Phosphoenol pyruvate carboxylase (PEPc), a key enzyme in photosynthesis, an important carboxylase for plant cell metabolism, fixes CO2 in mesophyll cells to form dicarboxylic acids, and being more specific for inorganic carbon, improves the photosynthetic rate of C4 plants. In the present study, partial gene sequencing of PEPc was done using Saccharum spp. hybrid, CoLk 94184 of sugarcane, one of the most popular variety released by ICAR-IISR, Lucknow. Utilizing PEPc of different cultivars of sugarcane species, the primers were designed for PEPc gene and the first ever nucleotide sequence was determined, especially for Saccharum spp. hybrid CoLk 94184 (accession no: MW766370.1). Sequence based phylogenetic tree showed close relatedness between the PEPc sequences for various Saccharum cultivars, thus pointing to little genetic change, while that for sorghum and maize were relatively distantly related. Results may help in understanding the photosynthetic process using sugarcane hybrid specific genes