National Route 1 (officially PY01, in Spanish, Ruta Nacional Número 1, or simply Ruta Uno) is one of the most important and one of the first highways in Paraguay, which runs from the capital city, Asunción to Itapua Department's capital, Encarnación. Crossing the departments of Central, Paraguarí, Misiones and Itapua. It starts on the intersection of Choferes del Chaco and Fernando de la Mora Avenues, known as Cuatro Mojones, and ends at the San Roque González de Santa Cruz Bridge, on the Paraná River, traversing 382 km (237 mi).

The following table shows the distances traversed by PY01 in each different department, and important cities that it passes by (or near).

Km	City	Department	Junctions/Tolls
0	Asunción	Capital District
8	Villa Elisa/Fernando de la Mora	Central
13	Ñemby/San Lorenzo	Central
19	San Antonio	Central
23	Ypané	Central
30	Guarambaré	Central
37	Itá	Central
48	Yaguarón	Paraguarí
69	Paraguarí	Paraguarí	PY10
83	Carapeguá	Paraguarí	PY18
95	San Roque G. de Santa Cruz	Paraguarí
109	Quiindy	Paraguarí
138	Caapucú	Paraguarí
160	Villa Florida	Misiones
178	San Miguel	Misiones
196	San Juan Bautista	Misiones
224	San Ignacio	Misiones	PY04
243	Santa Rosa	Misiones
252	San Patricio	Misiones	PY20
288	General Delgado	Itapúa
320	Coronel Bogado	Itapúa	PY08/
330	Carmen del Paraná	Itapúa
365	San Juan del Paraná	Itapúa
382	Encarnación	Itapúa	PY06