RhlR, PrhlR + LacI modified , PLac + TetR, pTetR + RFP and LuxI
Source of LacIm : Basu et al."A synthetic multicelluar system for programmed pattern formation", Nature. 2005 Apr 28;434(7037):1130-4
Sequence containing : - RhlR controlled by a constitutive promoter - LacI modified with a RhlR promoter - TetR controlled by a Lac Promoter - RFP controlled by a TetR promoter - LuxI is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the TetR promoter. The transcription of RFP and LuxI is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
The Plac insert was PCR'd from the MG1655 strain of E.coli K12.
Inverting regulatory region controlled by LacI (<bb_part>BBa_C0010</bb_part>, <bb_part>BBa_C0011</bb_part>, etc.) The pLac regulatory region is a 243 base-pair sequence with standard BioBrick prefix and suffix sections on its ends. It contains two protein binding sites: CAP, which is generally present in E.coli and is assocciated with cell health and availability of glucose., and LacI, the Lac inhibitor <bb_part>BBa_C0010</bb_part> which binds in an dimerized cooperative manner to inhibit the transcription of the protein that follows. In the presence of lactose or IPTG, an analog of lactose, LacI is unable to correctly bind and inhibit transcription. This allows <bb_part>BBa_R0010</bb_part> to be used as a inverter or as a detector of lactose or IPTG.
LacI binds to this regulator. This part is incompatible with species containing active LacI coding regions. Lactose and IPTG disable the operation of LacI and this regulator. This part is incompatible with environments containing lactose or lactose analogs.
Double terminator consisting of BBa_B0010 and BBa_B0012
TetR cds with RBS and Terminator + pTetR, RFP with RBS + LuxI with RBS and terminator
Combination of the following parts : BBa_K092000, BBa_K092300 and BBa_K092400: The TetR sequence (with a ribosome binding site and terminator)[BBa_K09200] is bound to the part containing the promoter of TetR and the red fluorescent protein (RFP) with a ribosome binding site. A luxI part (with RBS and terminator) is added at the end of the cds of the RFP. As no terminator was placed after the RFP, they both depends on the TetR promoter. The transcription of RFP and LuxI is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
RhlR controlled by a constitutive promoter, and LacI modified with a Rhl R promoter
The LacIm part comes from part BBa_K092800, from Basu et al."A synthetic multicelluar system for programmed pattern formation", Nature. 2005 Apr 28;434(7037):1130-4
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.
rhlR repressor/activator from P. aeruginosa PA3477 (no LVA)
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).
Constitutive Promoter, Medium Transcription
Vikram Vijayan, Allen Hsu, Lawrence Fomundam
LuxI with RBS and Terminator build by 2-steps PCR.
TetR cds with RBS and Terminator + pTetR, RFP with RBS
Composite part : combination of the part BBa_K092000 and the BBa_K092300 : The TetR sequence (with a ribosome binding site and terminator) is bound to the part containing the promoter of TetR, a ribosome binding site and the red fluorescent protein (RFP). Thus, the production of RFP directly depends on the production of TetR. The transcription of RFP is inhibited when TetR is present. The production of TetR depends on the promoter you put in front of it.
Coding sequence for LacI modified with a different rate of translation than LacI, RBS
LacIm from Basu et al."A synthetic multicelluar system for programmed pattern formation", Nature. 2005 Apr 28;434(7037):1130-4
TetR coding sequence with RBS and terminator
Coding sequence of the Tetracycline repressor with the ribosome binding site and a double terminator. The TetR comes from transposon Tn10 and has a LVA tail (SsrA) [cf. C0040 part].
Coding sequence of the RFP with a ribosome binding site and pTetR as promoter. Thus, the transcription of RFP is inhibited when TetR is present.
**highly** engineered mutant of red fluorescent protein from Discosoma striata (coral)
Campbell et al., PNAS v99 p7877 URL
monomeric RFP: Red Fluorescent Protein. Excitation peak: 584 nm Emission peak: 607 nm
TAATAA double stop codon added (DE). Four silent mutations made to remove three EcoRI sites and one PstI site: A28G, A76G, A349G, G337A.
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
tetracycline repressor from transposon Tn10 (+LVA)
Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999)
Coding region for the TetR protein without the Ribosome Binding Site. Modified with an LVA tail for rapid degradation of the protein and faster fall time for the emission. TetR binds to the pTet regulator (BBa_R0040). aTc (anhydrotetracycline) binds to TetR and inhibits its operation.
References (unparsed) here: Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999). Lutz R, Bujard H., Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 1997 Mar 15;25(6):1203-10. PMID: 9092630 References (unparsed) here: Elowitz, M. B. Transport, Assembly, and Dynamics in Systems of Interacting Proteins. Thesis, Princeton Univ., Princeton (1999). Lutz R, Bujard H., Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res. 1997 Mar 15;25(6):1203-10. PMID: 9092630 BBa_C0040 TetR Protein is based on the TetR sequence from Elowitz's repressilator. It has been modified to include a rapid degradation LVA tail, and includes the BioBrick standard assembly head and tail restriction sites. The RBS has been removed. The stop codon has been changed from TAA to a double stop codon TAATAA.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman.
Conversion of the iGEM parts registry to SBOL2.1