Difference between revisions of "Part:BBa K1321318"

Latest revision as of 16:40, 6 May 2016

Anderson J23114-RFP in pSEVA331-Bb

This is Anderson promoter J23114 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).

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]

@@ Line 2: / Line 2: @@
 <partinfo>BBa_K1321318 short</partinfo>
-This is Anderson promoter '''J23114''' 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). RFP expression from BBa_K1321318 has been confirmed in ''E.coli''. BBa_K1321332 is currently being characterized in ''G.xylinus''.
+This is Anderson promoter '''J23114''' 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).
+''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.
-''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.
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