pLacI Regulated Lumazine Synthase with Transciptional Terminator and pBAD
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K542001 BBa_K542001] and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K542003 BBa_K542003] were both assembled by the Lethbridge 2011 team.
Made by assembling [http://partsregistry.org/wiki/index.php?title=Part:BBa_K542001 BBa_K542001] with [http://partsregistry.org/wiki/index.php?title=Part:BBa_K542003 BBa_K542003]. Note that the pLacI promoter ([http://partsregistry.org/wiki/index.php?title=Part:BBa_R0010 BBa_R0010]) is regulating the Lumazine Synthase gene, not the pBAD promoter ([http://partsregistry.org/Part:BBa_I13453 BBa_I13453]) which is downstream. Therefore, Lumazine Synthase production is IPTG inducible. The microcompartment portion to the "Co-localization Construct".
The microcompartment portion to the "Co-localization Construct".
[http://partsregistry.org/Part:BBa_J04500 BBa_J04500] obtained from iGEM distribution plate and [http://partsregistry.org/wiki/index.php?title=Part:BBa_K542000 BBa_K542000] was assembled by Lethbridge 2011 team.
Made by assembling [http://partsregistry.org/Part:BBa_J04500 BBa_J04500] with [http://partsregistry.org/wiki/index.php?title=Part:BBa_K542000 BBa_K542000] IPTG inducible Lumazine Synthase. Intermediate part to assemble the "Co-localization Construct".
Intermediate part to assemble the "Co-localization Construct".
[http://partsregistry.org/wiki/index.php?title=Part:BBa_I13453 BBa_I13453] and [http://partsregistry.org/Part:BBa_P0440 BBa_P0440] were both obtained from iGEM distribution plates.
Made by assembling [http://partsregistry.org/wiki/index.php?title=Part:BBa_I13453 BBa_I13453] with [http://partsregistry.org/Part:BBa_P0440 BBa_P0440]. Production of TetR is regulated by the pBAD promoter([http://partsregistry.org/Part:BBa_I13453 BBa_I13453]). TetR is an inhibitor of the constitutively "on" promoter pTet--[http://partsregistry.org/wiki/index.php?title=Part:BBa_R0040 BBa_R0040]). Therefore, this part may be used in conjunction with the pTet promoter as an "inverter"; pTet will be "turned off" in the presence of arabinose. Arabinose -> pBAD "on" -> TetR -> pTet "off" Intermediate part to assemble the "Co-localization Construct".
Lumazine Synthase with Transciptional Terminator (No Promoter)
[http://partsregistry.org/wiki/index.php?title=Part:BBa_K249002 BBa_K249002] and [http://partsregistry.org/Part:BBa_B0015 BBa_B0015] were obtained from iGEM distribution plates.
Composite part: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K249002 BBa_K249002] + [http://partsregistry.org/wiki/index.php/Part:BBa_B0010 BBa_B0010] + [http://partsregistry.org/wiki/index.php/Part:BBa_B0010 BBa_B0010] + [http://partsregistry.org/wiki/index.php/Part:BBa_B0012 BBa_B0012] Made by assembling [http://partsregistry.org/wiki/index.php?title=Part:BBa_K249002 BBa_K249002] with [http://partsregistry.org/Part:BBa_B0015 BBa_B0015] Since this part is lacking the promoter, Lumazine Synthase production may be regulated by the addition different promoters upstream. Regulation of Lumazine Synthase will be dependent on the promoter being utilized.
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.
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.
Transcriptional terminator consisting of a 64 bp stem-loop.
Double terminator consisting of BBa_B0010 and BBa_B0012
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.
Lumazine Synthase is an enzyme which creates Lumazine, a product which aggregates forming a hollow spheroid which can act as a mirocompartment, or artificial organelle. The Lumazine forms negatively charged pores, which can be used to introduce proteins. The proteins which are being introduced into the microcompartment must be equipped with an Arginine Tag.
point mutations of EcoRI and PstI sites?
Conversion of the iGEM parts registry to SBOL2.1