Primer

Part:BBa_K268025

Designed by: Nora Yucel   Group: iGEM09_UChicago   (2009-10-21)


YGR243C -> GFP

~2.6kbp linear DNA fragment used for genomic integration and expression expression of GFP under the control of the promoter for YGR243C gene. For transformation in S. cerevisiae only, this part will swap out the YGR243 gene and replace it with a Kan marker+GFP coding region downstream of the YGR243C promoter. Promoter shown to be up-related by the neurotoxin paraoxon and/or hydrolysis of paraoxon to p-nitrophenol and diethyl-phosphate.

This part can be retrieved from the yeast genome in which it was integrated through colony PCR of the following primers: Forward: 62-mer CTGGGAGGAGCTTCAATCGGCGATGCATGGTCAGGCTTTATGcggatccccgggttaattaa Reverse: 60-mer GACATATTTCTTACACAAACGCACCAAAAAAGAAATCTGAgaattcgagctcgtttaaac


Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2461
    Illegal PstI site found at 1225
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2461
    Illegal PstI site found at 1225
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2461
    Illegal BglII site found at 1015
    Illegal BamHI site found at 44
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2461
    Illegal PstI site found at 1225
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2461
    Illegal PstI site found at 1225
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 704


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Categories
Parameters
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