Coding

Part:BBa_K3570013

Designed by: Anton Mykhailiuk   Group: iGEM20_Toulouse_INSA-UPS   (2020-10-01)


URA3 selection marker


Usage

The URA3 gene, found in the Saccharomyces cerevisiae yeast, encodes a protein called Orotidine-5'-phosphate (OMP) decarboxylase which catalyzes the sixth step in the de novo biosynthesis of pyrimidines, converting OMP into uridine monophosphate (UMP).

URA3 gene serves as a commonly used yeast selectable marker. When inserted into an integrative or replicative plasmid, URA3 allows to counter-select the cells that acquired prototroph character for uracil so that they can grow without uracil addition in the medium. Those cells should not have the functional HIS3 gene in its genome[1].

The sequence contains URA3 specific promoter, URA3 coding sequence, and URA3 terminator and was taken from FL38 plasmid [2].


Experiments

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

Related parts on regisrty

The BBa_K2407301 part previously described the URA3 gene coding sequence only, which can not be used directly whereas our part contains promoter, coding sequence and terminator.

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]- FL38 plasmid
  • [3]- SGD:S000000747

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 278
    Illegal BglII site found at 1374
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None