To date, few isotope age constraints on primary oil migration have been reported. Here we present U-Pb dating and characterization of two fracture-filling, oil inclusion-bearing calcite veins hosted in the Paleocene siliciclastic mudstone source rocks in Subei Basin, China. Deposition age of the mudstone formation was estimated to be ca. 60.2–58.0 Ma. The first vein consists of two major phases: a microcrystalline-granular (MG) calcite phase, and a blocky calcite phase, each showing distinctive petrographic features, rare earth element patterns, and carbon and oxygen isotope compositions. The early MG phase resulted from local mobilization of host carbonates, likely associated with disequilibrium compaction over-pressuring or tectonic extension, whereas the late-filling blocky calcite phase was derived from overpressured oil-bearing fluids with enhanced fluid-rock interactions. Vein texture and fluorescence characteristics reveal at least two oil expulsion events, the former represented by multiple bitumen veinlets postdating the MG calcite generation, and the latter marked by blue-fluorescing primary oil inclusions synchronous with the blocky calcite cementation. The MG calcite yields a laser ablation–inductively coupled plasma–mass spectrometry U-Pb age of 55.6 ± 1.4 Ma, constraining the earliest timing of the early oil migration event. The blocky calcite gives a younger U-Pb age of 47.8 ± 2.3 Ma, analytically indistinguishable from the U-Pb age of 46.5 ± 1.7 Ma yielded by the second calcite vein. These two ages define the time of the late oil migration event, agreeing well with the age estimate of 49.7–45.2 Ma inferred from fluid-inclusion homogenization temperature and published burial models. Thermodynamic modeling shows that the oil inclusions were trapped at ~27.0–40.9 MPa, exceeding corresponding hydrostatic pressures (23.1–26.7 MPa), confirming mild-moderate overpressure created by oil generation-expulsion. This integrated study combining carbonate U-Pb dating and fluid-inclusion characterization provides a new approach for reconstructing pressure-temperature-composition-time points in petroleum systems.