The AA say other sections of the route could now become bottlenecks
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Motorists are not yet able to use part of a new £38.5m underpass because there are no road markings.
While northbound traffic has been using the road to avoid the A34 bottleneck since Thursday, white lines are still to be painted on the southbound route.
The Highways Agency says the work depends on weather but they hope to have it completed by Monday.
The A34/M4 junction, near Chieveley, Berkshire, is described as one of the UK's busiest and most congested.
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For too long the area has suffered chronic congestion, affecting businesses, the environment and quality of life for hundreds of thousands of people
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Linking the South Coast with the Midlands, it carries an average of 60,000 vehicles a day.
A government spokesman said they had now removed the last remaining bottleneck on the route.
The AA says during the construction work motorists have had to endure daily tailbacks of up to seven miles.
Though welcoming the new road, the motoring organisation say it fears it could now show up other bottlenecks.
Paul Watters, of the AA Trust, said: "Life at Chieveley now looks set to be much easier for motorists, hauliers and local residents.
"For too long the area has suffered chronic congestion, affecting businesses, the environment and quality of life for hundreds of thousands of people."
The trust have now called for the government to look at the A34/M3 interchange at Winnall, near Winchester.