South Korean workers were being allowed to leave Kaesong Industrial Complex but not cross into it from the South, Seoul’s Unification Ministry confirmed.
The Kaesong zone at North-South border, which is a money-maker for North Korea, is seen as a key barometer of inter-Korean relations.
The move came as the US called North Korea’s recent rhetoric unacceptable.
Kaesong Industrial Complex is home to more than 100 factories. More than 50,000 North Koreans work there, as well as several hundred South Korean managers.
Permission is granted on a daily basis for workers to cross into the complex, where they can stay overnight. More than 850 South Koreans were at Kaesong when the ban was announced.
“South Korea’s government deeply regrets the entry ban and urges it be lifted immediately,” Unification Ministry spokesman Kim Hyung-seok told reporters.
“Ensuring the safety of our citizens is our top priority and the South Korean government will take necessary measures based on this principle,” he said.
The entry ban is not unprecedented – South Koreans were briefly denied access in March 2009 because of US-South Korea military exercises.
North Korea, which has been angered by UN sanctions imposed after its recent nuclear test and annual US-South Korea military drills, threatened to shut down the complex last week.
In recent weeks North Korea has also threatened attacks on US military bases in Asia and South Korean border islands.
On April 2, North Korea said it planned to restart its mothballed reactor at Yongbyon – the source of plutonium for its nuclear weapons programme.
Later in the day, US Secretary of State John Kerry called recent North Korean actions “dangerous” and “reckless”.
“Let me be perfectly clear here today. The United States will defend and protect ourselves and our treaty ally, the Republic of Korea (South Korea),” John Kerry said after talks with his South Korean counterpart Yun Byung-se.
In recent days the US has responded to North Korea with a series of high-profile flights of advanced aircraft, including stealth fighters and nuclear-capable B-52 bombers, over South Korea.
Officials have also confirmed that the USS John McCain, an Aegis-class destroyer capable of intercepting missiles, has been positioned off the Korean peninsula.
A second destroyer, the USS Decatur, has been sent to the region.
Earlier on Tuesday UN Secretary-General Ban Ki-moon had warned that the crisis had “gone too far” and called for urgent talks with the North.
“Things must begin to calm down, there is no need for the DPRK [North Korea] to be on a collision course with the international community. Nuclear threats are not a game,” Ban Ki-moon said.
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