Composite

Part:BBa_K1321329

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

GFP-LVA under TetR control in pSEVA331-Bb


This device consists of a TetR generator and a GFP with LVA tag under control of a TetR promoter in pSEVA331-Bb plasmid backbone (part BBa_K145280). This construct is a member of the G.xylinus genetic engineering toolkit (parts BBa_K1321295 - BBa_K1321332).



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

NOTE: Because the registry's standard plasmid backbone pSB1C3 is not capable of replication in Gluconacetobacter species, the G.xylinus 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 G.xylinus toolkit available for the synthetic biology community, Imperial iGEM 2014 team has made it freely available upon request, with quality control provided (see Experience). To request, please contact Imperial iGEM 2014 team.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal prefix found in sequence at 4748
    Illegal suffix found in sequence at 1877
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4748
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 1878
    Illegal PstI site found at 1892
    Illegal NotI site found at 1885
    Illegal NotI site found at 4754
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4748
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4748
    Illegal suffix found in sequence at 1878
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4748
    Illegal XbaI site found at 4763
    Illegal SpeI site found at 1878
    Illegal PstI site found at 1892
    Illegal NgoMIV site found at 3067
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 1616


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