Inducible ampicillin resistance cassette + LuxR expression cassette
The biobricks for this construct were taken from the 2013 distribution kit.
In complex with the inducer 3-oxo-hexanoyl-HSL, the activator LuxR binds to the Lux promoter and enables expression. This construct provides all the parts required to express the gene behind the Lux promoter. Only the inducer 3-oxo-hexanoyl-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.
Double terminator consisting of BBa_B0010 and BBa_B0012
Inducible ampicillin resistance cassette (regulated by LuxR + N-3-oxohexanyol-HSL)
This part is a combination of BBa_K1073002 and BBa_K1073000, which themselves were constructed from biobricks of the 2013 distribution kit.
The promoter for this beta-lactonase is induced by N-(3-oxohexanoyl)-homoserine lactone. Therefore the expression of the ampicillin beta-lactonase is inducable by Lux inducer N-(3-oxohexanoyl)-homoserine lactone.
Constitutively expressed transcription regulator LuxR + LVA
This part is a combination of BBa_K1073006 and BBa_C0062, which are derived from Biobricks of the 2013 distribution kit.
The LuxR synthetase is constitutively expressed by a strong promoter.
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.
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
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 lux quorum sensing system with RBS
The construct was combined from BBa_R0062 and BBa_B0032, which were taken from the 2013 distribution kit.
This construct is derived from BBa_R0062 and BBa_B0032. It combines the lux 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 lux.
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.
Promoter (luxR & HSL regulated -- lux pR)
Promoter activated by LuxR in concert with HSL 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 LuxR activator protein complexed with the autoinducer, 3-oxo-hexanoyl-HSL. Two molecules of LuxR protein form a complex with two molecules of the signalling compound homoserine lactone (HSL). This complex binds to a palindromic site on the promoter, increasing the rate of transcription.
This promoter is based on the Vibrio fischeri quorum sensing gene promoters. Two genes LuxI and LuxR and transcribed in opposite directions as shown below. The original sequence from which the parts <bb_part>BBa_R0062</bb_part> and <bb_part>BBa_R0063</bb_part> were derived is shown in the picture below. <img src="<bb_file>Image1.gif</bb_file>" width="614" height="362">
Conversion of the iGEM parts registry to SBOL2.1