Part:BBa_K3600004

GLR1 promoter part plasmid

GLR1 promoter part plasmid was made in order to create a part plasmid which contains the GLR1 promoter and is compatible with the Yeast Toolkit.

To create a promoter part plasmid compatible with the yeast toolkit, at first, we designed a gBlock consisting of the 500bp promoter region, that was retrieved from the yeast genome database, and two linking sequences which were added to the ends of the promoter. Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) through a BsmBI assembly.

Usage and Biology

The GLR1 gene encodes the glutathione reductase, which catalyses the reduction of the oxidized form of glutathione (GSSG) to Glutathione (GSH).¹ The GLR1 promoter is regulated by the transcription factor Yap1p. The binding site for Yap1p, in the promoter GLR1, is TTACTAA.²

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 778
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 778
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 778
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 624
  Illegal BsaI.rc site found at 1145
  

References

[1] Grant, C. M., Collinson, L. P., Roe, J. H. & Dawes, I. W. Yeast glutathione reductase is required for protection against oxidative stress and is a target gene for yAP-1 transcriptional regulation. Mol. Microbiol. 21, 171–179 (1996).

[2] Kuge, S. & Jones, N. YAP1 dependent activation of TRX2 is essential for the response of Saccharomyces cerevisiae to oxidative stress by hydroperoxides. The EMBO Journal 13, 655–664 (1994).