Protein_Domain

Part:BBa_K5109004

Designed by: Veronika Rofner   Group: iGEM24_Uni-Padua-IT   (2024-09-25)


Lpp-OmpT Carrier with Linker

The basic part BBa_K5109003 was constructed by adding a linker sequence to a fragment encoding a carrier protein. As described in the part concerning the entire display system, the linker is positioned downstream of the carrier and upstream of the passenger protein. The purpose of the linker is to increase flexibility in the display system, providing the anchored enzyme (passenger) with more space to perform its function. To link the passenger to the carrier, we selected a sequence that coincides with a thrombin cleavage site. This allows for controlled cleavage: by supplying thrombin to the medium in which the cells are cultured, the linker can be cleaved, enabling the isolation of the surface-expressed protein. Additionally, NdeI restriction sites were introduced at both ends of the basic part. This facilitates its insertion into an expression cassette, such as BBa_K5109001, which contains an NdeI cut site.

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 549
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None