The Haloacid dehalogenase opens up many potencial application. Gene that codes haloacid dehalogenase of K. pneumoniae ITB1 have amplified by PCR, and cloned into Pgem-t Easy for nucleotide sequencing. Nucleotide sequensing is the first step to analyze functional amino acid sequence of protein. the purpose of this research is to know any mutation in the sequence and find out the restriction site for over expression of this multifunctional enzyme in host.