In a dark room in the earliest days of settlement in Melbourne, a gold nugget and a lice comb fall unnoticed through a crack in the floorboards.

For more than 150 years they were forgotten in the gloom, until unearthed from the rubble of the Mistletoe Hotel as part of a major archaeological dig in the CBD.

The relics were discovered in one of the most significant archaeological digs in Victoria's history. Around 250,000 artefacts have been excavated from the former pub site in MacKenzie Street and dated back to within 20 years of Melbourne's founding by European settlers.

The collection is the state's largest assemblage of business-related artefacts, preserved for the last 55 years under an asphalt car park. Previous large excavations, such as the Little Lon dig, have focused on former residential areas.