Composite

Part:BBa_K5198009

Designed by: Robert Kaptur   Group: iGEM24_Duke   (2024-08-22)
Revision as of 01:59, 30 September 2024 by Rk320 (Talk | contribs)


Gal4-5xUAS-miniCMV-CD19ecto


The ATLAS expression cassette for the extracellular secretion of CD19ecto antigen. Gal4-VP64-inducible expression, either via the synNotch system or a positive control activator system. This part is composed of 5 repeats of Gal4 Upstream Activating Sequence (UAS) (BBa_K4839011), the minimal version of the CMV promoter (BBa_K5198006), and the CD19ecto extracellular domain (BBa_K5198000). The expression of the antigen (or any payload) can be induced by activated synNotch receptors fused to Gal4-VP64 [1]. The CD19ecto contains its native signal peptide, which allows for extracellular export of the protein.

This construct is a key component of Duke iGEM's antigen amplifier circuit. The system is activated when the antiCD19-synNotch receptor recognizes surface-bound CD19, triggering the release of the Gal4-VP64 transcription factor. Gal4-VP64 then translocates to the nucleus, binds to the Gal4 UAS sequence of this part, driving the increased expression of the CD19 antigen ectodomain. The secreted CD19ecto can dimerize and activate nearby CAR-T cells, amplifying the therapeutic response [2].


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 469
    Illegal SapI.rc site found at 305


[edit]
Categories
Parameters
None