Inducible ampR expression combined with LasR, RhlI and aeBlue expression cassettes
The parts for this construct were derived from the 2013 distribution kit and the iGEM Team Uppsala.
This construct is the basis for a Quorum sensing system for two bacteria. The selection marker (amp-resistance) is induced by LasI(inducer)/LasR(activator) complex. The LasR is encoded on the construct itself, whereas the LasI is taken up from the media. Furthermore the RhlI synthetase is expressed to synthesize the autoinducer RhlI for the corresponding system in another cell (BBa_K1073035). By the activation of the amp-resistance the gene of interest (in this case the chromoprotein aeblue) is expressed. Together with BBa_K1073035 the construct forms a dependency between two cells (one with BBa_K10733034, the other with BBa_K1073035).
Inducible ampR expression combined with RhlR, LasI and eforRed expression cassettes
This construct is the basis for a Quorum sensing system for two bacteria. The selection marker (amp-resistance) is induced by a RhlI(inducer)/RhlR(activator) complex. The RhlR is encoded on the construct itself, whereas the RhlI is taken up from the media. Furthermore the LasI synthetase is expressed to synthesize the autoinducer LasI for the corresponding system in another cell (BBa_K1073034). By the activation of the amp-resistance the gene of interest (in this case the chromoprotein eforRed) is expressed. Together with BBa_K1073034 the construct forms a dependency between two cells (one with BBa_K10733034, the other with BBa_K1073035).
Double terminator consisting of BBa_B0010 and BBa_B0012
Inducible ampicillin resistance cassette + RhlR expression cassette
The biobricks for this part were taken from the 2013 distribution kit.
In complex with the inducer N-butyryl-HSL, the activator RhlR binds to the Rhl promoter and enables expression. This construct provides all the parts required to express the gene behind the rhl promoter. Only the inducer N-butyryl-HSL is missing, which usually is taken up from the media. By the regulation of the inducer's concentration in the media, the expression rate of the gene of interest (in this case ampicillin beta-lactamase) can be controlled.
Inducible ampicillin resistance cassette + LasR expression cassette
The parts for this construct were taken from the 2013 distribution kit.
In complex with the inducer PAI-1 (N-(3-oxododecanoyl)-homoserine lactone), the activator LasR binds to the Las promoter and enables expression. This construct provides all the parts required to express the gene behind the Las promoter. Only the inducer N-(3-oxododecanoyl)-homoserine lactone is missing, which usually is taken up from the media. By the regulation of the inducer's concentration in the media, the expression rate of the gene of interest (in this case ampicillin beta-lactamase) can be controlled.
Autoinducer synthase LasI + LVA with combined eforRed chromoprotein expression cassette
The biobricks for this part were derived from the 2013 distribution kit and the iGEM Team Uppsala.
This part is the basis for the expression of the Las auto inducer synthetase, which synthesizes LasI [N-(3-oxododecanoyl)-homoserine lactone]. LasI binds to the activator LasR. Together they activate the Las promoter. A constitutive expression of the chromoprotein eforRed is added to the construct in order to varify the expression of LasI synthetase. The construct has no promoter in front of the LasI synthetase gene. Therefore the user can choose the promoter himself (e.g. constitutive, induced, repressed, etc.)
Autoinducer synthase RhlI + LVA with combined aeBlue chromoprotein expression cassette
This part is the basis for the expression of the Rhl auto inducer synthetase, which synthesizes RhlI [N-(butyryl)-homoserine lactone]. RhlI binds to the activator RhlR. Together they activate the Rhl promoter. A constitutive expression of aeblue is added to the construct in order to varify the expression of RhlI synthetase. The construct has no promoter in front of the RhlI synthetase gene. Therefore the user can choose the promoter himself (e.g. constitutive, induced, repressed, etc.)
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.
Inducible ampicillin resistance cassette (regulated by RhlR + N-buturyl-HSL)
This part is a combination of BBa_K1073001 and BBa_K1073000, which themselves were constructed from biobricks of the 2013 distribution kit.
The promoter for this beta-lactonase is induced by N-(butyryl)-homoserine lactone. Therefore the expression of the ampicillin beta-lactonase is inducable by Rhl Inducer N-(butyryl)-homoserine lactone.
Inducible ampicillin resistance cassette (regulated by LasR + N-3-oxododecanoyl-HSL)
This part is a combination of BBa_K1073003 and BBa_K1073000, which themselves were constructed from parts of the 2013 distribution kit.
The promoter for this beta-lactonase is induced by N-(3-oxododecanoyl)-homoserine lactone. Therefore the expression of the ampicillin beta-lactonase is inducable by Las inducer N-(3-oxododecanoyl)-homoserine lactone.
Constitutively expressed transcription regulator RhlR + LVA
This part is a combination of the BBa_K1073006 and BBa_C0071.
The RhlR synthetase is constitutively expressed by a strong promoter.
Constitutively expressed transcription regulator LasR + LVA
This part is a combination of BBa_K1073006 and BBa_C0079, which are derived from Biobricks of the 2013 distribution kit.
The LasR activator is constitutively expressed by a strong promoter. It accepts the chemical signal AI-1 (made by lasI protein)
Autoinducer synthetase LasI ( producing N-3-oxododecanoyl-HSL) with RBS and double terminator
The part is a combination of BBa_B0032, BBa_C0078, and BBa_B0015, which were taken from the 2013 distribution kit.
To the lasI protein, which produces the chemical signal AI-1, a RBS and a double terminator was added to facilitate the construction of an expression system.
