Abstract: Four metallothionein (MT) isoforms have previously been identified in the brine shrimp Artemia. The goals of this study were to isolate at least one of the MT genes, clone a MT promoter region, and characterize the cis-acting sequences that regulate MT gene expression. PCR of Artemia genomic DNA amplified a product containing a MT gene with no internal introns. Three different approaches were made to clone the MT promoter. Firstly, conventional cloning of size-selected genomic DNA into a dephosphorylated vector produced empty clones or no clones. The use of a novel linear cloning vector produced many recombinant clones, but screening of the DNA library did not identify a MT promoter sequence. Lastly, genome walking by PCR amplified a sequence that lacked a TATA box and metal response elements. A blast search identified the sequence as a mitochondrial hydroxyacylglutathione hydrolase found in marine organisms.