Sources: RBS: BBa_B0034 p8-leader sequence: First 23 amino acids of p8, exact sequenced derived from M13KE. GR1 coil: from Wang C, Zhong P, Wang X. Adapter-directed display systems. In Vitro. 2007;17(12). linker sequence: Linker 18 from Weiss et al. Mutational analysis of the major coat protein of M13 identifies residues that control protein display. 2000 opti-p8:from Weiss et al. 2000, derived from M13KE The source of the p8-leader sequence is M13KE. The optimized p8 is derived from M13KE3 (see Weiss et al. Mutational analysis of the major coat protein of M13 identifies residues that control protein display. 2000).
This part is a composite part of BBa_K415010 (PluxR/cI : RBS : mCherry : Term : PtetR : RBS : LuxR:Term : Plux/cI) with a p8-GR1 fusion appended, where p8 is the major coat protein of M13 bacteriophage and GR1 is a leucine zipper coil that can bind to corresponding zipper coil, GR2. This part is designed to produce the fusion, which can then be displayed on the phage coat when used in a helper phage system. The p8-GR1 part consists of an RBS, the p8 leader sequence, the zipper coil, an HA tag, a linker sequence, followed by a phage display optimized p8.
Linker 18 picked as one of the 15 amino acid linkers to enhance high copy protein display. opti-p8 was chosen for the same reason.
This is part K415006 with an appended Plux/cI promoter (R0065)
This composite is composed of the parts BBa_K415000 and BBa_S03119. It can produce both luxR and mCherry. The production of luxR is normally constitutive, but can be repressed by TetR and derepressed by aTc. The production of mCherry is activated by c6-AHL and luxR, and repressed by cI. This is a test part that generates a reporter. It also contains an additional R0065 hybrid promoter appended after the LuxR : Term : Term.
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.
Promoter (lambda cI and luxR regulated -- hybrid)
cI repressor negatively regulates this promoter and LuxR activates its transcription.The effect of cI is dominant over LuxR. This part is based on the LuxR and cI repressor regulated hybrid promoter designed and tested by Ron Weiss. It requires the binding of two cI repressor dimers for maximal repression and contains two cI repressor binding sites namely, OR1 and OR2. This promoter is leaky in the sense that 'some' transcription is seen in the absence of both cI and LuxR. LuxI cI activity of promoter + + zero + - maximum - + zero - - leaky (no quantitative information)
This part was designed based on the LuxR and cI repressor regulated hybrid promoter tested by Ron Weiss and the LuxR-LuxICDABE sequence annotated by Tom Knight <genbank>AF170104</genbank>.
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr
pLux/cI-OR : RBS-mCherry : Term : p(tetR) : RBS-luxR : Term
This composite is composed of the parts BBa_K415000 and BBa_KS03119. It can produce both luxR and mCherry. The production of luxR is repressed by aTc. The production of mCherry is activated by c6-AHL and luxR, and repressed by cI. This is a test part that generates a reporter.
The luxR generator is put second, in case the terminator of BBa_K415000 is leaky.
Fusion of M13 p8 and GR1 leucine zipper. Includes leader sequence and HA tag.
http://partsregistry.org/wiki/index.php?title=Part:BBa_R0065 http://partsregistry.org/wiki/index.php?title=Part:BBa_J06702
This is a BioBrick composed of parts BBa_R0065 and BBa_J06702. This composite produces the mCherry fluorescent protein. It is activated by the presence of LuxR and AHL, and it is repressed by cI.
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.
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
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.
mCherry from Roger Y. Tsien's lab, altered to be BioBrick compatible
mRFP (DsRed) derived, altered to be a biobrick by removing a PstI site and adding in the ends.
Made a point mutation to eliminate a PstI site in the middle and then added BioBrick ends using PCR. Sequenced using primers that bind to the pSB1A2 plasmid on either side of the brick.
Conversion of the iGEM parts registry to SBOL2.1