Part:BBa_K1907003

CTT1 promoter for S. cerevisiae

Introduction

CTT1 is a gene for cytosolic catalase T. This protein has an important role in protecting the cell from oxidative damage caused by hydrogen peroxide. As Ctt1p is produced under oxidative stress, its promoter is activated in these conditions. (Saccharomyces genome database ID: S000003320)

This part contains a promoter region (924 bp) of CTT1 gene. The length of promoter region is chosen so that contains identified Skn7 and Yap1 binding sites and doesn’t overlap with the next gene in the genome. (He et al., 2005) Skn7 and Yap1 are the main transcription factors activated in oxidative stress and are thus responsible for CTT1 promoter activation. In the CTT1 promoter region, these binding sites are rather close to start codon (-147 bp), and as there are no genes close to it, a longer promoter was decided to be used just in case. The gene sequence for the CTT1 promoter region is obtained from strain S288C of Saccharomyces cerevisiae from the Saccharomyces Genome Database.

The functionality of this promoter was tested in association with the Venus YFP reporter in S. cerevisiae strain SS328-leu, with oxidative stress being induced by hydrogen peroxide. Hydrogen peroxide didn’t seem to activate the promoter, but reaching higher cell densities in cell growth did. This promoter could thus be e.g. used in monitoring the growth phase of yeast cells.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 402
  Illegal PstI site found at 695
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 402
  Illegal NheI site found at 701
  Illegal PstI site found at 695
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 402
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 402
  Illegal PstI site found at 695
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 402
  Illegal PstI site found at 695
  Illegal AgeI site found at 555
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 508
  

Please find the composite part of CTT1 promoter part and Venus YFP part, with which we studied the behaviour of our promoter from here: https://parts.igem.org/Part:BBa_K1907009

References

He, X.J. and Fassler, J.S., 2005. Identification of novel Yap1p and Skn7p binding sites involved in the oxidative stress response of Saccharomyces cerevisiae. Molecular microbiology, 58(5), pp.1454-1467.