This expression casette constitutively produces the LuxR regulatory protein that can respond to the quorum sensing molecule 3OC6. It also produces a LacI repressor with a mutation at position 12 that results in an isoleucine. The mutant repressor has been shown to have an increased affinity for its operator.
This is a composite part of pBAD, B0034, and LuxR. LuxR are is the receiver protein for the quorum sensing molecule AI-1 produce by LuxI.
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.
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.
Transcriptional terminator consisting of a 64 bp stem-loop.
The basic sequence comes from Ecogene.com
This is a variation of the LacI protein that acts as a repressor for the pLac promoter and is repressed by IPTG. It has a point mutation at the third codon where proline is replaced by tyrosine. This missense mutation causes an increased binding affinity of the LacI_I12 protein with the pLac promoter.
Because there are many codons that code for the amino acid tyrosine, we chose to use the TAT codon, because it was the most common tyrosine codon found throughout the rest of the LacI wild-type gene sequence.
V. fischeri <genbank>AF170104</genbank>
In complex with HSL, LuxR binds to the Lux promoter, activating transcription from Pr <bb_part>BBa_R0062</bb_part>, and repressing transcription from Pl <bb_part>BBa_R0063</bb_part>. The lux cassette of V. fischeri contains a left and a right promoter. The right promoter gives weak constitutive expression of downstream genes.This expression is up-regulated by the action of the Lux activator, LuxR complexed to HSL. Two molecules of LuxR protein form a complex with two molecules the signalling compound homoserine lactone (HSL). This complex binds to a palindromic site on the promoter, increasing the rate of transcription.
2 silent point mutants were introduced in the coding sequence to remove internal XbaI and PstI sites. Mutation sites were chosen to replace codons commonly used in E. coli with codons used at a similar frequency.
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr
Conversion of the iGEM parts registry to SBOL2.1