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The mosaic depicts the the ancient Greek music master Orpheus and lyre resting on his left knee, his hunting dog alongside him and a myriad of beasts all around him.
The animals include a tiger, leopard, lion, elephant, bear, gryphon, stag, horse, wild boar and birds such as pheasants, peacocks and doves.
A pair of water nymphs are depicted in each spandrel.
The work is currently on display at Prinknash Abbey, near Stroud, but the lease has come to an end and its owner - who does not wish to be identified - has decided to sell.
Iona Sale, who is publicising the event for auctioneers Chorley's, said it had been challenging putting an estimated sale price on the mosaic - because no-one had ever attempted such a feat before.
Bob Woodward turned from builder into research scholar as he tried to discover what the missing parts must have looked like.
The earliest report he found dated back to 1693, when Celtic scholar Edward Llwyd recorded having seen ‘birds and beasts on the floor’.
He conducted careful research into other Orpheus mosaics to help replace the missing sections as accurately as possible, particularly the Barton Farm mosaic at Cirencester which had several similar beasts.