AND GATE AraC+RBS(B0033)+T7ptag+Sal+SupD
It is made up of standard subparts from the partregistry.
{{PKU_Beijing2009_AND_GATE_FAMILY}} This is an AND GATE in Ecoli. Use salicylate to induce the transcription of SupD and use arabinose to induce the transcription of T7ptag. If the SupD tRNA exists, T7ptag can be translated into functional T7 polymerase, and otherwise the translation of T7ptag will come to stop because of some amber stop codon mutations in the T7ptag. The T7 polymerase will activate a T7 promoter. This gate is only one of nine gates(K228870-K228878), which are distinguished by the RBS of T7ptag. For more information: refer to K228000(T7ptag) and K228001(SupD) referrence: Anderson JC, Voigt CA, Arkin AP (2007) Environmental signal integration by a modular AND gate. Mol Syst Biol 3: 133
This is a salicylate inducible part and can produce supD, which is a tRNA coding gene and it can be well terminated by the terminator BBa_B0015.
from 2009 part registry distribution & Peking University 2009 igem team
T7 polymerase with an amber mutation coding gene is constructed after AraC protein(reversed sequence) and Pbad promoter which would be triggered by Arabinose. T7 polymerase can work well in the exsitence of SupD tRNA. Besides, the expression of the T7 polymerase can be regulated by RBS.
when cut this part for insert and ligate it into a vetor, the insert and vector may have the similar sizes which may disturb the ligation.
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.
Double terminator consisting of BBa_B0010 and BBa_B0012
RBS.4 (weaker) -- derivative of BBa_0030
Weaker RBS based on Ron Weiss thesis. Strengths relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-3" in figure 4-14 of thesis). 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.
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.
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 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.
This part could be connected to a protein coding sequence so that the expression could be induced by L-arabinose
Conversion of the iGEM parts registry to SBOL2.1