Built by joining different standardized biobricks together
This is a branch of the AND GATE built by Peking University 2009 iGEM Team. The addition of arabinose 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.
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.
The part is RBS->T7ptag->Terminator. K228002 is added to the RBS BBa_B0030 to construct this part. 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.
Because the RBS is only about 10bp, so let the BBa_K228002 be the insert and the backbone with RBS be the vector would be more convenient.
RBS.1 (strong) -- modified from R. Weiss
Strong RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("orig" in figure 4-14 of Ron Weiss 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 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)
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