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Kent County Council says the idea of a hub airport in the Thames Estuary is the wrong way to address the growing demand for more aviation capacity.
A blueprint that sets out the authority's solutions for meeting the rising demand says there should instead be a fast rail link between Heathrow and Gatwick.
It also argues there should be a new runway at Gatwick once the block on expansion there comes to an end.
Council chiefs also call for greater use of other airports in Kent and the south east, including Manston and Lydd.
The council's formal opposition to the idea of Boris Island or other schemes - including from renowned architect Lord Foster - that have been floated for north Kent pitches the Conservative-run county council against the recently re-elected Tory London Mayor.
Boris Johnson has already indicated he intends to pursue his call for a new hub airport in his second term, arguing the government should seize the nettle and calling for an "act of bravery".
But KCC says that is the wrong solution, saying it would take too long to build, be too expensive and environmentally damaging.
In Bold Steps for Aviation, the policy document released this week, it states: "No action is not an option but action to address capacity issues must be taken quickly; rather than depending on an estuary airport that will take years to develop and may not even succeed, better use of our existing hub and regional airports now will ensure that the UK retains its premier position as a hub airport."
In a clear sign of that KCC intends to fight the mayor over the scheme, Cllr Bryan Sweetland (Con), cabinet member for transport, said: "We should remember he's the mayor of London, not the Prime Minister or leader of KCC.
"We believe there is capacity in our current airports."
On Manston, the county council states it could "strongly complement Heathrow and Gatwick as they increasingly focus on accommodating long-haul flights at the expense of domestic and near-European services".
It adds that its development as a regional airport would create jobs in what remains the county's economic blackspot, saying as many as 6,000 extra jobs could be created for the local economy.
But it accepts transport links will need to be improved if the airport is to "truly succeed" as a regional airport, saying that new a Thanet Parkway station is needed.