DNA

Part:BBa_K207001

Designed by: Jennifer Jocz   Group: iGEM09_Harvard   (2009-10-21)

PhyB-DBD Fusion for Yeast-Two-Hybrid

This Phytochrome B is sensitive to red light and far red light. Upon being struck by red light it changes conformation and can bind to its interacting factor PIF3. This complex then translocates to the nucleus and induces changes in transcription. In plants this protein is involved in de-etiolation of seedlings.

This fusion protein is for use in a yeast-two-hybrid system, and has a nuclear localization signal and a Gal4 DNA binding domain fused to its C terminus.

Characterization: must be used with PIF3 AD. Yeast transformation optimal DNA amount 1 ug. Optimal cell concentration for experiment 1x10^6. Laser stimulation 660 nm 1 to 30 seconds. [edit]

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 490
    Illegal BamHI site found at 572
    Illegal XhoI site found at 523
    Illegal XhoI site found at 542
    Illegal XhoI site found at 2081
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 2000
    Illegal SapI site found at 739


[edit]
Categories
Parameters
None