Plasmid_Backbone

Part:BBa_K4727001:Design

Designed by: Uni-Padua-IT 2023   Group: iGEM23_Uni-Padua-IT   (2023-10-02)


pPAC plasmid backbone for P. aeruginosa


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Plasmid lacks a prefix.
    Illegal suffix found at 1007
    Illegal EcoRI site found at 987
  • 12
    INCOMPATIBLE WITH RFC[12]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 987
    Illegal SpeI site found at 1008
    Illegal PstI site found at 1022
    Illegal NotI site found at 993
    Illegal NotI site found at 1015
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 987
  • 23
    INCOMPATIBLE WITH RFC[23]
    Plasmid lacks a prefix.
    Illegal suffix found at 1008
    Illegal EcoRI site found at 987
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 987
    Plasmid lacks a suffix.
    Illegal SpeI site found at 1008
    Illegal PstI site found at 1022
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI site found at 1198
    Illegal SapI.rc site found at 767


Design Notes

This plasmid was assembled using the Gibson assembly method, with fragments coming from three different plasmids: the backbone, including Ori for P. aeruginosa and E. coli, comes from pUCP20 PilB; the terminators region comes from pMF230; the chloramphenicol acetyl-transferase cassette comes from pSB4C5. Through PCR mutagenesis the BioBrick RFC[10] prefix and suffix were added.


Source

Derived from three plasmids: pUCP20 PilB (Addgene Plasmid #162845), pMF230 (Addgene Plasmid #62546) and the standard backbone pSB4C5

References