Plasmid_Backbone

Part:BBa_K1597001:Design

Designed by: Marieke Mulder   Group: iGEM15_Groningen   (2015-09-09)


ThrC integration backbone for B. subtilis


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 5126
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 5132
  • 21
    INCOMPATIBLE WITH RFC[21]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal EcoRI site found at 5126
    Illegal BamHI site found at 5108
    Illegal XhoI site found at 2503
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found at 5126
    Illegal suffix found at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found at 5126
    Plasmid lacks a suffix.
    Illegal XbaI site found at 5141
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NgoMIV site found at 2467
    Illegal NgoMIV site found at 3606
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Plasmid lacks a prefix.
    Plasmid lacks a suffix.
    Illegal BsaI.rc site found at 3430


Design Notes

pdg1664 contained an EcoRI and a XbaI restriction site. However these sites were situated around a locus. An option was to introduce the prefix, suffix and remove the other restriction site. Here is chosen to copy the loci and erythormycin resistance gene and use these to replace the AmyE intergration loci and chloremphencicol resistance gene from Bba_K823023.


Source

This backbone comes from combining Bba_K823023 and pdg1664

References