Pconst (strong)-luxR-Pconst (strong)-amilGFP-LVA
amilGFP(+LVA) generator(+RBS, +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.
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.
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr
Double terminator consisting of BBa_B0010 and BBa_B0012
amilGFP (+LVA), yellow reporter protein with degradation tag
PCR (template: BBa_K592010, primers shown on our wiki(UT-Tokyo 2013))
amilGFP (+LVA) yellow reporter protein with degradation tag. The strong yellow color is visible to the naked eye under natural room light both in a LB liquid medium and on a LB agar plate. You can easily visualize gene expression without special equipment. Rapid degradation of the protein allows real-time imaging . It is also useful as a fluorescent reporter. For detailed usage, see UT-Tokyo 2013 wiki. This is the LVA tagged version of BBa_K592010. LVA tag was added to increase decay rate by PCR using the primers shown on our wiki (UT-Tokyo 2013). amilGFP is a coral fluorescent protein from Acropora millepora. It is a member of the GFP family. Excitation maximum : 503 nm Emission maximum : 512 nm Quantum yield : 0.67 Molar Extinction : 75200 LVA tag is one of the ssrA degradation tag, which decreases the protein half-life when added to the C-terminus. References Andersen, J. B., Sternberg, C., Poulsen, L. K., Bj??rn, S. P., Givskov, M., & Molin, S. (1998). New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Applied and environmental microbiology, 64(6), 2240-2246. Labas, Y. A., Gurskaya, N. G., Yanushevich, Y. G., Fradkov, A. F., Lukyanov, K. A., Lukyanov, S. A., & Matz, M. V. (2002). Diversity and evolution of the green fluorescent protein family. Proceedings of the National Academy of Sciences, 99(7), 4256-4261.
LVA tag(11 aa) was added by PCR. As in the previous study(1). Additional 2 aa (Stu1 restriction site) was also added, which may not be necessary for unstabilization of the protein. RPAANDENYALVA double stop codon [1]Andersen, J. B., Sternberg, C., Poulsen, L. K., Bj??rn, S. P., Givskov, M., & Molin, S. (1998). New unstable variants of green fluorescent protein for studies of transient gene expression in bacteria. Applied and environmental microbiology, 64(6), 2240-2246.
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.
Conversion of the iGEM parts registry to SBOL2.1