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. This sequence was taken from pRS401 plasmid [4].
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
- [4]- pRS401 plasmid
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 649
Illegal XbaI site found at 378 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 649
- 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 649
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 649
Illegal XbaI site found at 378 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 649
Illegal XbaI site found at 378
Illegal NgoMIV site found at 268 - 1000COMPATIBLE WITH RFC[1000]
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