Autoinducer synthetase RhlI ( producing N-butyryl-HSL) with RBS and double terminator
This part is a combination of BBa_B0032, BBa_C0070 and BBa_B0015, which were taken from the 2013 distribution kit.
To the synthetase for the autoinducer N-butyryl-HSL, which binds to RhlR, (Part:BBa_C0071) a RBS and a double terminator was added to facilitate the construction of an expression system.
Constitutively expressed chromoprotein aeBlue
This part is a combination of BBa_K1073006 and BBa_K864401, which were taken from the 2013 distribution kit and the iGEM Team Uppsala.
The chromoprotein aeblue is constitutively expressed and can be used for the marking of cells for easier differentiation.
Constitutively expressed chromoprotein eforRed
This part is a combination of BBa_K1073006 and BBa_K592012, which were derived from the 2013 distribution kit and the iGEM Team Uppsala.
The chromoprotein eforRed is constitutively expressed and can be used for the marking of cells for easier differentiation.
RBS.3 (medium) -- derivative of BBa_0030
Weak1 RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-2" in figure 4-14 of thesis). Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
autoinducer synthetase for N-butyryl-HSL (BHL) and HHL
Autoinducer synthesis protein that produces N-butyryl-HSL which binds to RhlR, obtained from Pseudomonas aeruginosa.
Editing Check (1/28/04) (0) BB sites cleaned (1) ATG (2) TAATAA (3) Blast checks out (i.e., it's RhlI) (4) Codon changes OK (AA and tRNA usage) (5) Chassis might have conflict (w/ native autoinducer system?) (6) ssrA tag added (7) barcode added (8) Codon optimized for expression in enteric bacteria 2 silent point mutations have been included into the sequence at base 138 from A to G and at base 576 from G to C to remove internal EcoRI and PstI sites. Mutations were chosen to yield the same amino acid and codons of similar frequency in E. coli.
Debra Lin, Srini Devadas, David Gray, Ronny Krashinsky, and Chris Zheng Liu, Boopers
autoinducer synthetase for PAI from Pseudomonas aeruginosa
coding region for lasI protein, which produces the chemical signal AI-1
lasR activator from P. aeruginosa PAO1(+LVA)
Genbank: NC_002516 (www.ncbi.nlm.nih.gov)
coding region for lasR protein, which accepts chemical signal AI-1 (made by lasI protein)
Mutated 480 from g to a to remove PstI site
rhlR repressor/activator from P. aeruginosa PA3477 (+LVA)
Transcriptional regulator, in complex with N-butyryl-HSL, RhlR binds to the Rhl promoter
(1/28/04 Edit Jamboree) (0) No BB sites (1) ATG (2) TAATAA (3) Blast checks out (4) No codon changes (5) Maybe chassis compatability (check by experiment) (6) Codon usage not optimal (but looks OK) (7) ssrA tag added
The part was derived from BBa_P1002 ampicillin resistance cassette. The sequence for the beta-lactamase was gained by PCR of the original BBa_P1002 with appropiate primers.
This sequence codes for a beta-lactamase, which degrades ampicillin. There is no RBS or other information attached, only the sequence for the enzyme plus prefix and suffix.
The primer sequences can be found on iGEM Team Braunschweig's wiki.
Inducible promoter of the rhl quorum sensing system with RBS
Combined construct from BBa_R0071 and BBa_B0032, which were taken from the 2013 distribution kit.
This construct is derived from BBa_R0071 and B0032. It combines the rhl induced promoter with an RBS and can be used to clone it in front of a desired gene of interest, which shall be induced by rhl.
Inducible promoter of the las quorum sensing system with RBS
Thos part is a combination of BBa_K091117 and BBa_B0032, which were taken from the 2013 distribution kit.
This construct is derived from BBa_K091117 and BBa_B0032. It combines the las induced promoter with an RBS and can be used to clone it in front of a desired gene of interest, which shall be induced by las.
This part is a combination of BBa_J23106 and BBa_B0032, which were taken from 2013 distribution kit.
This part combines a constitutive promoter with an RBS and can be cloned in front of a gene of interest. It is a combination of BBa_J23106 and BBa_B0032.
Synthesized and codon optimized for E. coli by Bioneer Corp.
This chromoprotein eforRed naturally exhibits red color when expressed. The color is weaker than RFP, however. On agar plates and in liquid culture, the color is readily visible to naked eye in less than 24 hours of incubation. The DNA was codon-optimized for expression in E.coli and synthesized by the Korean company Bioneer Corporation.
Pseudomonas aeruginosa rhlAB promoter
Promoter activated by RhlR in concert with C4-HSL. C4-HSL is produced by RhlI, and acts as a quorum-sensing autoinducer. This is the natural sequence taken from Pseudomonas aeruginosa. Crosstalk: this promoter is also activated at a low level by LasR with its associated HSL.
The -10 and -35 sites are labeled as identified in [Pearson97]. The part begins with the RhlR binding site, and (rather arbitrarily) ends with the first transcribed base (+1).
The sequence of the LasI upstream regulatory region is available at http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=176640
This pLas' promoter contains the regulatory region upstream of the LasI gene in Pseudomonas aeruginosa. K091117 includes a LasR binding site as well as a -10 region modified from CATAAA to TATAAA to reflect our pLasXOR promoter design. Activation is expected to occur in the presence of the autoinducer PAI-1 and the LasR protein. The pLas' promoter serves as a control for the evaluation of the pLasXOR promoter, part BBa_K091118.
To minimize leaky transcription, a secondary transcriptional start site was excluded from the design.
Conversion of the iGEM parts registry to SBOL2.1