Table 4.

Primers and PCR conditions for promoter subcloning.

Construct Prefix (_lcpl :Cre-P2A-EGFP-CAAX)	Forward Primers	Primer Pair Annealing Temp (Ta)*
−822	5’ TGC ATT ACA TAC AGG AAC AGC GG 3’	68 °C
−658	5’ AAA CTG CAT GCT GTG ATT ACT TC 3’	64 °C
−481	5’ AAA CTG ATA TGT GTG AGC TGA CCT 3’	67 °C
−169	5’ TCA TCA CTG AAC ACT GAC TGC A 3’	67 °C
0, ‘Kozak only’	5’ AAA CAT GGC GAA TTT ACT GAC GG 3’	68 °C
Reverse Primer
All constructs	5’ GTC TCT GTA CTG ACC GAC TTT AGT GA 3’

^*We calculated annealing temperatures for these reactions using the manufacturer’s on-line calculator (tmcalculator.neb.com) which is recommended for the specific

polymerase used (Q5 High-Fidelity 2× Master Mix, New England Biolabs).