Composite

Part:BBa_K5198102

Designed by: Robert Kaptur   Group: iGEM24_Duke   (2024-08-23)


IFN_SEAP

IFN-SEAP is the Secreted Embryonic Alkaline Phosphatase (BBa_K1470004) with the Interferon tag (BBa_K4646001).

SEAP acts as a continuous quantitative reporter for gene expression by repeatedly assessing the activity of certain promoter/enhancer elements in many cell samples. This reporter’s activity can be assayed quicker, more easily, and more cheaply than other reporter enzymes’ activities. SEAP activity can be detected with conventional chemiluminescent assays. The IFN tag is derived from the interferon alpha protein signal sequence (INF α), a cytokine produced by the innate immune system in response to viral infections and other environmental exposures. When attached to the Interferon tag, this IFN-SEAP signal peptide can be used in solubility studies to find the optimal secretion signal peptide for neoantigens in ATLAS. This signal peptide excretes into the extracellular space.

Description of the image

Figure 1. Chemiluminescent readings of SEAP with different signal peptide fusions in HEK293T cell supernatant. The negative control (NC) is SEAP fused with an HA tag, which does not signal extracellular secretion. Error bars represent the standard deviation (SD) of 4 biological replicates. The data was normalized by subtracting background fluorescence.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 278
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 589
    Illegal NgoMIV site found at 1558
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 919
    Illegal BsaI.rc site found at 1408


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