Composite

Part:BBa_K5317018:Design

Designed by: Lisa Marie de Sousa Miranda   Group: iGEM24_Hannover   (2024-09-15)


CMV-EGFP-PknB


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 2596
    Illegal SpeI site found at 3259
    Illegal PstI site found at 2021
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal SpeI site found at 3259
    Illegal PstI site found at 2021
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2271
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 2596
    Illegal SpeI site found at 3259
    Illegal PstI site found at 2021
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 2596
    Illegal SpeI site found at 3259
    Illegal PstI site found at 2021
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

Codon-optimized for the expression of pknB in mammalian cells.


Source

To create the composite part, the codon-optimized pknB was integrated into the EGFP-C2 backbone downstream of the EGFP sequence after SalHI and BamHI restriction enzyme digestion. The construct is driven by a constitutively active CMV promoter, resulting in expression of pknB-EGFP. This design allows fluorescent visualization of PknB in mammalian cells.

References