Table 1.

Primer Homologous Sequences for 2A and IRES Elements with Annealing Temperature Adjusted to ~50°C

Genetic Element	Plasmid	5′ primer sequence	3′ primer sequence
T2A	pCAG FlpO-2A-Cre EV	GAGGGCAGGGGAAGT	GGGCCCGGGATTTTCCT
P2A sequence 1	MADR pDonor-H3F3A-K27M-EGFP pTV1 Pdgfra D842V COTv1 Trp53-V5 WPRE	GCCACGAACTTCTCT	AGGACCGGGGTTTTC
P2A sequence 2		GCAACCAATTTTTCACTC	AGGCCCAGGATTCTC
IRES	pEF1a-IRES-NEO	ACGTTACTGGCCGAA	CTTTGAAAAACACGATGATAAT

These sequences can be combined with specific overhangs to design the primers Fw3 and Rv3.