In the world of plants the non-expresser of pathogenesis-related gene 1 (NPR1) is a key regulator of salicylic acid (SA) mediated systemic acquired resistance (SAR). It also plays an important role in the Jasmonic acid (JA) induced systemic resistance (ISR) signaling pathway and also arbitrates the crosstalk between SA-JA defense pathways to fine-tune defense responses of whole plant system. NPR1 is one of the most agronomically important genes which are under extensive research for development of transgenic with broad spectrum disease resistance. In this study, we have isolated NPR1 gene from genomic DNA of Arabidopsis thaliana ecotype Col-0. 2277 bp AtNPR1 PCR product with single complete open reading frame has been cloned in pJET1.2 cloning vector and sequence characterized. AtNPR1 encode a putative functional protein of 593 amino acids long. Sequence analysis in NCBI CDD domain reveals presence of important Ankyrin repeats domain and a BTB/POZ domain, found in some regulatory proteins and both of which mediate protein-protein interactions. Among the SAR-related gene with vast potential in disease resistance, AtNPR1 is a leading candidate represents promising results in engineering resistance to broad-spectrum of pathogens.