There exists an electromagnetic shielding effect on radio signals in a tunnel, which results in no satellite positioning signal in the tunnel scenario. Moreover, because vehicles always drive at a high speed on the highway, the real-time localization system (RTLS) has a bottleneck in a highway scenario. Thus, the navigation and positioning service in tunnel and highway is an important technology difficulty in the construction of a smart transportation system. In this paper, a new technology combined downlink time difference of arrival (DL-TDoA) is proposed to realize precise and automated RTLS in tunnel and highway scenarios. The DL-TDoA inherits ultra-wideband (UWB) technology to measure the time difference of radio signal propagation between the location tag and four different location base stations, to obtain the distance differences between the location tag and four groups of location base stations. The proposed solution achieves a higher positioning efficiency and positioning capacity to achieve dynamic RTLS. The DL-TDoA technology based on UWB has several advantages in precise positioning and navigation, such as positioning accuracy, security, anti-interference, and power consumption. In the final experiments on both static and dynamic tests, DL-TDoA represents high accuracy and the mean errors of 11.96 cm, 37.11 cm, 50.06 cm, and 87.03 cm in the scenarios of static tests and 30 km/h, 60 km/h, and 80 km/h in dynamic tests, respectively, which satisfy the requirements of RTLS.