I need some help trying to understand how annotations work on plasmids such as the one below. I do not understand the double layers structure, for example the light blue "ORF 2" has a annotation layer on top which is red for the GOI "firefly_Luciferase".
How can there be a ORF where a gene has already been inserted? Or does this mean that the gene can have a site-directed mutagenesis or be replaced by another GOI? Or is it just a double strand...