Adding nitrogen fertilizers to agricultural soils contributes to increasing concentrations of nitrous oxide (N2O) in the atmosphere. However, the impacts of N addition on soil organic matter (SOM) turnover, SOM availability, and the ensuing SOM‐derived N2O emissions remain elusive. Within this context, the net change in direction and rate of SOM‐derived N2O production triggered by added N is termed the N2O priming effect. This incubation study examined the sources and priming of N2O production as a function of urea addition and multiple moisture contents in a soil with high SOM (55 g organic C kg−1). We assessed four water‐filled pore space (WFPS) conditions: 28, 40, 52, and 64%. Relative to controls receiving no N, urea addition increased N2O production by 2.6 times (P < .001). Cumulative N2O production correlated well with nitrification rates (r = .75; P = .03). We used 15N‐labeled urea to trace the added urea into N2O. Of the N added via urea, the recovery as N2O–N shifted from 0.02 to 0.17% when WFPS increased from 28 to 64% (P < .05). We also partitioned the N2O production into urea vs. SOM sources. More N2O was sourced from SOM than urea, with 59 ± 2% N2O originating from SOM. The magnitude of SOM‐derived N2O under urea was larger than that of the control, revealing that positive N2O priming was triggered by urea addition. Upon subtracting the controls, the primed N2O was a consistent 19 ± 2% of the total N2O produced by urea‐amended soils. Nevertheless, the priming magnitude rose sharply with increasing moisture by more than one order of magnitude from 4 to 48 μg N2O–N kg−1 soil and in exponential mode (R2 = .98). Soil moisture, SOM, and nitrification interacted to drive the sources and priming of N2O.