T7 polymerase generator under TetR repressible promoter
This device is part of a filter system. When there is no TetR the key will be generated. These keys will then open the lock and this will lead to the production of T7 DNA polymerase. When TetR is present, there will be no production of T7 DNA polymerase.
FILTER This device was designed to filter out noise in an input signal; in our case OmpF PoPs. If there is a short increase in transcription from this promoter, a small amount of J23066 RiboKey is produced. This is then used to unlock (J23078) the short lived UmuD-tagged T7 RNA polymerase. This results in an increase of active T7 protein. The final part of this filter is an AND gate which is used in the components further on in the system. This AND gate consists of a T7 RNA polymerase promoter with another RiboLock (J23078 again). The AND gate will only function if the duration of the noise is sufficiently long and intense (= a signal) in order to have both the short-lived RiboKey AND T7 polymerase available at the same time and thus to produce the component.
We did a lot of modeling on this device. The most efficient Berkeley 2006 key and lock have been used. The RiboKy might be required in higher copy number although modeling doesn't predict this.
This part contains a T7 DNA polymerase under a constitutive promoter. The production of the polymerase is normally switched off because of the lock. It can only be switched on by the production of keys (see part BBa_J23066).
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
[key3c] riboregulator for lock3 variants
This part has the same homology region to J01122 as J23007 except that unlike J23007, J23008 does not contain the hairpin that was derived from J01129. J23008 anneals linearly to the stem of the hairpin in J01122 with perfect base pairing and destroys the hairpin in the process and reveals the RBS on J01122. When J01122 is transcribed the RNA secondary structure is such that the RBS hairpins with sequence upstream preventing the ribosome from binding and translation from occuring thereby locking up RFP further down stream. When J23008 is introduced into the cell in trans, we suspect that it will unlock the RBS for RFP as described above allowing translation to occur.
[key3d] riboregulator for lock3 variants
Variant of key3 (see part J01129 (key3), J23007(key3b), J23008(key3c)). Part is in pJ23006, so there is a Ptet upstream of the XbaI site to drive expression. Nevertheless, J23009 is still a basic part allowing both prefix and suffix insertions.
Double terminator consisting of BBa_B0010 and BBa_B0012
lock3d derivative with a 5' extension. This is the basic part, J23077 is the RFP reporter.
Fuse the LVA tag to the T7 coding region
Jonas Demeulemeester and Benjamien Moeyaert
Conversion of the iGEM parts registry to SBOL2.1