Primer

Part:BBa_K268024

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


YJL287W -> GFP

~2.6kbp linear DNA fragment used for genomic integration and expression expression of GFP under the control of the promoter for YJL287W gene. For transformation in S. cerevisiae only, this part will swap out the YJL287 gene and replace it with a Kan marker+GFP coding region downstream of the YJL287W 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: 64-mer CAAGATACAAACAAAGCTTTTCAACGTAATATTTACTATCGATGcggatccccgggttaattaa Reverse: 60-mer GGTTTCTTCGCACATTATCATTATTATTCTTTGAGAATACgaattcgagctcgtttaaac

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 2463
    Illegal PstI site found at 1227
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2463
    Illegal PstI site found at 1227
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2463
    Illegal BglII site found at 1017
    Illegal BamHI site found at 46
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 2463
    Illegal PstI site found at 1227
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 2463
    Illegal PstI site found at 1227
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 706


[edit]
Categories
Parameters
None