Cell density controller: pCon 0.15->luxI-LVA+pCon 0.70->luxR+pLux/Tet->ccdB-LVA
Cell Death = (Cell Density * pCon 0.15) + Input AHL
Input: AHL Output1: AHL Output2: Cell Density Design: pCon 0.15(K176008) generates LuxI-LVA, LuxI-LVA generates Output1 AHL, the concentration of AHL in the medium depends on both LuxI-LVA level per cell and cell density. pCon 0.70(J23101) generates LuxR. pLux/Tet(K176000) generates ccdB-LVA to kill the cell, activated by LuxR and (Output1 AHL generated by LuxI-LVA + Input AHL), repressed by TetR and no aTc.
cell density->PoPS: pCon 0.15->luxI-LVA+pCon 0.70->luxR+pLux/Tet->PoPS
Input: AHL Output1: AHL Output2: PoPS Design: pCon 0.15(K176008) generates LuxI-LVA, LuxI-LVA generates Output1 AHL, the concentration of AHL in the medium depends on both LuxI-LVA level per cell and cell density. pCon 0.70(J23101) generates LuxR. pLux/Tet(K176000) generates Output2 PoPS, activated by LuxR and (Output1 AHL generated by LuxI-LVA + Input AHL), repressed by TetR and no aTc.
strong RBS + ccdB-LVA coding + strong terminator
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.
TetR repressible AHL->PoPS Receiver (Very High LuxR): pCon 0.70->luxR+pLux/Tet->PoPS
AHL -> PoPS Receiver TetR repressible Similar to F2620, but use pLux/Tet hybrid promoter.
Input: AHL Output: PoPS Design: pCon 0.70(J23101) generate LuxR pLux/Tet(K176000) generate PoPS output, activated by LuxR and AHL, repressed by TetR and no aTc
PCR from K145151. Forward Primer: VF2(G00100) Reverse Primer: GTTT ACTAGT A TTATTA AGC TAC TAA AGC GTA GTT TTC GTC GTT TGC AGC tattccccagaacatcaggtta Digest with EcoRI and SpeI, ligate into pSB1A3.
Add LVA degradation tag (widely used in C0040, C0051, C0061...) to ccdB(K145151) C-terminal: [K145151 without stop codon] GCT GCA AAC GAC GAA AAC TAC GCT TTA GTA GCT [TAA TAA] A A N D E N Y A L V A
constitutive promoter family member J23115 actual sequence (pCon 0.15)
The sequence of J23115 is wrong according to sequencing result of the registry and ours. This part K176008 is created for the actual promoter. http://partsregistry.org/cgi/sequencing/one_blast.cgi?id=3647 http://partsregistry.org/cgi/sequencing/one_blast.cgi?id=1209 J23115: tttatagctagctcagCcctTggtacaatgctagc K176008: tttatagctagctcagTcctAggtacaatgctagc
pLux/Tet Hybrid Promoter: (LuxR+,TetR-)->PoPS
sense TWO INPUTS, activation by AHL and repression by tetR
Produces LuxR protein which can sense 3OC6HSL in the media and activate transcription from R0062, the right hand Lux promoter
Double terminator consisting of BBa_B0010 and BBa_B0012
V. fischeri <genbank>AF170104</genbank>
Synthesizes 3OC6HSL, which binds to LuxR. 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 repressor, LuxR. Two molecules of LuxR protein form a complex with two molecules the signalling compound HSL. This complex binds to a palindromic site on the promoter, increasing the rate of transcription.
An LVA tail (sequence: AANDENYALVA) was added to increase protein degradation. .
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr
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.
Conversion of the iGEM parts registry to SBOL2.1