''A. tumefaciens'' C58 TI-plasmid, partsregistry, Mr. Gene
This BioBrick is made to test our ''virA'' screening system and the strategy for directed evolution, respectively. It contains an unmutated ''virA'' under the control of a constitutive promoter and the screening device <partinfo>K389011</partinfo>.
medium strong constitutive promoters, only for testing the screening principle of mutated ''virA'' screening
This BioBrick contains a ''virA'' receptor from ''Agrobacterium tumefaciens'' C58 under the control of a constitutive promotor. We used it to mutate ''virA'' by error-prone PCR. For screening the mutants BBa_K389011 is also needed.
The ''virA'' gene is under the control a medium strong constitutive promotor.
synthetic ''virG'' gene, ''vir'' promotor from ''A. tumefaciens'' C58, partsregistry
This part contains: - a mutated ''virG'' response regulator from the virA/G receptor system from ''Agrobacterium tumefaciens'' which works in ''Escherichia coli'' without an ''rpoA'' gene from ''A. tumefaciens'' - a kanamycin resistance under the control of a ''vir'' promotor This device is used to screen ''virA'' receptor mutants. The ''virA'' receptor is mutated in the BBa_K389010 BioBrick. The better the receptor detects a substance, the more ''virG'' is phosphorylated and the stronger the kanamycin resistance from this BioBrick is expressed. The BioBricks BBa_K389010 and BBa_K389011 have to be used together in one cell.
medium strong constitutive promotor, terminator to keep basal expression of kanamycin resistance low
RBS (Elowitz 1999) -- defines RBS efficiency
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
Transcriptional terminator consisting of a 64 bp stem-loop.
''Agrobacterium tumefaciens'' C58, TI-plasmid
This is a gene for the ''virA'' receptor from ''Agrobacterium tumefaciens'' C58. An illegal ''PstI'' restriction site was removed by site-directed mutagenesis.
Illegal ''PstI'' restriction site removed
This is the ''virG'' response regulator from the virA/G receptor system. The virA/G receptor system is used by ''Agrobacterium tumefaciens'' to detect phenolic substances secreted by plants. In presence of these substances, ''virG'' is activated by the ''virA'' receptor and induces the transcription of genes under the control of a ''vir'' promotor. Normally, ''virG'' needs the ''rpoA'' RNA polymerase subunit from ''A. tumefaciens'' to work in ''Escherichia coli'' but this ''virG'' BioBrick is mutated, so works with the ''rpoA'' subunit from ''E. coli''.
synthesized by Mr. Gene, so optimated codon usage, removal of illegal restriction sites
This is a ''vir'' promotor from ''Agrobacterium tumefaciens'' C58. It is located on the TI-plasmid before the ''virB'' genes. It is activated by phosphorylated ''virG'' response regulator.
This is a kanamycin resistance part without promotor. It is made from the BioBrick BBa_P1003 and is used for screenings.
we removed the promotor from the kanamycin resistance gene in BioBrick BBa_P1003
Conversion of the iGEM parts registry to SBOL2.1