By virtue of a biomimetic nanopore and single-walled carbon nanotubes, a new sensor for adenosine triphosphate (ATP) detection is designed. As compared to the routine approach, the present scenario does not entail the surface modification of nanopore with analyte-specific probes. The underlying mechanism relies on a symmetric nanopore sequentially modified with polyethyleneimine (PEI) and Zr(4+) that can quantitate the concentration of ATP-bound aptamer, while other free aptamers are removed by single-walled carbon nanotubes (SWNTs). The detection limit of the nanopore sensor is 27.46 nM, and the linear range is from 50 nM to 400 nM. The biosensor with an excellent selectivity against guanosine triphosphate (GTP), uridine triphosphate (UTP), and cytosine triphosphate (CTP) can be applied in the real samples such as Hela cell.