<p dir="ltr"><b>Main conclusion</b> <i>Agrobacterium</i>-mediated transformation of <i>Nicotiana tabacum</i>, using an intragenic T-DNA region derived entirely from the <i>N. tabacum</i> genome, results in the equivalence of micro-translocations within genomes.</p><p dir="ltr"><b>Abstract</b> Intragenic <i>Agrobacterium</i>-mediated gene transfer was achieved in <i>Nicotiana tabacum</i> using a T-DNA composed entirely of <i>N. tabacum</i> DNA, including T-DNA borders and the <i>acetohydroxyacid synthase</i> gene conferring resistance to sulfonylurea herbicides. Genomic analysis of a resulting plant, with single locus inheritance of herbicide resistance, identified a single insertion of the intragenic T-DNA on chromosome 5. The insertion event was composed of three <i>N. tabacum</i> DNA fragments from other chromosomes, as assembled on the T-DNA vector. This validates that intragenic transformation of plants can mimic micro-translocations within genomes, with the absence of foreign DNA.</p>