Composite

Part:BBa_K1321324

Designed by: Michael Florea   Group: iGEM14_Imperial   (2014-10-08)

pLux01

This plasmid encodes for LuxR generator and mRFP1 behind pLux promoter, making mRFP1 inducible via AHL, in pSEVA331Bb backbone. See attached file for annotated sequence. It is a member of the Komagataeibacter genetic engineering toolkit (parts BBa_K1321295 - BBa_K1321332). File:IC14 pLux01.gb



Komagataeibacter toolkit was designed to ease genetic engineering of cellulose-based biomaterials using the cellulose synthesizing bacterium Komagataeibacter rhaeticus iGEM (parts Bba_K1321305 and BBa_K1321306). As no registry parts had been tested in K. rhaeticus, the aim of this toolkit was to determine the parts usable in K. rhaeticus and to characterize them in this host. pSEVA331-Bb is a non-standard broad host range plasmid capable of replication in K. rhaeticus and E. coli (a shuttle vector) and was selected because the registry's standard plasmid backbone pSB1C3 can not be used for K. rhaeticus engineering.

NOTE: Because the registry's standard plasmid backbone pSB1C3 is not capable of replication in Komagataeibacter or Gluconacetobacter species, the Komagataeibacter genetic engineering toolkit is housed mainly in pSEVA331-Bb. pSEVA331-Bb is a non-standard backbone, which therefore can't be quality controlled by and maintained in the Registry. However, in order to make the Komagataeibacter toolkit available for the synthetic biology community, Imperial iGEM 2014 team has made it freely available in AddGene or upon request.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4785
    Illegal SpeI site found at 1046
    Illegal PstI site found at 1929
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4785
    Illegal SpeI site found at 1046
    Illegal PstI site found at 1929
    Illegal NotI site found at 1922
    Illegal NotI site found at 4791
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4785
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4785
    Illegal SpeI site found at 1046
    Illegal PstI site found at 1929
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4785
    Illegal XbaI site found at 4800
    Illegal SpeI site found at 1046
    Illegal PstI site found at 1929
    Illegal NgoMIV site found at 3104
    Illegal AgeI site found at 1625
    Illegal AgeI site found at 1737
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 987


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