Part:BBa_K5198101

Igk_SEAP

IgK-SEAP is the Secreted Embryonic Alkaline Phosphatase (BBa_K1470004 ) with the Ig Kappa tag (BBa_K4646003 ).

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. When attached to the Ig Kappa tag, this IgK-SEAP signal peptide can be used in solubility studies to find the optimal secretion signal peptide for neoantigens in ATLAS (Figure 1).

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 254
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 565
  Illegal NgoMIV site found at 1534
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 895
  Illegal BsaI.rc site found at 1384