Karst hydrologic systems are important resources in the state of Tennessee both as drinking water resources and as centers for possible biological diversity. These systems are susceptible to contamination due to the inherent connectivity between surface water and groundwater in karst landscapes. A partnership between the U.S. Geological Survey (USGS) and Tennessee Department of Conservation (TDEC) was formed to investigate karst spring systems across the state utilizing fluorescent groundwater tracing, particularly in areas where these resources may be used as drinking water sources. In fall 2021, USGS and TDEC staff identified possible vulnerabilities or complexities that may exist within karst spring systems based upon maturity of karst development, underlying geology, and uncertainties related to estimated recharge areas. Based upon initial research, several study areas were selected. In late winter 2021, fieldwork began in areas surrounding the Tennessee communities of Cowan, Jasper, Vanleer, and Woodbury. These communities are in three physiographic provinces; Cowan and Jasper are near the Cumberland Plateau while Vanleer and Woodbury are located on the Western and Eastern Highland Rim, respectively. These systems are in areas where the hydrology has been significantly altered by karst processes and thus the groundwater pathways are complex and unpredictable. Data within each child item of this data release are named with a two-letter abbreviation unique for the community where the tracing occurred and the water year the work was conducted (e.g. WD23). Abbreviations for the communities are as followed: CW = Cowan, JS = Jasper, VN = Vanleer, WD = Woodbury. This data release contains shapefiles that relate to dye injection locations, monitoring sites, and dye traces. All files were created in ArcGIS Pro and each shapefile contains associated attributes for the features contained within. Layer files are included with the datasets to match symbology found in figures in the accompanying report. All shapefiles and layers were created and modified in ArcGIS software. For a full description of the methods to create these files, see Process Steps in "AllData22_metadata.xml."