Part:BBa_K4220100

HSF1up-HIS3+NatR+ADH1+CUP1-HSF ORF

Introduction

This is a kill switch in yeast which can let the yeast survive only in the presence of copper ion.

This part is an improvement of part BBa_K2086002.

Why is this an improvement?

We have improved a kill switch in E. coli (composite part) into one which can function in the yeast by replacing each basic parts into ones which have similar functions in the yeast.

Improvements:

• promoter: PyeaR promoter (responsive to nitrate, nitrite and nitric oxide) to CUP1 (responsive to copper ion)
• essential gene: SerA (E. coli) to HSF1 (yeast)

Design

• How to kill the yeast?
  • HSF1: essential gene in yeast [1]
• How to determine when to kill?
  • Copper ion: abundant in the intestine; scarce in the environment [2]
• Others
  • HIS3-NatR-ADH1: resistant gene expression for screening

Results

CUP1p-HSF1: kill switch

YPD: Yeast Extract Peptone Dextrose Medium

Sequence and Features

Assembly Compatibility:

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

  Illegal NotI site found at 247
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 367
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 512
  

References

1 Kopczynski, J. B., Raff, A. C. & Bonner, J. J. Translational readthrough at nonsense mutations in the HSF1 gene of Saccharomyces cerevisiae. Mol Gen Genet 234, 369-378, doi:10.1007/BF00538696 (1992).

2 Nose, Y., Kim, B. E. & Thiele, D. J. Ctr1 drives intestinal copper absorption and is essential for growth, iron metabolism, and neonatal cardiac function. Cell metabolism 4, 235-244, doi:10.1016/j.cmet.2006.08.009 (2006).