality as a result of urbanization. The overall purpose of this study was to develop a holistic, data-driven evaluation toolbox with threat, vulnerability, and monitoring assessment tools for urban karst groundwater systems to better determine the possible threats, data collection needs, monitoring parameters, and analytical approaches needed to ensure groundwater quality is maintained in urban karst regions. This study focused on: 1) determining what indicators, parameters, resolution, and data quality need to be prioritized to create an effective, holistic monitoring framework for urban karst groundwater, and 2) developing an effective assessment and evaluative tools for urban karst groundwater quality sites using historic and modern data in an urban karst setting. The outcomes include an Urban Karst Aquifer Resource Evaluation (UKARE) Toolbox with a Threat, Vulnerability, and Monitoring evaluation tools that were applied and validated through application of the Toolbox using case studies in the City of Bowling Green, Kentucky and the Tampa Bay Metropolitan Area in Florida. The results demonstrate the universal applicability of the UKARE Toolbox to different urban karst sites and its effectiveness at scoring for threats and vulnerabilities, as well as identifying potential monitoring sites through primary data collection of water quality parameters and emerging pathogens at over 150 sites between both study areas. The final results of this study are useful to develop monitoring and management plans through a standardized scoring and evaluation tool in order to influence urban karst groundwater monitoring and management.