Difference between revisions of "Part:BBa K3600004"
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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.
 
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.
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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.
 
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.
  

Revision as of 15:35, 27 October 2020


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).