Device

Part:BBa_K4646018

Designed by: Alexander Diefes   Group: iGEM23_Duke   (2023-10-10)
Revision as of 07:15, 12 October 2023 by Rk320 (Talk | contribs)

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EF1alpha _Kozak_IL-2

This construct is driven by a strong EF1alpha promoter with enhancer. It also includes a strong Kozak. It can be used to secrete a protein of interest using the IL-2 signal peptide in mammalian systems.

Figure 1: Median YFP fluorescence intensity normalized to median BFP fluorescence intensity in HEK293T cells transfected with secretion constructs. The data was grouped according to promoter that expressed the reporter. HA signal peptide was used as the negative control. Values are mean ± SEM.

The Duke 2023 iGEM team used the signal peptide IL-2 as part of our study to assess the secretion efficiencies of different combinations of signal peptides, promoters, and cargos. Compared to the negative control signal peptide HA, IL-2 showed relatively high levels of secretion as shown by low levels of YFP fluorescence inside of the cells. Based on this study, we found that IL-2 has relatively high secretion efficiency when matched with a strong promoter such as CMV or EF1α.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 643
    Illegal XhoI site found at 1042
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 777
    Illegal AgeI site found at 151
  • 1000
    COMPATIBLE WITH RFC[1000]


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