Jim Sillars would turn down Scottish Independence if it meant rejoining the EU
The Nationalist veteran said he would abstain and warned Nicola Sturgeon many of her own supporters would do the same.
Mr Sillars, who played a leading role in the 2014 Yes campaign, said he did not want to be governed by an “unelected elite”.
He also accused the First Minister of behaving “hysterically” over her threats of another ballot if Scotland leaves the EU single market with the rest of the UK.
The pro-Brexit former MP said making any poll about EU membership would be a “big mistake” and alienate SNP supporters, a third of whom voted Leave last year.
He said: “I, for example, could not vote Yes if on the ballot paper it said, ‘We wish the Scottish state to be a member of the European Union’, and I’m not alone in that.
“One of the biggest miscalculations by Nicola Sturgeon is to believe that the 1.6m Scots who voted Remain would automatically then vote to go back into the European Union.
The former SNP deputy leader has slammed Sturgeon's approach to Brexit
“That means Ruth Davidson, the leader of the Tory party, and all the Tories who voted to Remain, would in fact vote to leave the United Kingdom and take a Scottish state into the European Union. I think that’s fantasy.”
I do not want to be run by an unelected, self-serving elite
Ms Sturgeon has stepped up her threats to break up the UK, arguing another poll would be "almost necessary" if her demands for a bespoke Brexit deal are not met by Prime Minister Theresa May.
She is expected to take further steps towards a rerun of the 2014 ballot at next week's SNP conference in Aberdeen.
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Her predecessor Alex Salmond has repeatedly suggested autumn 2018 as a date.
Mr Sillars' interventon comes after a senior SNP figure suggested Mr Salmond was mistaken during the independence campaign three years ago.
Andrew Wilson, who chairs the SNP's growth commission, rejected the former First Minister's boast that oil revenues would have been a "bonus" rather than a necessity.
Former SNP leader Gordon Wilson has also warned that the party is caught in a “political vortex” and Ms Sturgeon cannot avoid demanding a referendum "at the worst possible time".
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Mr Sillars, who has previously claimed at the time that five or six SNP MSPs backed Leave in defiance of the First Minister, added: “I do not want to be run by an unelected, self-serving elite."
Opponents seized on the his remarks describing them as "stark reminder" not ignore Scots Leave voters.
Scottish Tory MSP Ross Thomson said: “Among those were 400,000 SNP voters, the party’s former deputy leader, and as many as six SNP MSPs.
Nicola Sturgeon has been pushing for a second Scottish Independence referendum
“The First Minister should remember that next time she embarks on another Brexit rant."
Scottish Lib Dem leader Willie Rennie added: "Jim Sillars speaks for thousands of independence supporters who backed Brexit.
"The real test for Nicola Sturgeon is whether she will hold true to her pro-European beliefs or sell them out to get Jim Sillars and others back on board for another independence referendum."