This dataset comprises integrated quantitative and qualitative experimental data generated for the characterization of a putative anti-eukaryotic effector associated with the Type VI Secretion System (T6SS) of Xanthomonas citri subsp. citri. The data were obtained using a combination of molecular biology, biochemical, NGS, and imaging-based approaches, including blotting techniques, and microscopy. Quantitative data include measurements such as area, time, signal intensity, and relative abundance of biological molecules, while qualitative data consist of microscopy images and experimental videos. Experiments were conducted using the bacterial model X. citri subsp. citri and the eukaryotic model organisms Dictyostelium discoideum and Saccharomyces cerevisiae, enabling the assessment of effector activity across different biological systems. Whenever applicable, experiments were performed in biological triplicates to ensure statistical robustness. Data acquisition was carried out using multi-user equipment from the Institute of Biology at the University of Campinas (IB-UNICAMP), with additional use of facilities like National Institute of Photonics Applied to Cell Biology (INFABiC), Laboratório Central de Tecnologias de Alto Desempenho em Ciências da Vida (LaCTAD), and NGS soluções genômicas. Metadata associated with the dataset include title, subject, author information, abstract, technical specifications of the equipment used, keywords, and persistent identifiers, following the DataCite Metadata Schema standard. The dataset was stored in multiple file formats according to data type, including spreadsheets (XLS/XLSX), tabulated text files (TXT, CSV, TSV), image and video formats (JPEG, TIFF, PNG, MP4, WMV, OGV), and statistical analysis files (RData). The data cannot be made available for intellectual property reasons. For more information, please contact those responsible.