Composite

Part:BBa_K5198100

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


HA-SEAP
SEAP acts as a continuous quantitative reporter for gene expression by repeatedly assessing the activity of certain promoter/enhancer elements in many cell samples. SEAP is contained in exosomes and is normally secreted into the extracellular space. 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. HA-SEAP is a SEAP (BBa_K1470004) with an HA tag (BBa_K1150016). The part can be used as a negative control in secretion studies as HA tag does not signal extracellular export of the protein (Figure 1).

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 236
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 547
    Illegal NgoMIV site found at 1516
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 877
    Illegal BsaI.rc site found at 1366


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Parameters
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