Tityus serrulatus venom is composed by several components especially neurotoxins that acts on ion channels (Na+, K+ or Ca2+) and are responsible for neurotoxic effects on envenoming. These neurotoxins have been widely used in the study of ion channels, as well as in treatment of diseases related to them. Only small amounts of toxins can be obtained from scorpion venom. Therefore, to obtain them in larger quantities it is necessary to perform its cloning and expression in heterologous systems. This study presents the cloning of Ts15, a neurotoxin that act on K+ channels. The Ts15 sequence was obtained through analysis of a cDNA library. Specific primers were designed containing cleavage sites for XhoI/Kex2 and SalI restriction enzymes. The PCR products and the expression vector for secreted proteins pPICZαA (Invitrogen) were digested with the restriction enzyme, applied on 1% agarose gel and purified. The ligation of the vector with the gene was performed by reaction with the enzyme T4 DNA ligase for 16 h at 16 °C. The recombinant plasmid was used for transformation of Escherichia coli DH5α cells by heat shock, plated on LB medium containing 25 mg/mL Zeocin and incubated at 37 °C, for 16 h. The recombinant clones will be subject to DNA sequencing to confirm the insert presence and then to transformation in Pichia pastoris for heterologous expression. This study presents a strategy for cloning Ts15, which will be important to produce the recombinant toxin expressed in heterologous system, for future applications.