Let R be a commutative Noetherian ring. We prove that if R is either an equidimensional finitely generated algebra over a perfect field, or an equidimensional equicharacteristic complete local ring with a perfect residue field, then the annihilator of the singularity category of R coincides with the Jacobian ideal of R up to radical. We establish a relationship between the annihilator of the singularity category of R and the cohomological annihilator of R under some mild assumptions. Finally, we give an upper bound for the dimension of the singularity category of an equicharacteristic excellent local ring with isolated singularity. This extends a result of Dao and Takahashi to non-Cohen–Macaulay rings.