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It was opened in and was partly owned by actor Johnny Depp ; the other part owner was Sal Jenco who starred in 21 Jump Street with Depp. In early , Australian singer Jason Donovan suffered a drug-induced seizure at the club and survived; the Viper Room has undergone several changes in ownership, and continues to host music of multiple genres, including metal , punk rock , and alternative rock. While predominantly known as a music venue, the Viper Room also hosts a lower level below the stage and audience area, which is home to a large whiskey bar.
The location was originally a grocery store, from into the s. In the s it was converted into a nightclub called the "Cotton Club", entirely unrelated to the Harlem original; this was soon replaced by "The Greenwich Village Inn", the "Rue Angel" and finally "The Last Call" during the s. From , the location was a bar called "The Melody Lounge". In or , it became "Filthy McNasty's". Weiss , who had performed there for years, suggested to Depp that they revitalize the spot and rename it "The Viper Room".
At Depp's request, Tom Petty and the Heartbreakers performed on the club's opening night;  the Viper Room continues to frequently host metal and punk rock bands. In the film Valley Girl , the building then housing a nightclub called The Central was used for scenes featuring the new-wave band the Plimsouls. London Fog was a lesser-known nightclub half way up the same block from the Whisky a Go Go where the Doors had their first regular gigs for four months in early Several scenes in the John Travolta film Be Cool were set at or in the club,   although only the exterior of the site was used in the shoot.
The Viper Room is also featured in the documentary DiG! As part of the settlement of a lawsuit involving the disappearance of co-owner Anthony Fox in , Depp relinquished his ownership of the Viper Room in ;  the club changed hands multiple times between and ; the club is currently owned by Viper Room Holdings, Ltd.
The location has been featured on the season 14 episode of paranormal series Ghost Adventures as one of their lockdown locations, where they investigated the building for two days; the episode categorizes in detail the death of actor River Phoenix believing they came into contact with his spirit and shows video evidence of EVP. They also captured a guitar playing by itself on the stage, believed to be the spirit of former owner Anthony Vivien Fox, a singer that has been missing and last seen on December 19, The Viper Room is the setting for the initial set of poker games Molly Bloom hosted.