vir promoter + sensitivity tuner + luciferase
* luciferase from pGL4.10luc2 (Promega) * sensitivity tuner part, terminators and constitutive promoter from the partsregistry * ''virG'' was synthesized by Mr. Gene * ''vir'' promoter from ''Agrobacterium tumefaciens'' C58, TI-plasmid
This part combines a mutated ''virG'' response regulator from the VirA/G receptor system under the control of a constitutive promoter with a sensitivity tuner before a luciferase gene under the control of a ''vir'' promoter. The sensitivity tuner enhances a signal from a promoter so it enhances the readout luciferase.
sensitivity tuner to enhance the luciferase signal
* luciferase from pGL4.10luc2 (Promega) * the rest from the partsregistry
This is a modified <partinfo>I746370</partinfo> sensitivity tuner. Instead of a GFP reporter, this one has a firefly luciferase (from Promega's pGL4.10luc2 vector) as one reporter gene. The arabinose promoter has been removed from the original part so now it is possible to assemble this part behind any promoter to get an enhanced luciferase signal.
arabinose promoter removed, GFP exchanged with luciferase compared to the original part <partinfo>I746370</partinfo>
Double terminator with two copies of <bb_part>BBa_B0010</bb_part>
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.
plasmid DNA supplied by Prof. Richard Calendar, University of California.
The ogr activator taken from P2 phage acts on a class of inducible promoters (parts I746360 to I746365), inducing their activity to varying degrees. The part sequence does already contain a ribosome binding site (B0034)!
The part does contain a RBS (B0034) already.
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.
**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)
Campbell et al., PNAS v99 p7877 URL
monomeric RFP: Red Fluorescent Protein. Excitation peak: 584 nm Emission peak: 607 nm
TAATAA double stop codon added (DE). Four silent mutations made to remove three EcoRI sites and one PstI site: A28G, A76G, A349G, G337A.
Double terminator consisting of BBa_B0010 and BBa_B0012
The original source is the P2 phage genome. For our work the DNA came from plasmids, kindly supplied to us by Prof. Calendar, University of California (Berkeley): Bacteriophage PSP3 and fR73 Activator Proteins: Analysis of Promoter Specificities; Julien and Calendar, 1996; JOURNAL OF BACTERIOLOGY, Oct. 1996, p. 5668???5675
This is the PF promoter taken from the P2 phage genome. It is an inducible promoter that is activated by a class of activators, including P2 ogr (I746350), PSP3 pag (I746351) and phiR73 delta (I746352). These different activators should cause different levels of activity of the PF promoter.
no special considerations seemed necessary
A luciferase from Promega's pGL4.10luc2 vector.
sequence of this vector is already published by Promega
Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
Transcription terminator for the E.coli RNA polymerase.
Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG). Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
Conversion of the iGEM parts registry to SBOL2.1