AraC generator then Lambda promoter controlling RFP production, "Scissors"
The AraC region comes from BBa_K228032. The promoter, rbs, rfp, and double terminator come from BBa_K1001754.
This is a project specific part. This sequence contains an AraC coding gene (BBa_K228032) upstream of a double terminator behind a Red Flourescent Protein and Ribosome Binding Site behind the modified Lambda Prm Promoter(BBa_K1001754). This specific modified Lambda Promoter is induced by the lamda cl repressor (BBa_C0051) instead of repressed by it. When induced, the RFP is expressed.
Terminator RFP and RBS behind Lamda Prm Promoter
This comes from BBa_K1001753 and BBa_B0015.
This sequence contains double terminator(BBa_B0015) behind a Red Flourescent Protein and Ribosome Binding Site behind the modified Lambda Prm Promoter(BBa_K1001753). This specific modified Lambda Promoter is induced by the lamda cl repressor (BBa_C0051) instead of repressed by it. When induced, the RFP is expressed. Use this BioBrick if your team wishes to have the inducible Lamda Prm promoter to control the expression of an RFP.
This uses the modified Lamda Prm Promoter specifically for its positive regulation.
<partinfo>BBa_J61127</partinfo> and <partinfo>C0080</partinfo>
Double terminator consisting of BBa_B0010 and BBa_B0012
araC arabinose operon regulatory protein (repressor/activator) from E. coli (+LVA)
GenBank: NC_002655 (www.ncbi.nlm.nih.gov)
coding region for the araC gene, used to make araC protein for use in positive or negative regulation of TIPs output (see also R0080, R0081)
We use the intermediate part, BBa_S03873, formed from an RFP (BBa_E1010) and an RBS (BBa_B0034).
This sequence contains a Red Flourescent Protein and Ribosome Binding Site (BBa_S03873) behind the modified Lambda Prm Promoter(BBa_I12006). This specific modified Lambda Promoter is induced by the lamda cl repressor (BBa_C0051) instead of repressed by it. When induced, the RFP is expressed. Use this BioBrick if your team wishes to have the inducible Lamda Prm promoter to control the expression of an RFP.
We specifically used the modified Lambda Prm Promoter because it was positively regulated.
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.
Modified lamdba Prm promoter (repressed by 434 cI)
Lamdba Prm promoter modified to be activated by lamda repressor (cI) and repressed by 434 repressor (cI)
The O-R1 region of 434 contained 14 base pairs as opposed to the 17 base pairs of the O-R3 site of lambda. Also, it was noticed that the O-R3 site of the lambda included part of the -10 site. Hence, to preserve the spacing and the -10 site, the three nucleotides that were in both the -10 site and the lambda O-R3 site were retained. The 14 nucleotides that were in the O-R3 site and not in the -10 site were replaced with the O-R1 site of the 434.
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.
**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.
Conversion of the iGEM parts registry to SBOL2.1