Composite

Part:BBa_K3445002:Design

Designed by: Madison Hibbs   Group: iGEM20_Alma   (2020-10-26)


RFP included with Vibrio Compatible Plasmid Backbone


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 2049
    Illegal suffix found in sequence at 1
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 2049
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal NotI site found at 9
    Illegal NotI site found at 2055
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 2049
    Illegal XhoI site found at 1033
    Illegal XhoI site found at 1925
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 2049
    Illegal suffix found in sequence at 2
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 2049
    Illegal XbaI site found at 2064
    Illegal SpeI site found at 2
    Illegal PstI site found at 16
    Illegal AgeI site found at 2851
    Illegal AgeI site found at 2963
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

A C280T mutation was done on the origin of replication to allow compatibility with vibrio. The mutation allowed for the pSB1C3 backbone to resemble the pUC19 origin, which allows BioBricks to be expressed in vibrio.


Source

RFP BioBrick attached to the mutated backbone. Plasmic backbone contains a C280T mutation at the origin of replication to give it the same origin as pUC19 and therefore, make it compatible for replication with vibrio cells.

References