Difference between revisions of "Part:BBa K1321314"
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This is Anderson promoter '''J23105''' driving the expression of Elowitz RBS-mFRP in pSEVA331-Bb plasmid backbone (part BBa_K1321300 and is a member of the ''G.xylinus'' genetic engineering toolkit (parts BBa_K1321295 - BBa_K1321332). J23105 results in a low level of RFP expression in ''G.xylinus'', and is thus a weak promoter (see Figure 1). Further characterization of J23105 in ''G.xylinus'' is currently ongoing.  
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This is Anderson promoter '''J23105''' driving the expression of Elowitz RBS-mFRP in pSEVA331-Bb plasmid backbone (part BBa_K1321300 and is a member of the ''K. rhaeticus'' genetic engineering toolkit (parts BBa_K1321295 - BBa_K1321332) (see Figure 1).  
  
  
  
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[[File:IMG 4114 8b triple BBa J23105.JPG|600px|thumb|left|'''Figure 1.''' RFP expression from BBa_K1321314 in G.xylinus igem strain. Top three plates - transformed G.xylinus, bottom - untransformed control. Image taken under blue light box 5 days after transformation. G.xylinus was cultured on HS-agar plates, at 30degC inverted.]]
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[[File:IMG 4114 8b triple BBa J23105.JPG|600px|thumb|left|'''Figure 1.''' RFP expression from BBa_K1321314 in ''K. rhaeticus'' iGEM strain. Top three plates - transformed ''K. rhaeticus'', bottom - untransformed control. Image taken under blue light box 5 days after transformation. ''K. rhaeticus'' was cultured on HS-agar plates, at 30degC inverted.]]
  
  
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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.
  
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''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.
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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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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.
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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===

Revision as of 16:27, 6 May 2016

Anderson J23105-RFP in pSEVA331-Bb


This is Anderson promoter J23105 driving the expression of Elowitz RBS-mFRP in pSEVA331-Bb plasmid backbone (part BBa_K1321300 and is a member of the K. rhaeticus genetic engineering toolkit (parts BBa_K1321295 - BBa_K1321332) (see Figure 1).


Figure 1. RFP expression from BBa_K1321314 in K. rhaeticus iGEM strain. Top three plates - transformed K. rhaeticus, bottom - untransformed control. Image taken under blue light box 5 days after transformation. K. rhaeticus was cultured on HS-agar plates, at 30degC inverted.























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 3776
    Illegal SpeI site found at 37
    Illegal PstI site found at 920
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 3776
    Illegal NheI site found at 7
    Illegal NheI site found at 30
    Illegal SpeI site found at 37
    Illegal PstI site found at 920
    Illegal NotI site found at 913
    Illegal NotI site found at 3782
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 3776
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal prefix found in sequence at 3776
    Illegal SpeI site found at 37
    Illegal PstI site found at 920
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal prefix found in sequence at 3776
    Illegal XbaI site found at 3791
    Illegal SpeI site found at 37
    Illegal PstI site found at 920
    Illegal NgoMIV site found at 2095
    Illegal AgeI site found at 616
    Illegal AgeI site found at 728
  • 1000
    COMPATIBLE WITH RFC[1000]