Divine Playhouse opened in the former Catholic Church at 420 Kent Street in Sydney on July 8, attracting protests on its opening night.
Leaseholders Heaps Gay Events say its lease agreement for the venue was then terminated last Friday with no attempt at mediation.
The queer club and arts hub began clearing out late on Wednesday after holding only three events.
"... the trade carried on by HG Events has insulted and mocked the sincerely held religious beliefs of millions of Christian Australians," reads a legal complaint from the property's landlords sent on July 9.
Images from events at the venue include a photo of a performer dressed as a pig, satirising the rite of holy communion by handing out French fries.
The NSW offensive trade laws under which it is being forced out date back to 1842 and are intended to regulate foul smell and waste rather than morality, according to Heaps Gay Events.
It has not confirmed if the site has been vacated and has not provided requested documents confirming it has been formally evicted.
Heaps Gay Events says the amount spent on the short-lived venue has exceeded $200,000 including legal costs, which reportedly includes NSW government and council funding.
Venue owners have been contacted for comment through HWLE Lawyers.
A petition in support of the leaseholders has attracted almost 19,000 signatures while an online crowdfunding campaign has raised $42,000.
Petition organisers claim opposition to the venue has been co-ordinated by far-right Christian groups and influencers, and the controversy raises questions about artistic freedom.
"If this can happen to Divine Playhouse, it creates a dangerous precedent for independent arts venues, artists and minority communities across Australia," the petition reads.
Another petition in support of the closure has just under 8000 signatures.
The site was deconsecrated in 1932 and had also been the longtime home of the Genesian Theatre Company.
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