The available promoters in the Pichia pastoris expression platform are still limited. We selected and identified a novel strong constitutive promoter, P (GCW14) , and tested its promoter activity using enhanced green fluorescent protein (EGFP) as a reporter. Potential promoter regions of P (GCW14) were cloned upstream of the EGFP gene and promoter activity was analyzed by measuring fluorescence intensity. P (GCW14) exhibited significantly stronger promoter activity than the classic strong constitutive promoters P (TEF1) and P (GAP) under various carbon sources, suggesting that P (GCW14) is a strong and constitutive promoter. Hence, P (GCW14) can be used as a promoter for high-level expression of heterologous proteins.