Former US President Donald Trump boards Air Force One before departing Harlingen, Texas. AFP
"The account referenced has been permanently suspended for violating our ban evasion policy," a Twitter spokesperson told AFP.
The account, @KhameneiSite, this week posted an animated video showing an unmanned aircraft targeting Trump, who ordered a drone strike in Baghdad two years ago that killed top Iranian commander General Qassem Soleimani.
Supreme leader Ayatollah Ali Khamenei's main accounts in various languages remain active. Last year, another similar account was suspended by Twitter over a post also appearing to reference revenge against Trump.
The recent video, titled "Revenge is Definite", was also posted on Khamenei's official website.
According to Twitter, the company's top priority is keeping people safe and protecting the health of the conversation on the platform.
The social media giant says it has clear policies around abusive behavior and will take action when violations are identified.
As head of the Quds Force, the foreign operations arm of Iran's Revolutionary Guards, Soleimani was the architect of its strategy in the Middle East.
He and his Iraqi lieutenant were killed by a US drone strike outside Baghdad airport on January 3, 2020.
Khamenei has repeatedly promised to avenge his death.
On January 3, the second anniversary of the strike, the supreme leader and ultraconservative President Ebrahim Raisi once again threatened the US with revenge.
Trump's supporters regularly denounce the banning of the Republican billionaire from Twitter, underscoring that accounts of several leaders considered authoritarian by the United States are allowed to post on the platform.