The atmospheric boundary layer (ABL) flow in urban morphologies, ranging from micro-scale to local-scale, will become non-isothermal due to the combined effects of solar radiation, wall heat transfer, the urban heat island, and anthropogenic heat, resulting in a mutually coupled urban thermal and wind environment. An appropriate temperature inflow profile is the key prerequisite for Computational Fluid Dynamics (CFD) simulations of the non-isothermal ABL flow and the urban thermal and wind environment, as it must coincide with actual thermal characteristics while being compatible with CFD. Based on the equilibrium ABL model, a new equilibrium temperature inflow profile compatible with the energy equation, the standard gradient diffusion hypothesis, and the turbulence model is derived, which is proportional to the turbulent kinetic energy inflow profile, with the ratio further determined by the wall heat flux, demonstrating that turbulent thermal diffusion is strongly related to turbulent characteristics and atmospheric stability. To address the scaled effect, its full expression is provided based on thermodynamic similarity criteria. Using a numerical model of empty domain based on the SST k-ω model, the horizontal homogeneity of the equilibrium thermal and wind field modeled by the new set of inflow profiles is verified. Taking the flow around a single square building model as an example, its applicability in simulating the urban thermal and wind environment around buildings has been numerically validated, where the comparison of numerical and test results reveals tolerable errors overall.