The pM36 plasmid consists of a recombination-mediated cassette exchange (RMCE) cassette flanked by a pair of inverted attP sites. This cassette contains a mutagenic gene trap element consisting of a splice acceptor site, stop codons in all three reading frames and an SV40 polyadenylation signal. A Avic\GFP3xP3.cLa marker allele followed by a polyadenylation signal is present downstream of the gene trap element. pM36 is used to generate donor plasmids that can be used in combination with CRISPR/Cas9 to insert the attP-flanked RMCE cassette into a defined location in the genome by homology directed repair; the donor plasmid is generated by adding homology arms to each side of the pM36 RMCE cassette using the Type IIS restriction sites in plasmid.
The pM36 vector has been found to be unstable in bacteria, frequently recombining out the Avic\GFP3xP3.cLa marker allele (this appears to be due to the 3xP3 promoter fragment in this plasmid being 290bp longer than other vectors that use the same marker).