The eviction process on Monday came after a group of American anti-war activists began moving into the embassy last month.
"The US does not recognise the authority of the former (Nicolas) Maduro regime or any of its former representatives, to allow any individuals to lawfully enter, remain on this property, or take any other action with respect to this property," said the evacuation order.
Guaido's envoys in Washington "have requested and directed anyone who is present on this property to depart from it immediately, and to not return without these ambassadors' express authorisation".
"Any person who refuses to comply with these requests and orders to depart from this property will be trespassing in violation of federal and District of Columbia law and may be arrested or criminally prosecuted," it added.
Medea Benjamin, one of the founders of the anti-war group Code Pink, had previously told CNN that diplomats from the government of Venezuelan President Nicolas Maduro had given them access to the embassy before they left as the standoff unfolded between Maduro and Guaido.
"The government of the Bolivarian Republic of Venezuela has not authorised the entry of police officers into the former Embassy building in Washington, DC. This intrusion is yet another violation of international law by US authorities and an aggression against Venezuela," Ron, a Maduro official, said in a tweet.
Although some activists have already left the embassy, at least four remained inside, according to Efe news.
There were also pro-Maduro activists outside the building who support those who remain inside the compound.
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