The referendum on whether or not the UK should remain in the European Union will not take place on the same day as national and local elections next May.
Downing Street said ruling out 5 May was a concession to MPs’ concerns.
Opponents said it would have “confused the issue” to have the vote on the same day as other elections.
The government is also expected to address concerns over the “purdah” period, which restricts campaigning before a referendum is held.
Speaking after the amendment was tabled on Monday, the Number 10 spokesman said: “We have listened to the views expressed by MPs across the house and decided that we won’t hold the referendum on 5 May 2016.”
That date is the same day elections to the Scottish Parliament and Welsh and Northern Irish assemblies are being held, along with the London mayoral election.
Elections will also be held that day in 126 English local authorities, and all Welsh and Scottish councils.
Conservative MP Bernard Jenkin welcomed the ruling out of the 5 May date, saying it would have “confused the whole issue” holding the poll on the same day as other major elections.
“It would have obscured the clear choice,” he told BBC Radio 4’s Today programme.
By BBC Newsnight’s Allegra Stratton and James Clayton
There had been a move among the prime minister’s team to bring the poll forward.
Downing Street advisers argued that staging the referendum poll on the same day as next year’s local and mayoral elections could drive up turnout for the referendum among some of Britain’s most pro-European regions.
They also wanted the prime minister to stage an early referendum to allow the Conservative party to capitalise on goodwill, after winning its first majority in 23 years.
Eurosceptic MPs were dismayed at the idea, believing the prime minister was trying to rush the process.
Opposition politicians, including the SNP’s foreign affairs spokesman Alex Salmond, argued that such an important decision should not be held on the same day as other elections.
Downing Street has also said the government will “seek to address” the concerns of Tory Eurosceptics who have tabled amendments to the EU Referendum Bill to reinstate the “purdah” period.
The Political Parties, Elections and Referendums Act 2000 set out a 28-day period ahead of a referendum, during which ministers, government departments and local authorities are banned from publishing material relating to the issue in question.
But Prime Minister David Cameron has said this would mean ministers were barred from speaking about European court decisions and the EU budget.
He has also said the government will not be “neutral” when it comes to the in/out referendum.
Eurosceptic former cabinet minister Owen Paterson said it would be “unacceptable” for the government to use public money to promote a vote to stay in the EU.
Mr Jenkin said the existing purdah rules had been applied to every referendum staged in the past 15 years and had been endorsed by both the Electoral Commission and the Committee on Standards in Public Life.
“I think the government has been very badly advised on the date and on this. If they had accepted the Electoral Commission’s advice on purdah we would not be in this mess. The Electoral Commission thinks the purdah period should be longer than 28 days.
“The government thinks they should be able to use their civil servants and their press officers and their special advisers to support their activities…These are rules that the government is scrapping and people will think they are trying to rig the whole playing field.”
Mr Jenkin said the Conservative rebels would not withdraw their amendment and it was up to the government to table its own amendments later in the bill’s passage if they want to address other concerns.
But he dismissed suggestions a backbench Tory rebellion over the issue was the equivalent of a “Maastricht moment”, a reference to the bitter battles between Tory backbenchers and John Major’s government over Europe which threatened its very survival in mid-1990s.
“This is not even about Europe,” he added. “It is about whether you want a fair referendum. We are all very happy with the prime minister’s policy of ‘negotiate and decide’ on the basis of a referendum.”
Downing Street said Mr Cameron wanted the ability to publish material but was not seeking to “overly influence” the outcome of the campaign.
Last week, MPs overwhelmingly backed plans for a referendum, allowing the legislation to move to the next stage of its progress through Parliament. It still has several more stages to pass through, however.
It enters its Committee stage on Tuesday, during which it will be debated in detail by the whole of the House of Commons.