Part:BBa_K642007

Constitutive Adh1 distal-Gal1 proximal promoter tagged with yeGFP

This is the distal region of the mid length Adh1 promoter (BBa_K319005) submitted last year by the uOttawa team fused to the proximal region of the Gal1 promoter. We have demonstrated that it is a functional chimeric promoter and is inducible in the presence of galactose. The promoter is also tagged with the reporter yeGFP (BBa_K319039) which was submitted by the uOttawa team last year.

Characterization of BBa_K642007

We were able to characterize the expression of the Adh1 distal-Gal1 proximal promoter by cloning yeast enhanced GFP downstream of it. The promoter was integrated using NatMX selection into the Ade4 locus of a BY4742 strain of S. cerevisiae. The resulting strain was innoculated in 3 mL of 1x SM + 2% Galactose + 2% Adenine overnight. The overnight culture was reinoculated into 1 mL of the same media at an OD600 of 0.02. After three hours the strain was analyzed for GFP fluorescence on a Cyan ADP flow cytometer.

Figure 1: Histogram of galactose induced wildtype BY4742

Figure 2: Histogram of the galactose induced Adh1 distal-Gal1 proximal promoter with GFP cloned downstream

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 89
  Illegal BsaI.rc site found at 1379