The Sunday market will be rehomed after 27 years trading at Brighton Station.
Easter Sunday will be the last time the historic market and tourist attraction will be hosted next to the city's mainline station.
Instead it will be shunted more than three miles away to the Brighton Marina car park.
Organisers said they were forced to up sticks after repeated cancellations caused by rail replacement services.
Last month only one market went ahead as planned - leaving traders thousands of pounds out of pocket.
Brian Nunan, manager of market owner Bray Associates, said: “We're part of the Brighton cultural scene.
“It's survived for 27 years and it's become part and parcel of Brighton life.
“Hopefully we will get a few more people who come from that side of Brighton.
“I decided to look for another place because I was getting fed up with all the cancellations.”
Work to the tracks has meant Sunday train services out of Brighton station have been cancelled and buses used to ferry passengers out of the city.
They depart from an area next to the market and Southern Railway, which owns the land, claim buses were held-up by market traffic.
Councillor Dee Simpson, who has run a clothing store at the market for 25 years, said Easter Sunday would a “sad day”.
She added: “I don't know what will happen.
“It might be OK I always keep an open mind.
“Sunday will be a sad day, we'll have to crack open a bottle. It's been a way of life for me.”
Coun Simpson criticised Southern for not doing more to manage traffic.
She said:“Why couldn't the traffic have been managed? We could have had a one-way system operating.”
The market will run on an initial three-month probationary period at the Marina starting from April 19.
It currently closes at 2pm but, from next Sunday, will have to shut at midday.
Fees will remain at £12 per pitch.
One regular market-goer, Chris Cooke, 35, of Adelaide Crescent in Hove, said he was scared that the move may be a step too far for some punters.
He added: “It will kill it.
“It's too far to travel for most people. It's such a part of Brighton and particularly that area. It's whole character will change.”
But Kirsty Harris, events and marketing manager at Brighton Marina, said she hoped to see the Marina, which has hosted the Mad Hatter market for the past two months, as the city’s “market destination.”
She said: “I believe it will add to our retail opportunities and add another attraction to the Marina.
“We're responding to the current market climate.
“There will still be plenty of parking. Sunday morning is a quiet time but the market will have to shut at midday so we can have the car park back.”
A spokesman for Southern said engineering work meant it was impossible to run buses and the market at the same time.
He added that the company’s first responsibility was to its passengers and that there were no plans to replace the market.
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