Camera enforcement of the new bus gate on Portswood Road will get underway on Monday.
It comes a week after Southampton City Council said the section of road would not be enforced.
It comes after several bumps in the road as hundreds of drivers were seen passing through the restricted area on the first day of the scheme.
But Southampton City Council stands firm by the fact the number of cars using the road has dropped in nine days, and the number of pedestrians has increased.
Councillor Eamonn Keogh, cabinet member for environment and transport said: “We do appreciate that it takes time for people to get used to changes like the ones being trialled in Portswood and so we have allowed two weeks for drivers to adjust to the new restriction without taking any further action.
“From Monday, February 10, we will be issuing warning letters in the first instance to anyone who passes the new bus/taxi/cycle only restriction to ensure people are aware of the restriction and after that we will issue fines if individuals continue to pass the restriction despite having received the warning letter.”
READ MORE: Traders react to restrictions on Portswood Broadway
Between Westridge Road and St Denys spur road, the busy thoroughfare is only open to bus, taxis, and cyclists from 7am to 10am and 4pm to 7pm Monday to Saturday.
This will be in force for a six-month period.
People who drive through the restriction will receive a Warning Notice, and on their second contravention they will receive a Penalty Charge Notice (PCN) of £70, reduced to £35 if paid within 21 days.
All money received from PCNs will be used to pay for the upkeep and running of the camera system and any surplus funds are reinvested into Southampton’s transport network, according to the council.
A PCN will only be issued two weeks after a warning notice is activated.
A spokesperson for the council said: “So far, the monitoring has shown that the number of cars using Portswood Road has fallen since the bus/taxi/cycle only restriction was implemented, compared to the previous week.
“The monitoring has also shown that pedestrian numbers along Portswood Broadway have increased during the first week of the trial compared to the week before.”
Following concerns raised from residents, adjustments will be made to parking on Brookvale Road near the junction with Highfield Lane.
This will help “ease congestion and improve road safety” around the junction, and additional monitoring of Brookvale Road will be carried out as part of the trial process.