Migrating cancer cells release diverse extracellular vesicles (EVs) that carry bioactive cargo capable of influencing tumor progression and intercellular communication. However, the mechanisms by which genomic DNA and other intracellular components are incorporated into EVs during migration remain poorly understood. Here, we identify and characterize a previously undescribed population of large EVs (2–5 μm), termed Motile Vesicles (MoVes), that are generated during active cancer cell migration and associated with epithelial–mesenchymal transition (EMT). Live-cell imaging and scanning electron microscopy reveal that MoVes arise from trailing-edge retraction during mesenchymal migration and are actively shed by cancer cells but not by non-transformed cells. Whole-genome sequencing demonstrates that MoVes contain genomic DNA spanning the genome of the parent cell, recapitulating its mutational landscape. Comparative analyses across EV populations show that MoVes display distinct ultrastructural features, molecular composition, and sedimentation behavior. Proteomic profiling identifies enrichment of proteins associated with EMT and organelle biology, including the EMT regulator FAM3C, and ultrastructural analyses reveal incorporation of intracellular organelles such as mitochondria. Functional assays further demonstrate that MoVes-associated mitochondrial cargo can enhance oxidative phosphorylation in recipient cells. Together, these findings identify MoVes as a migration-associated EV population that carries genomic DNA and organelle cargo during cancer cell migration, revealing a previously unrecognized mechanism of extracellular DNA and organelle dissemination in cancer.