Difference between revisions of "Part:BBa K1321328"

Revision as of 16:11, 6 May 2016

J-sRNA-331Bb

This construct expresses an sRNA and E. coli Hfq behind an AHL-inducible promoter, which leads to suppression of cellulose production in Komagataeibacter rhaeticus iGEM. The 5' of the sRNA is complementary to K. rhaeticus iGEM UGPase mRNA RBS region, and 3' has an E. coli Hfq binding region - upon expression, it complexes with UGPase mRNA, and leads to Hfq-mediated translational repression. This construct is a member of the Komagataeibacter 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 4541
Illegal EcoRI site found at 1297
Illegal SpeI site found at 1207
Illegal PstI site found at 1685
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 1297
Illegal EcoRI site found at 4541
Illegal SpeI site found at 1207
Illegal PstI site found at 1685
Illegal NotI site found at 1678
Illegal NotI site found at 4547
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 1297
Illegal EcoRI site found at 4541
Illegal XhoI site found at 1145
Illegal XhoI site found at 1151
23
INCOMPATIBLE WITH RFC[23]
Illegal prefix found in sequence at 4541
Illegal EcoRI site found at 1297
Illegal SpeI site found at 1207
Illegal PstI site found at 1685
25
INCOMPATIBLE WITH RFC[25]
Illegal prefix found in sequence at 4541
Illegal EcoRI site found at 1297
Illegal XbaI site found at 4556
Illegal SpeI site found at 1207
Illegal PstI site found at 1685
Illegal NgoMIV site found at 2860
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 1198
Illegal BsaI.rc site found at 987

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 This construct expresses an sRNA and <i>E. coli</i> Hfq behind an AHL-inducible promoter, which leads to suppression of cellulose production in <i>Komagataeibacter rhaeticus</i> iGEM. The 5' of the sRNA is complementary to <i>K. rhaeticus</i> iGEM UGPase mRNA RBS region, and 3' has an <i>E. coli</i> Hfq binding region - upon expression, it complexes with UGPase mRNA, and leads to Hfq-mediated translational repression.
-This construct is a member of the ''G.xylinus'' genetic engineering toolkit (parts BBa_K1321295 - BBa_K1321332).
+This construct is a member of the ''Komagataeibacter'' 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.
+''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 ''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.
+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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 ===Usage and Biology===