UTK Knoxville- October

9/21/12 Colony PCR Confirmed: J63010 +Adh + mOrange + pIRES + GFP + cyc J63010 +Adh + mOrange + pIRES2 + GFP + cyc J63010 +Adh + mOrange + pSAP + GFP + cyc

9/23/12 Colony PCR Confirmed: J63010 + Adh + Neo J63010 +Adh + mOrange + YAP1 + GFP + cyc J63010 +Adh + mOrange + URE2 + GFP + cyc J63010 +Adh + mOrange + HAP4 + GFP + cyc

9/24/12 Digestions: J63010 + pIRES + GFP + cyc : SpeI and PstI J63010 + pIRES2 + GFP + cyc : SpeI and PstI J63010 + pSAP + GFP + cyc : SpeI and PstI J63010 + Adh + Neo : XbaI and PstI J63010 +Adh + mOrange + pIRES + GFP + cyc + Adh + Neo : EcoRI and PstI J63010 + Adh + mOrange +pIRES2 + GFP + cyc + Adh + Neo: EcoRI and PstI J63010 + Adh + mOrange +pSAP + GFP + cyc + Adh + Neo: EcoRI and PstI J63010 + Adh + mOrange + cyc : SpeI and PstI J63010 + Adh + GFP + cyc: SpeI and PstI

9/25/12 Ligations performed to create constructs: J63010 + Adh + mOrange + cyc + Adh + Neo J63010 + Adh + GFP + cyc + Adh + Neo J63010 + pIRES + GFP + cyc + Adh + Neo J63010 + pIRES2 + GFP + cyc + Adh + Neo J63010 + pSAP + GFP + cyc + Adh + Neo

9/28/12 Colony PCR Confirmed: J63010 + Adh + mOrange + cyc + Adh + Neo (AOC) J63010 + Adh + GFP + cyc + Adh + Neo (AGC) J63010 + pIRES + GFP + cyc + Adh + Neo (pIRES control) J63010 + pIRES2 + GFP + cyc + Adh + Neo (pIRES2 control) J63010 + pSAP + GFP + cyc + Adh + Neo (pSAP control)

BY4741 Yeast electroporated with: AOC, AGC, pIRES control, pIRES2 control, pSAP control,pIRES complete, pIRES2 complete, pSAP complete. Electroporations were plated onto 200ug/ml G418 YPD plates.

10/1/12

Colonies finally appeared on plates. Colonies were picked into corresponding 200ug/ml G418 YPD plates. No colonies grew in culture.