The pLac-RBS-T7RNAP gene came from geneart DNA synthesis. The pT7-RBS-GFP came from iGEM09_MoWestern_Davidson.
This gene was created to be a self promoter. The T7RNAP gene product is T7RNAP which binds to the pT7 promoter and cause the cell to fluoresce red.
The gene produces a red color but we have speculated that this is caused by read through transcription.
pLacI-RBS-T7RNAP with sites for oligo insertion
J23100 promoter and B0030 RBS upstream of T7 RNAP gene. The following restriction sites do not occur in the promoter-RBS-T7 RNAP sequence. Some of them did not occur in the original sequence and others were removed while conserving the amino acid sequence. Both the forward and reverse complement were checked for type IIs enzyme sites. EcoRI, XbaI, SpeI, PstI, AscI, AsiSI, BsaI, BsmBI, BbsI
BsmBI site (cgtctcn/ and /nnnnngagacg) before RBS and after gene coding sequence that can be used to cut out RBS-gene for placement after other promoters. Sites for AsiSI (gcgat/cgc) and AscI (gg/cgcgcc) that can be used to insert existing oligos. Sites flanking these for BsaI (ggtctcn/ and /nnnnngagacc) that can be used to insert new oligos without the need for gel purification. A site for BbsI (gaagacnn/ and /nnnnnngtcttc) that produces a sticky end complementary to the first BsaI site, for use in generating a near wild type version of the gene.
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.
The T7 promoter should only produce PoPS when the T7 polymerase is also being expressed.
Conversion of the iGEM parts registry to SBOL2.1