This is a composite part formed from subparts available in the registry.
This construct is a scaffold device for measuring RBS strength via mCherry fluorescence. The device contains a ccdB expression cassette (BBa_P1010) in place of an RBS. Digestion with AvrII and SapI allows for the insertion of any RBS sequence via Heather Keller's alternative RBS assembly method, which allows for blunt cloning of an RBS upstream of a coding sequence.
Due to the presence of ccdB (BBa_P1010) this device can only be grown in DB3.1 or other permissive strains.
3' stabilizing mRNA designed by Christina Smolke and Jay Keasling for stabilizing gfp.
A single base pair change (A to T at position 25) from the original design was made in order to remove a PstI site occurring in the loop of the hairpin. This should have no effect on the secondary structure.
This part was generated by PCR amplification. The upstream biobricks cut sites (EcoRI and XbaI) as well as the R0011 promoter were incorporated into the forward promoter recognizing the ompA region in the MG1655 genome. The AvrII cut site, as well as the downstream biobricks cut sites (SpeI and PstI) were incorporated into the reverse promoter.
This part is an intermediate assembly product that contains the R0011 promoter (lambda cI promoter with lacI binding sites), the OmpaA 5' UTR containing a double hairpin for transcript stability, and an AvrII restriction site to facilitate later downstream cloning of an RBS via Heather Keller's RBS characterization scheme. These parts are cloned bluntly, without generating biobricks mixed sites in between.
This part was generated by PCR assembly in order to minimize the number of digestions and ligations necessary to generate a larger construct. Especially given the short sequence of the three individual parts (and the difficulting in purifying small parts) it was much simpler to amplify the product in this way rather than generate the 3 parts individually and clone them together.
The part was generated by PCR amplification of part E0040 with the SapI site incorported into the forward promoter.
mCherry part number J06504 with an upstream SapI site. Cleave with SapI results in an overhang of 3'-TAC - 5' from the left end of the mCherry coding region, making the coding sequence compatible with Heather Keller's alternative assembly method for blunt cloning of RBS sequences upstream of a coding sequence.
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.
The ccd operon having a constitutive promoter and the ccdB coding region. ccdB protein is lethal in normal cloning strains. This part is used to aid the process of moving a BioBrick part to a new plasmid. A target plasmid carrying P1010 is cut and mixed with the cut insert. Plasmids that recombine are selected against because they kill the cell that picks them up. P1010 is only provided as plasmid or in DB3.1 which is ccdB resistant.
Conversion of the iGEM parts registry to SBOL2.1