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.
Double terminator consisting of BBa_B0010 and BBa_B0012
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 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)
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.
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
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 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.
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.
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