Difference between revisions of "Part:BBa K3600004"
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<partinfo>BBa_K3600004 short</partinfo>
 
<partinfo>BBa_K3600004 short</partinfo>
  
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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.
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GLR1 promoter part plasmid was made in order to create a part plasmid which contains the GLR1 promoter and is compatible with the Yeast Toolkit.
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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.
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To create a promoter part plasmid compatible with the yeast toolkit, at first, we designed a gBlock consisting of the 500bp promoter region, that was retrieved from the yeast genome database, and two linking sequences which were added to the ends of the promoter. Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) through a BsmBI assembly.
  
 
===Usage and Biology===
 
===Usage and Biology===

Revision as of 15:49, 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 is compatible with the Yeast Toolkit.

To create a promoter part plasmid compatible with the yeast toolkit, at first, we designed a gBlock consisting of the 500bp promoter region, that was retrieved from the yeast genome database, and two linking sequences which were added to the ends of the promoter. Then, the gBlock was inserted into the part plasmid entry vector (pYTK001) 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).