Plasmid_Backbone

Part:BBa_K2941004

Designed by: Pyry Niemel   Group: iGEM19_GemTurku   (2019-10-16)


Vector for periplasmic expression of Fab fragments in E. coli

This vector vector contains Sfi restriction sites to easily clone and express Fab fragments with various specifities to antigens. It is meant to be used with with BBa_K2941002 and its derivatives.

The main function of this vector is to confer the required properties to any Fab protein it is used to produce. First of all it contains a SfiI restriction site inside a PelB signal sequence. This signal sequence is used to relocate the produced protein to periplasmic space. Expression can be induced with IPTG as the vector contains Lac promoter upstream of the PelB .

Other parts of this plasmid contain ampicillin selection marker and ColE1 replication origin.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3355
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 3361
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 3355
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 3355
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 3355
    Plasmid lacks a suffix.
    Illegal XbaI site found at 3370
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 1056


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Categories
Parameters
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