The gut is a tubular, endodermal organ for digesting food and absorbing nutrients. In this study, we characterized eight enhancer detection lines that express green fluorescent protein (GFP) in the whole or part of the digestive tube of the ascidian Ciona intestinalis. Three enhancer detection lines for the pyloric gland, a structure associated with the digestive tube, were also analyzed. These lines are valuable markers for analyzing the mechanisms of development of the gut. Based on the GFP expression of the enhancer detection lines together with morphological characteristics, the digestive tube of Ciona can be subdivided into at least 10 compartments in which different genetic cascades operate. Causal insertion sites of the enhancer detection lines were identified, and the expression pattern of the genes near the insertion sites were characterized by means of wholemount in situ hybridization. We have characterized four and two genes that were specifically or strongly expressed in the digestive tube and pyloric gland, respectively. The present data provide the basic information and useful resources for studying gut formation in Ciona.