Part:BBa_K3600003

GSH1 promoter part plasmid

GSH1 promoter part plasmid was made in order to create a part plasmid which contains the GSH1 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.These linking sequences contained a BsaI and a BsmBI cutting site each to make Golden Gate assembly possible.Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) through a BsmBI assembly.

Usage and Biology

The GSH1 gene encodes gamma-glutamylcysteine synthetase.¹ The GSH1 promoter is regulated by the transcription factor Yap1p. The binding site for Yap1p, in the promoter GSH1, is TTACTAA.¹. Using the yeast toolkit for modular assembly, the GSH1 promoter plasmid was used as a part of type II.

Sequence and Features

Assembly Compatibility:

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

  Illegal XhoI site found at 975
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 563
  Illegal BsaI site found at 972
  Illegal BsaI.rc site found at 1084
  

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References

[1] Wu, A. L. & Moye-Rowley, W. S. GSH1, which encodes gamma-glutamylcysteine synthetase, is a target gene for yAP-1 transcriptional regulation. Molecular and Cellular Biology 14, 5832–5839 (1994).