[6] The popular tourist route mostly follows the southern bank of the Murray River and effectively acts as the northernmost highway in Victoria.

Murray Valley Highway commences at the intersection with Sturt Highway just outside Euston, New South Wales and heads south to cross the Murray River over the Robinvale-Euston bridge at Robinvale and into Victoria; the western end of route B400 starts here.

The highway continues in a south-easterly direction, tracking close to the southern bank of the Murray River for the majority of its length through the towns of Swan Hill, Kerang, Cohuna, Echuca, Nathalia, Strathmerton, Cobram, Yarrawonga and Rutherglen, until it reaches Wodonga, before heading in an easterly direction via Tallangatta until it eventually terminates at the foothills of the Great Dividing Range at Corryong; the road beyond crosses the border east into New South Wales as Alpine Way, to eventually reach Khancoban and Jindabyne.

It becomes hillier and more winding east of Wodonga, with a moderately steep mountain pass near Shelley, midway between Tallangatta and Corryong.

[16][17] The alignment was further altered at both ends in 1990: The passing of the Road Management Act 2004[20] granted the responsibility of overall management and development of Victoria's major arterial roads to VicRoads: in 2004, VicRoads re-declared the road as Murray Valley Highway (Arterial #6570), beginning at the New South Wales border at Robinvale and ending at the New South Wales border in Towong Upper.