The complete mitochondrial genome of an orb-weaving spider Neoscona theisi (Walckenaer) is determined in this article. It is a typical circular duplex DNA molecule with a length of 14,156 bp, which encodes the same 37 genes as all metazoan mitogenomes and an A + T-rich region (D-loop). The overall A + T content is 75.2% (A: 35.5%; T: 39.8%; G: 15.6%; C: 9.3%). All of the protein-coding genes were initiated by ATN, with the exceptions in three genes. COI has a TTA start codon, COII, COIII and ND6 use TTG as initiation codon. Three genes (ND3, COII and ND5) end with incomplete stop codons (T or TA), while all other genes terminated with canonical stop codons (TAA or TAG). Among 22 transfer RNAs genes, six of them (tRNA Ala , tRNA Ser(AGN) , tRNA Ser(UCN) , tRNA Gly , tRNA Arg and tRNA His ) lack the potential to form the cloverleaf-shaped secondary structure. The A + T-rich region is 559 bp with an A + T content of 79.6%.