Plasmid_Backbone

Part:BBa_K1597001

Designed by: Marieke Mulder   Group: iGEM15_Groningen   (2015-09-09)
Revision as of 10:34, 15 September 2015 by Marieke.groningen (Talk | contribs)

ThrC integration backbone for B. subtilis

Replication of this BioBrick can via integration in the ThrC locus from Bacillus subtilis .

During Bacillus subtilis transformation, the wanted DNA gets integrated in a loci present in the genomic DNA. One of these loci is the ThrC locus. This backbone is based on the plasmids pdg1664 and K823023. The AmyE loci from K823023 were replaced with the ThrC loci from pdg1664. Along with this exchange, the chloramphenicol resistance gene was replaced with erythromycin resistant gene (wich was also found in the pdg1664). This biobrick can be replicated both in Escherichia coli and in Bacillus subtilis, where it integrates into the genome.

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 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


[edit]
Categories
Parameters
None