Grays Athletic have been refused entry to the Ryman Premier as the league have been unable to approve their groundshare with East Thurrock.
The Essex club had opted to drop down a level after being relegated from the Blue Square Premier last season.
A Ryman League statement read: "The conclusion has been reached that Grays does not meet criteria for membership."
The club had arranged to play at East Thurrock after the lease on their New Rec home was not extended.
Grays successfully appealed for a Ryman League spot in June after the Football Association said they had failed to submit the relevant paperwork on time.
Athletic now plan to appeal the latest decision but will face playing Essex Senior League football if unsuccessful.
However, like other Ryman Premier Division clubs, Grays have been given a bye until the first qualifying round of the FA Cup, and have also been included in the draw for the FA Trophy.
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