The extent to which water mass mixing and ocean ventilation contribute to nitrous oxide (N2O) distribution at the scale of oceanic basins is poorly constrained. We used novel N2O and chlorofluorocarbon measurements along with multiparameter water mass analysis to evaluate the impact of water mass mixing and Atlantic Meridional Overturning Circulation (AMOC) on N2O distribution along the Observatoire de la variabilité interannuelle et décennale en Atlantique Nord (OVIDE) section, extending from Portugal to Greenland. The biological N2O production has a stronger impact on the observed N2O concentrations in the water masses traveling northward in the upper limb of the AMOC than those in recently ventilated cold water masses in the lower limb, where N2O concentrations reflect the colder temperatures. The high N2O tongue, with concentrations as high as 16 nmol kg−1, propagates above the isopycnal surface delimiting the upper and lower AMOC limbs, which extends from the eastern North Atlantic Basin to the Iceland Basin and coincides with the maximum N2O production rates. Water mixing and basin‐scale remineralization account for 72% of variation in the observed distribution of N2O. The mixing‐corrected stoichiometric ratio N2O:O2 for the North Atlantic Basin of 0.06 nmol/μmol is in agreement with ratios of N2O:O2 for local N2O anomalies, suggesting than up to 28% of N2O production occurs in the temperate and subpolar Atlantic, an overlooked region for N2O cycling. Overall, our results highlight the importance of taking into account mixing, O2 undersaturation when water masses are formed and the increasing atmospheric N2O concentrations when parameterizing N2O:O2 and biological N2O production in the global oceans.