Identifying the tectonic transition from oceanic subduction to collision is crucial for tracking the final stage evolution of ancient orogenic belts. In this study, we present new geochronological and geochemical data for the Mozbaysay mafic–ultramafic complex in the Balikun area, eastern North Tianshan of the southern Central Asian Orogenic Belt. This complex had intruded into the late Carboniferous volcano-sedimentary rocks and is comprised mainly of hornblende gabbro and lherzolite. Zircon U-Pb ages of the hornblende-gabbros reveal that this complex was emplaced at ca. 305 Ma. Geochemical analyses suggest these mafic–ultramafic rocks are characterized by slight enrichment of light rare earth elements (LREEs) and relatively depleted heavy rare earth elements (HREEs), resembling enriched mid-ocean ridge basalt (E-MORB). They also exhibit restricted (87Sr/86Sr)i ratios (0.702396–0.704295) and εNd(t) values (+7.0 to +9.1), indicative of a depleted mantle source with minimal crustal contamination. Incompatible element ratios (i.e., Nb/Ta, Zr/Hf, Rb/Nb, and Ba/Nb) suggest the involvement of subducted slab-derived aqueous fluids in their mantle source. These collectively indicate that the parental magmas of the Mozbaysay mafic–ultramafic rocks may have been generated by a mixed mantle source consisting of the E-MORB-like asthenospheric mantle, subcontinental lithospheric mantle (SCLM), and hydrous fluids from subducted slab. Furthermore, a slab break-off model is proposed to explain the generation of these latest Carboniferous mafic–ultramafic rocks. Integrating these findings with regional geological data, we propose that the tectonic transition from subduction (slab roll-back) to collision (slab break-off) along the Kalamaili suture zone occurred at ca. 305–300 Ma.