Plasmid
GLR1pr

Part:BBa_K3600004

Designed by: Sven Spoerri, Yasmine Koubaa   Group: iGEM20_EPFL   (2020-06-14)
Revision as of 15:33, 27 October 2020 by Yasmine koubaa (Talk | contribs)


GLR1 promoter part plasmid

GLR1 promoter part plasmid was made in order to create a part plasmid which contains the GLR1 promoter and compatible with the Yeast Toolkit in the mean time.

At first, a gBlocks consisting of the 500bp promoter region, that was retrieved from the yeast genome database, was added linking sequences to both ends. These linkers contained a BsaI and a BsmBI cutting site each to make Golden Gate assembly possible. Then, the ...was inserted into the part plasmid entry vector (pYTK001) to create GLR1 part plasmid through a BsmBI assembly.


Usage and Biology

  • activator: Yap1p
  • The GLR1 gene encodes glutathione reductase, which catalyses the reduction of the oxidized form of glutathione (GSSG) to Glutathione (GSH).1
  • The promoter GLR1 has been chosen in order to express fluorescent protein [ref] when the S.cervisiae is exposed to oxidative stress.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 778
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 778
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 778
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 624
    Illegal BsaI.rc site found at 1145


Measurements

References

[1] Grant, C. M., Collinson, L. P., Roe, J. H. & Dawes, I. W. Yeast glutathione reductase is required for protection against oxidative stress and is a target gene for yAP-1 transcriptional regulation. Mol. Microbiol. 21, 171–179 (1996).

[edit]
Categories
//chassis/eukaryote/yeast
Parameters
None