Part:BBa_K3600001

TRX2 promoter part plasmid

TRX2 promoter part plasmid was made in order to create a part plasmid which contains the TRX2 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

TRX2 gene encodes thioredoxin protein in Saccharomyces cerevisiae.¹ The TRX2 promoter is regulated by the transcription factor Yap1p. The binding site for Yap1p, in the promoter TRX2, is TTACTAA.¹ Using the yeast toolkit for modular assembly, ² the HSP12 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
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 568
  Illegal BsaI.rc site found at 1089
  

References

[1] 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).

[2] Martínez-Pastor, M. T. et al. The Saccharomyces cerevisiae zinc finger proteins Msn2p and Msn4p are required for transcriptional induction through the stress response element (STRE). The EMBO Journal 15, 2227–2235 (1996).