J23115 + RiboJ Promoter Characterization Part
Kalen Clifton, Christine Gao, Andrew Halleran, Ethan Jones, Likhitha Kolla, Joseph Maniaci, John Marken, John Mitchell, Callan Monette, Adam Reiss
UNS 2 Sequence, from Torella et al., 2013
Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, gkt860.
This is Unique Nucleotide Sequence 2, (UNS 2), from Torella et al., 2013. The William and Mary iGEM team has adopted this as our standard prefix; as such, all of our parts will have this sequence immediately following the BioBrick prefix. We took this measure in order to allow easier Gibson Assembly cloning of our parts. Primer sequences which can be used to clone with the UNS 2/3 standard can be found on our wiki.
UNS 2 was chosen because it works well with UNS 3 and it is in accordance with the BioBrick standard.
This is part B0034 with the scar region TACTAG added onto the part so that the correct scar region will be present in WM composite parts K2066059 - K20660108. This has no other purpose, but is just necessary do the system of the registry.
UNS 6.1 sequence derived from (Torella et. al 2013) "Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly".
This part is used as a spacer. WM iGEM 2016 used this part as a spacer to connect two separate composite parts onto the same plasmid. For example, UNS 6.1 was used as a spacer to connect K2066022(TetR on UNS) and K2066023(pTET GFP on UNS) to make K2066053.
Modified from the mCherry used in K2066028 and K2066029 to include the start codon. This modified mCherry is used in the construction of K2066040 and K2066041.
The start codon was removed in K2066028 and K2066029 to follow the design from Synthetic Analog Computation in Living Cells (Daniel et al.) where the codon bias was being optimized.
Pothoulakis et al. 2013 (???The Spinach RNA Aptamer as a Characterization Tool for Synthetic Biology???).
The spacer sB is taken from Pothoulakis et al. 2013 (???The Spinach RNA Aptamer as a Characterization Tool for Synthetic Biology???) and used in the construction of K2066040 and K2066041.
The broccoli sequence is from https://www.addgene.org/66843/.
This part is meant for use in Promoter Characterization and RBS Characterization subproject for WM iGEM 2016.
The part does have a SpeI cutsite in the Spinach aptamer. We???re going to use the original spinach to replicate the results of Pothoulakis et al. as exactly as possible and try to move to the Broccoli aptamer (which is RFC10 compatible) as soon as possible.
UNS 3 Sequence, from Torella et al., 2013
This is Unique Nucleotide Sequence 3, (UNS 3), from Torella et al., 2013. The William and Mary iGEM team has adopted this as our standard prefix; as such, all of our parts will have this sequence immediately following the BioBrick prefix. We took this measure in order to allow easier Gibson Assembly cloning of our parts. Primer sequences which can be used to clone with the UNS 2/3 standard can be found on our wiki. The sequence for this part came from the following paper: Torella, J. P., Boehm, C. R., Lienert, F., Chen, J. H., Way, J. C., & Silver, P. A. (2013). Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly. Nucleic acids research, gkt860. A huge thanks to all the researchers involved in its original creation!
This UNS sequence was chosen to serve as the 3' primer in our standard because it works well with UNS 2 and it adheres to the BioBrick standards.
Double terminator consisting of BBa_B0010 and BBa_B0012
Torella et al. 2013 (???Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly???)
The second half of UNS 6. The sequence is taken from Torella et al. 2013 (???Rapid construction of insulated genetic circuits via synthetic sequence-guided isothermal assembly???). This is used in K2066040 and K2066041.
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