Part:BBa_K2539500

PalcA-Regulated GFP Expression Construct

This construct uses the inducible promoter PalcA (BBa_K2092002). PalcA is activated in the presence of both ethanol and a transcription factor, alcR (BBa_K2092001), and was originally isolated from the fungus Aspergillus nidulans (Panozzo et al., 1997). GFP is used as a visible reporter so we can characterize the function of the promoter PalcA.

Construct Design

The PalcA promoter is placed in front of a strong RBS (BBa_B0034), GFP (BBa_E0040), and double terminator (BBa_B0015). This places the expression of GFP under the control of the PalcA promoter.

PCR Check Results

The part was confirmed by PCR using the primers VF2 and VR, as well as sequencing by Tri-I Biotech.

PCR check for BBa_K2539500 using VF2 and VR primers. Using these primers, PCR produced a band at the expected size of 2 kb.

References

Panozzo C, Capuano V, Fillinger S, Felenbok B. (1997). The zinc binuclear cluster activator AlcR is able to bind to single sites but requires multiple repeated sites for synergistic activation of the alcA gene in Aspergillus nidulans. J Biol Chem. 5;272(36):22859-65.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 274
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1513