It runs 163.1 kilometres (101.3 mi) from Vienna, the Austrian capital, to the border with the Czech Republic at Gmünd.

The southern terminus of the line is at Wien Franz-Josefs-Bahnhof, in the Alsergrund district of Vienna.

The S40 operates at hourly intervals between Wien Franz-Josefs-Bahnhof and St. Polten, leaving the Franz-Josefs-Bahn at Tulln.

[3] Direct service between Vienna and Prague over the route, defunct since 1992, resumed in December 2022 as the Silva Nortica (lit.

[4] Regional-Express trains operate hourly to Krems an der Donau, crossing the Danube at Tulln and leaving the Franz-Josefs-Bahn at Absdorf-Hipperdorf.