DENVER — If Democrats assumed Vermont Sen. Bernard Sanders would fall in line after losing the presidential nomination, they were sadly mistaken.

Mr. Sanders unveiled late Wednesday the launch of Our Revolution, a campaign organization aimed at promoting progressive issues, by picking a fight with the Colorado Democratic establishment over the state’s hotly contested single-payer health care ballot initiative.

He endorsed Amendment 69, known as ColoradoCare, even though top Democrats have mobilized against the sweeping $25 billion universal health care proposal amid fears that it will drag down Democratic candidates in November.

“There is an important ballot initiative in Colorado which calls for a [Medicare-for-All] health care system,” Mr. Sanders said in a speech from Burlington, Vermont, which was seen by thousands gathered at an estimated 2,600 watch parties nationwide.

“It is absurd, it is beyond belief, that here in America we remain the only major country on Earth not to guarantee health care to all people. If that proposal can win in Colorado, I believe that idea will spread around the country, and Our Revolution is supporting [Amendment 69],” he said.

The endorsement came as a much-needed boost for the Amendment 69 campaign, led by Democratic state Sen. Irene Aguilar, which took a hit last week when leading liberal groups ProgressNow Colorado and NARAL Pro-Choice Colorado came out against it.

“It’s hard to imagine a figure whose support of ColoradoCare is more meaningful than Sen. Sanders,” said ColoradoCareYes spokesman Owen Perkins in a Thursday statement.

Mr. Sanders easily defeated former Secretary of State Hillary Clinton in the Colorado Democratic caucuses with 59 percent of the vote to her 40 percent, although Mrs. Clinton ultimately won the party’s presidential nomination.

“He mobilized hundreds of thousands of Coloradans to become politically active, and we look forward to working with Sen. Sanders and his supporters to make history together by enacting a central plank in his presidential platform and a founding feature in the ongoing revolution he has inspired,” Mr. Perkins said.

The measure has created headaches for Democrats hoping to retain left-wing Sanders voters without losing the political center.

The cost of the proposal, which would replace the state’s Obamacare exchange, is estimated $25 billion per year, paid for with a 10 percent tax hike that would give Colorado the highest state taxes in the nation.

The health care industry has raised more than $3.6 million to defeat Amendment 69, a figure that could easily double if the measure becomes a cause celebre for Sanders voters.

The worry for Democrats is that the funding will also have the effect of boosting Republicans, who have already moved to tie the measure to Democratic candidates.

Even before the Sanders blessing, ColoradoCare had exposed a rift in the party. Democratic Gov. John Hickenlooper and Sen. Michael Bennet have come out against the measure, while the rank-and-file added support for ColoradoCare to this year’s state Democratic platform.

Jonathan Lockwood, executive director of the free-market group Advancing Colorado, unloaded Friday on Our Revolution, which he described as a “dark money organization.”

“ColoradoCare will restrict women’s access to health care choices, burden seniors, veterans and millennials with an unfair tax scheme, triple taxes on low-income workers and ration much-needed health care,” said Mr. Lockwood.

Supporters of ColoradoCare argue that the measure will actually save residents $4.5 billion per year by eliminating co-pays and deductibles.

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