The 301 Queen Blue Night streetcar route operates between Neville Park Loop and Long Branch Loop, generally in an east-west direction.

One single service is operated:  the 301 (Neville Park-Long Branch) branch operates during the overnight period, seven days a week.

Westbound “301 LONG BRANCH” streetcars operate from Neville Park Loop (on the southwest corner of Queen Street East and Nursewood Road) via west on Queen Street East, west on Queen Street West, west on The Queensway, through Humber Loop, continuing southwest and west on Lake Shore Boulevard West to Long Branch Loop (on the north side of Lake Shore Boulevard West, west of Browns Line).

Eastbound “301 NEVILLE PARK” streetcars operate from Long Branch Loop via east and northeast on Lake Shore Boulevard West, through Humber Loop, continuing east on The Queensway, east on Queen Street West, and east on Queen Street East to Neville Park Loop.

All streetcars on the route serve local streetcar stops along the route.