In smart cities, smart transportation (ST) is one of the critical infrastructures for efficient traffic routing, congestion-free roads, and for development of sustainable transportation. The advancement of vehicular technologies, Internet of Things (IoT), high-speed communication channel, allows ST to become a major application of IoT system and an important area for research. Unlike traditional transport systems, ST demands secure communication and rapid validation for ensuring data integrity. In this context, a 64-bit Physical Unclonable Function (PUF) was developed for use in the ST. In this work, PUF shows 94.14% reliability, 34.02% uniqueness, and the time required to generate key is 95.8ms.