pTet Regulated Arg-tagged ECFP and EYFP (FRET Reporter)
[http://partsregistry.org/Part:BBa_K331033 BBa_K331033] and [http://partsregistry.org/Part:BBa_K331035 BBa_K331035] were both assembled by the Lethbridge 2010 team.
Made by assembling [http://partsregistry.org/Part:BBa_K331033 BBa_K331033] with [http://partsregistry.org/Part:BBa_K331035 BBa_K331035]. The pTet promoter is constitutively "on"; thus, Arg-tagged ECFP and Arg-tagged EYFP will be produced constitutively. By placing this part downstream of [http://partsregistry.org/wiki/index.php?title=Part:BBa_K542003 BBa_K542003], expression of the fluorescence proteins maybe "turned off" in the presence of arabinose. Fluorescence/F??rster Resonance Energy Transfer(FRET) is a distance-dependent phenomenon in which the excitation of a donor fluorophore leads to emission by an acceptor fluorophore; this occurs if the two fluorophores are within a certain distance to each other. The distance is dependent on the FRET pair used. The emission spectrum of the donor fluorophore must overlap with the excitation spectrum of the acceptor fluorophore for FRET to occur. FRET is explained in further detail on the [http://2009.igem.org/Team:Lethbridge/Project#FRET Lethbridge 2009 Wiki]. This phenomenon will allow for characterizing co-localization within the Lumazine Synthase microcompartment.
In order to characterize the Lumazine Synthase microcompartment, the fluorescent proteins must be regulated by a different promoter than that of Lumazine.
Tet Repressible C-terminal Arg Tagged ECFP (no terminator)
This is <partinfo>BBa_K331025</partinfo> with the tetracycline repressible promoter <partinfo>BBa_R0040</partinfo> added in front. This will be a component of a proof-of-concept part that will show localization of proteins into the interior of a microcompartment formed by the assembly of lumazine synthase proteins.
C-terminal Arg Tagged EYFP with transcriptional terminator
This is <partinfo>BBa_K331023</partinfo> with a transcriptional terminator <partinfo>BBa_B0015</partinfo> added behind. This will be a component of a proof-of-concept part that will show localization of proteins into the interior of a microcompartment formed by the assembly of lumazine synthase proteins.
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.
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
Fusion EYFP with C-terminal Arginine Tag
BioBrick Foundation EYFP fused with a synthesized Arginine tag
This Yellow Fluorescent protein has been fused with 10 C-terminal Arginines to target it into the Lumazine-based microcompartment.
The idea came from: Seebeck, F. P., Woycechowsky, K. J., Zhuang, W., Rabe, J. P., and Hilvert, D., (2006). A simple tagging system for protein encapulation. J. Am. Chem. Soc. 128, 4516-4517
This protein is the ECFP fused with the C-terminal Arginine tag found in BBa_K249005. It is designed to be part of a construct for targeting into the lumazine synthase microcompatment (BBa_K331000)
Conversion of the iGEM parts registry to SBOL2.1