Difference between revisions of "Part:BBa K1321304"

Latest revision as of 14:35, 2 November 2014

pSEVA321-BB with J23117-RFP

This part contains Anderson promoter J23117 and RFP coding device in pSEVA321-BB backbone. pSEVA321-BB is a broad host range plasmid, capable of replication in E.coli and several other G- bacterial species. We have verified that it is also capable of replication in the cellulose producing bacterium Gluconacetobacter xylinus. RFP expression from BBa_K1321304 has been confirmed in E.coli and is currently being characterized in G.xylinus. BBa_K1321304 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 4476
Illegal SpeI site found at 37
Illegal PstI site found at 920
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 4476
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 4482
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 4476
23
INCOMPATIBLE WITH RFC[23]
Illegal prefix found in sequence at 4476
Illegal SpeI site found at 37
Illegal PstI site found at 920
25
INCOMPATIBLE WITH RFC[25]
Illegal prefix found in sequence at 4476
Illegal XbaI site found at 4491
Illegal SpeI site found at 37
Illegal PstI site found at 920
Illegal NgoMIV site found at 2095
Illegal NgoMIV site found at 2980
Illegal NgoMIV site found at 4091
Illegal NgoMIV site found at 4215
Illegal AgeI site found at 616
Illegal AgeI site found at 728
1000
COMPATIBLE WITH RFC[1000]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_K1321304 short</partinfo>
-Constitutive Anderson promoter J23102. Note the (Amp) plasmid backcone contains RFP reporter
+This part contains Anderson promoter J23117 and RFP coding device in pSEVA321-BB backbone. pSEVA321-BB is a broad host range plasmid, capable of replication in E.coli and several other G- bacterial species. We have verified that it is also capable of replication in the cellulose producing bacterium Gluconacetobacter xylinus. RFP expression from BBa_K1321304 has been confirmed in ''E.coli'' and is currently being characterized in ''G.xylinus''. BBa_K1321304 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.
 <!-- Add more about the biology of this part here
 ===Usage and Biology===
 <!-- -->