The B-1B Lancer passed over the Gulf, Bab al-Mandeb Strait, Suez Canal and Gulf of Oman, the US Central Command, or Centcom, said in a statement Saturday.
It also flew over the Strait of Hormuz, a chokepoint for a fifth of world oil output at the head of the Gulf that Iran considers a strategic area of influence.
"The bomber task force mission... was intended to deliver a clear message of reassurance," Centcom said.
Fighter jets from Israel, Saudi Arabia and Bahrain, all US allies opposed to the Islamic Republic of Iran, escorted the US bomber over their respective airspaces.
"Military readiness for any contingency or mission –- from crisis response to multilateral exercises to one-day presence patrols like this –- depend on reliable partnerships," said Centcom commander General Frank McKenzie, the head of US forces in the Middle East.
Egyptian fighter jets also accompanied the B-1B, a supersonic bomber that can carry the heaviest conventional payload of all the US military's planes.
In January, a US B-52 bomber, also capable of carrying nuclear weapons, flew over the Middle East.
Since February, Iran and Israel have been accused of engaging in what analysts have called a "shadow war", in which vessels linked to each nation have come under attack in waters around the Gulf in tit-for-tat exchanges.
The regional patrol, Centcom's fifth such operation this year, comes as talks aimed at reviving the 2015 deal to limit Iran's nuclear program have stalled.
US President Joe Biden has repeatedly offered to return to the nuclear accord, but his administration has voiced growing frustration over delays after a hardline government took office in Iran.
Then-president Donald Trump withdrew the United States from the deal in 2018 and imposed sweeping sanctions, leading Iran to step up contested nuclear work in retaliation.
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