<p style="text-align: justify;">Cotton event COT67B was produced via <i>Agrobacterium</i>-mediated transformation of the Coker 312 cultivar using two plasmid vectors, pNOV4641 and pNOV1914. The T-DNA region of pNOV4641 contained a single copy of the <i>flcry1Ab</i> gene derived from <i>B. thuringiensis</i> subsp. <i>kurstaki</i> strain HD-1. The T-DNA region of pNOV1914 contained a single copy of the selectable marker gene <i>aph4</i>, which encodes hygromycin-B phosphotransferase. Expression of hygromycin-B phosphotransferase confers resistance to the antibiotic hygromycin, which was used for selection of transformed tissue during the initial tissue culture phase of transformation. However, once transformants were identified, traditional breeding was used to identify segregants containing <i>flcry1Ab</i> but lacking the <i>aph4</i> gene, including elements from its plasmid backbone. The <i>aph4</i> gene and related regulatory sequences are no longer present in cotton event COT67B.</p>
