Difference between revisions of "Part:BBa K1321326"
 
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''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.
 
''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.
  
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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, with quality control provided (see Experience).  
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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.
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===

Latest revision as of 15:48, 6 May 2016

pTet01

This plasmid encodes for TetR generator and mRFP1 behind pTet promoter in pSEVA331Bb backbone, making mRFP1 inducible via tetracycline. See attached file for annotated sequence. This construct is a member of the Komagataeibacter genetic engineering toolkit (parts BBa_K1321295 - BBa_K1321332). File:IC14 pTet01.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 4685
    Illegal SpeI site found at 946
    Illegal PstI site found at 1829
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 4685
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 946
    Illegal PstI site found at 1829
    Illegal NotI site found at 1822
    Illegal NotI site found at 4691
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 4685
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 4685
    Illegal SpeI site found at 946
    Illegal PstI site found at 1829
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 4685
    Illegal XbaI site found at 4700
    Illegal SpeI site found at 946
    Illegal PstI site found at 1829
    Illegal NgoMIV site found at 3004
    Illegal AgeI site found at 1525
    Illegal AgeI site found at 1637
  • 1000
    COMPATIBLE WITH RFC[1000]