Part:BBa_K3570018

MET17 selection marker

Usage

The MET17 gene, found in the Saccharomyces cerevisiae yeast, encodes a bifunctional enzyme with O-acetylserine and O-acetylhomoserine sulfhydrylase activities involved in methionine and cysteine metabolism. It catalyzes the reaction between acetylated serine or homoserine with thiol to produce the corresponding amino acid and is localized to the cytoplasm and plasma membrane[3].

MET17 gene serves as a commonly used yeast selectable marker. When MET17 gene is inserted into an integrative or replicative plasmid, MET17 allows to counter-select the cells that acquired the prototroph character for methionine so that they can grow without methionine addition in the medium. Those cells should not have the functional MET17 gene in its genome[1].

The sequence contains MET17 specific promoter, MET17 coding sequence, and MET17 terminator.

Experiments

Team iGEM Toulouse 2020 did not have sufficient time to complete the cloning and hence, to test this part functionality.

References

• [1]- Old, R. W., & Primrose, S. B. (1981). Principles of gene manipulation: an introduction to genetic engineering (Vol. 2). Univ of California Press.
• [2]- SGD:S000004294

• [3]- Masselot, M., & de Robichon-Szulmajster, H. (1975). Methionine biosynthesis in Saccharomyces cerevisiae. Molecular and General Genetics MGG, 139(2), 121–132. https://doi.org/10.1007/bf00264692

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 649
  Illegal XbaI site found at 378
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 649
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 649
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 649
  Illegal XbaI site found at 378
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 649
  Illegal XbaI site found at 378
  Illegal NgoMIV site found at 268
  
• 1000
  COMPATIBLE WITH RFC[1000]