Composite

Part:BBa_K3425070

Designed by: Tereza Hubackova   Group: iGEM20_UofUppsala   (2020-10-26)


Modular outer membrane display construct for nanobody specific to HSP biomarker

This composite part serves for the display of nanobodies on the outer surface of E. coli. Display of nanobodies on the surface is useful for testing their functionality.

BBa_K3425070 is designed as a modular system of six components. All components are tailored for iGEM Type IIS assembly standard. In contrast to standard iGEM Type IIS assembly, the coding sequence is split into three parts. The middle nanobody module can be easily exchanged for nanobody of different specificity. In this case the nanobody, B-F10 (BBa_K3425043), is specific to the Mycobacterium tuberculosis biomarker HSP16.3 (BBa_K3425041).

Part number Name Description
BBa_J23108 promoter Constitutive promoter of medium strength.
BBa_K3425032 RBS RBS tailored for Type IIS assembly.
BBa_K3425042 pelB Signal peptide directing the fusion protein to the periplasm.
BBa_K3425043 B-F10 Exchangable Module. Nanobody specific for Mycobacterium tuberculosis biomarker HSP 16.3.
BBa_K3425044 ehaA EhaA can be used as an outer membrane display system for E. coli
BBa_B0015 terminator Double terminator. E. coli


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1036
    Illegal AgeI site found at 625
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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