AND Gate pBad-SupD pSal-PhiR73 delta(amber mutation)
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.
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.
part registery 2009 distribution & PKU igem 2009 team
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.
part registery 2009 distribution & PKU 2009 igem team
The transcription of SupD-tRNA is regulated by the Pbad promoter(<partinfo>BBa_I13453</partinfo>). Arabinose can activate the transciprtion by binding to AraC regulatory protein.
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