Locked RFP (J23077) with OriT (J01003). Lock that can be transferred via conjugation
J01003::J23077 this part and part J23121 should be able to conjugate into a donor cell and cause fluorescence. See parts J23077 and J23070 for explanation on locks and keys. a description of how to use part J01003 should be made and linked to from any J01003 part. hopefully before the jamboree!
see construction file on http://openwetware.org/wiki/IGEM:UC_Berkeley/2006 for actual construction method.
PCR ca1028F/ca1001 on pSB3C6-J23071 . . . (865 bp, XbaI/PstI) Sub into pJ23076 . . . . . . . . . . . . .(SpeI/PstI, large 2105) Product is pSB3C6-J23077, [Ptet][lock3I][RFP][TT]
lock3l with 5' extension and corrected RBS spacing
OriT-R (Origin of transfer for the R-plasmid nic region)
OriTR, the R plasmid nic region, is where the relaxosome nicks the plasmid and conjugative transfer by R plasmid machinery begins.
**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.
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
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.
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