AND GATE AraC+SupD+Sal+RBS(B0034)+T7ptag
Built by joining different standardized biobricks together
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.
AraC protein(reversed sequence) and Pbad promoter
This part contains AraC protein generator(BBa_K228008) and Pbad promoter(BBa_I13453). The expression of any coding sequence following the Pbad promoter is repressed constitutively by AraC protein and induced by L-arabinose.
The maximum concentration of L-arabinose we tested is 10^-4 M.
It is made up of the basic subparts from the partsregistry.
SupD is a tRNA coding gene, and it can be well terminated by the terminator BBa_B0015. You can fuse them after a certain promoter.
This is a branch of the AND GATE built by Peking University 2009 iGEM Team. The addition of salicylate can induce the transcription of T7ptag. However, if the SupD tRNA is absent, T7ptag might not be translated into functional T7 polymerase normally because of some amber stop codon mutations in it. We also have constructed a few similar parts with various RBS, which would make the expression regulation of T7ptag gene more convenient by choosing different parts.
T7ptag(T7polymerase with amber mutation)
The T7ptag is standardlized from the plasmid kindly provided by Voigt Lab.
T7ptag is a coding sequence of T7 polymerase with 2 Amber mutations. The transcription of T7ptag gene can only lead to the generation of its mRNA, further translation into T7 RNA polymerase is blocked because of the amber mutation. Only in the presence of the Amber mutation suppressor BBa_K228001 SupD-tRNA, it can be translated into functional T7 polymerase. T7ptag is a component of the AND Gate constructed by the Voigt Lab(UCSF)
SupD-tRNA again is from Voigt Lab(UCSF), We standardlize this part by PCR strategy.
The SupD-tRNA part functions as a tRNA, it suppresses the TAG amber stop codon and translate it into a Ser. There is no need to assembly SupD-tRNA with a rbs, since it is not translated. This part, together with the T7ptag part, make an AND Gate, that is, in the presence of both the 2 parts, T7 polymerase is tranlated into a functional polymerase that transcrips a downstream element constructed after a T7 promoter. Which is the output of the AND Gate.
There is a PstI site in the original sequence, but luckily it is near the 3' end of this part, so we designed a mutation in our primer and mutated the PstI site.
This part could be connected to a protein coding sequence so that the expression could be induced by L-arabinose
It is constrcuted by PCR from the plasmid kindly provided by Voigt Lab in UCSF.
This is a salicylate inducible promoter part. The activator protein is right upstream but in a opposite direction of the regulated promoter. In the presence of salicylate, the promoter can be activated.
There is no termoinator downstream of the NahR coding sequence, but it has no effect on parts that are in the same direction as the pSal promoter. However, if coding gene is placed upstream of this part and in the same direction as NahR, it may cause unwanted expression.
Conversion of the iGEM parts registry to SBOL2.1