The temporary cycle lane on Bassett Avenue between Winchester Road roundabout and Chilworth Road roundabout will be removed next week.
The move means that between these points Bassett Avenue will revert back to two lanes of traffic in each direction.
The news has been confirmed by Southampton City Council and work will be undertaken overnight on September 2 and 3.
Civic chiefs said the decision follows “a steady increase in traffic”.
But opposition bosses said the scheme should have never been in place.
It comes as concerns were raised over the measures and the council’s Green Transport Recovery Plan was “called back in” to be scrutinised further.
The city council described the pop-up cycle lane on Bassett Avenue as “a positive trial”.
The authority said additional segregation have been introduced between Winn Road and Winchester Road and the temporary cycle lanes in this section will remain.
The 30mph speed limit will also remain in place and buses will be allowed to turn right into Burgess Road from The Avenue.
Other pop-up schemes on Hill Lane, Bitterne Road West, Dale Road and Portswood Road will remain for now and continue to be monitored,the council said.
Southampton Itchen MP Royston Smith, below, said the money should have been spent differently.
He said “it is just a matter of time” before other pop-up cycle lanes will be removed.
He added: “Even they can’t be that foolish as to believe that the city won’t be gridlocked as a result of bus and cycle lanes.
“I don’t think any of them should have been put in place in the first instance.
“These have nothing to do with social distancing.”
Cllr Steve Leggett, cabinet member for green city and place, said the pop-up cycle lane was part of the council’s transport plan.
He added: “This plan is a response to Government policy and funding, which has enabled us to put measures in place that support social distancing following the pandemic.
“We have been closely monitoring the scheme throughout, drawing on a significant level of data to review how the changes have performed at every stage.
Cllr Steve Leggett
“With this data now showing an increase in traffic and some journey times, this is a sensible time to now withdraw the scheme.
“This has been a positive trial, with the measures working well in the early quieter periods.
“We have seen significant numbers of people using the temporary cycle lanes on Bassett Avenue , with highs of over 400 people a day.
“This speaks volumes .”
Work to remove the cycle lane will be carried out between 8pm and 6am. The road will be closed .
Other measures are continued to be monitored. To have your say email firstname.lastname@example.org.