pLux-Lac:GFP:Term:Term:PluxR/cI-OR :mCherry:Term:PtetR:LuxR:Term:Fixed-pLux/cI:LuxI
LuxI expression controlled by the hybrid promoter Part:BBa_K415032 which is a tighter version of the promoter Part:BBa_R0065.
LuxI expression controlled by a hybrid promoter that is activated by a LuxR-AHL complex and inhibited by cI. The "fixed" promoter is Part:BBa_K415031 which is a more tightly controlled (less noisy) version of BBa_R0065.
Pluxlac RBS-GFP TT PluxCI RBS-mCherry TT PtetR RBS-LuxR TT
This part displays red and green fluorescence under specific conditions. Fluorescence is activated by presence of both LuxR and 3OC6HSL. The LuxR is normally constitutively expressed, but can be repressed by TetR and derepressed by aTc. Green fluorescence is repressed by LacI and red fluorescence is repressed by CI. This part is intended to be used along with the LacI/CI bistable toggle switch by Jim Collins.
pLux/cI-OR : RBS-mCherry : Term : p(tetR) : RBS-luxR : Term
This composite is composed of the parts BBa_K415000 and BBa_KS03119. It can produce both luxR and mCherry. The production of luxR is repressed by aTc. The production of mCherry is activated by c6-AHL and luxR, and repressed by cI. This is a test part that generates a reporter.
The luxR generator is put second, in case the terminator of BBa_K415000 is leaky.
This is a hybrid promoter synthesized from a portion of the Vibrio Fischerii Lux operon and the cI/cro switch of Lambda bacteriophage.
This part is a promoter that was designed as a correction to the anomalous BBa_R0065 BioBrick. This part consists of a Lux Box binding site for the LuxR-C6HSL Complex; in addition, it contains two operator sites for the cI protein appropriated from the Lambda bacteriophage. Because basal expression without the LuxR-C6HSL complex present is expected to be minimal, and because the cI operator sites are positioned so as to block the critical -10 transcription initiation site on the promoter, if cI is high, the promoter is off; if AHL is low and cI is absent, the promoter is leaky; if AHL is high and cI is absent, the promoter is on.
This promoter was designed entirely as a result of issues with the R0065 promoter, which was essentially designed as a literal fusion of two promoters that happened to contain twin -10 transcription initiation sites. This resulted in a promoter that was not leaky without the presence of AHL, but constitutively on. This newly designed promoter should properly follow the logic detailed above.
pLux-Lac Hybrid with GFP Reporter and Double Terminator
Ligation of BBa_I751502 upstream and BBa_E0840 downstream.
Hybrid Promoter with two LacO sites and pLux-Lac sites for Quorum Sensing reporting via GFP. Lux promoter is foremost followed by first operator, then pLac and second LacO site. Comprised of parts BBa_I751502 upstream and BBa_E0840 downstream.
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.
V. fischeri <genbank>AF170104</genbank>
Synthesizes 3OC6HSL, which binds to LuxR. 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 repressor, LuxR. Two molecules of LuxR protein form a complex with two molecules the signalling compound HSL. This complex binds to a palindromic site on the promoter, increasing the rate of transcription.
An LVA tail (sequence: AANDENYALVA) was added to increase protein degradation. .
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr
Transcriptional terminator consisting of a 64 bp stem-loop.
RBS.1 (strong) -- modified from R. Weiss
Strong RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("orig" in figure 4-14 of Ron Weiss thesis). 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 Brian Chow
green fluorescent protein derived from jellyfish Aequeora victoria wild-type GFP (SwissProt: P42212
GFP (mut3b) [note that this part does not have a barcode]
sense TWO INPUTS, activation by AHL and repression by LacI.
Compared with existing BioBrick hybrid promoter R0065, this promoter is different in that its sequence is much shorter with lux box and -35 region overlapping, and that it is repressed by LacI, not cI. Also, the output is clearly detected as shown in Fig. 3.
http://partsregistry.org/wiki/index.php?title=Part:BBa_R0065 http://partsregistry.org/wiki/index.php?title=Part:BBa_J06702
This is a BioBrick composed of parts BBa_R0065 and BBa_J06702. This composite produces the mCherry fluorescent protein. It is activated by the presence of LuxR and AHL, and it is repressed by cI.
Lutz, R., Bujard, H., Nucleic Acids Research (1997) 25, 1203-1210.
Sequence for pTet inverting regulator driven by the TetR protein.
BBa_R0040 TetR-Regulated Promoter is based on a cI promoter. It has been modified to include two TetR binding sites and the BioBrick standard assembly head and tail restriction sites.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman
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.
mCherry from Roger Y. Tsien's lab, altered to be BioBrick compatible
mRFP (DsRed) derived, altered to be a biobrick by removing a PstI site and adding in the ends.
Made a point mutation to eliminate a PstI site in the middle and then added BioBrick ends using PCR. Sequenced using primers that bind to the pSB1A2 plasmid on either side of the brick.
Promoter (lambda cI and luxR regulated -- hybrid)
cI repressor negatively regulates this promoter and LuxR activates its transcription.The effect of cI is dominant over LuxR. This part is based on the LuxR and cI repressor regulated hybrid promoter designed and tested by Ron Weiss. It requires the binding of two cI repressor dimers for maximal repression and contains two cI repressor binding sites namely, OR1 and OR2. This promoter is leaky in the sense that 'some' transcription is seen in the absence of both cI and LuxR. LuxI cI activity of promoter + + zero + - maximum - + zero - - leaky (no quantitative information)
This part was designed based on the LuxR and cI repressor regulated hybrid promoter tested by Ron Weiss and the LuxR-LuxICDABE sequence annotated by Tom Knight <genbank>AF170104</genbank>.
Conversion of the iGEM parts registry to SBOL2.1