Part:BBa_K5198111

nSEAP-sfGFP

nSEAP-sfGFP is the sfGFP (superfolder GFP) (BBa_K5198012 ) with native SEAP tag (BBa_K5198010). Native SEAP tag is the native signal sequence of secreted embryonic alkaline phosphatase (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 secreted into the extracellular space. nSEAP tag sequence causes extacellular secretion of proteins. sfGFP is a robustly folded version of GFP. Its fluorescence is directly proportional to total fusion protein expression. When attached to the nSEAP tag, this nSEAP-sfGFP signal peptide can be used in solubility studies to find the optimal secretion signal peptide for neoantigens in ATLAS (Figure 1).

Figure 1. Fluorescence readings of sfGFP with different signal peptide fusions in HEK293T cell supernatant. The negative control (NC) is sfGFP fused with an HA tag, which does not exhibit 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
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 624
  
• 1000
  COMPATIBLE WITH RFC[1000]