Standard RBS expressing MS2 Coat protein ribozyme affinity purification (RAP)
This is an amalgamation of several biobricks an MS2 coat protein and the theophylline ribozyme (BBa_K1791000)
Standard RBS expressing MS2 Coat protein ribozyme affinity purification (RAP). This part utilizes the expression of C-terminal 6X His tagged fusion MS2 coat protein under a T7 promoter and a standard shine dalgarno sequence for expression of MS2. Downstream there is a T7 promoter followed by a theophylline ribozyme and a MS2 apatamer at the 3' end. Co-expression of the two modules results in MS2 binding to the 3' end of the theophylline ribozyme transcript allowing for the MS2-RNA purification by nickel affinity chromatography. The RNA can be eluted from the complex by the addition of theophylline.
We had to consider MS2 expression levels relative to the theophylline ribozyme transcript and assessed how this affects total RNA yield. In order to test this we developed a second part with depressed MS2 expression (BBa_K1791002)
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.
Double terminator consisting of BBa_B0010 and BBa_B0012
6X end terminally His tagged MS2 coat protein
MS2 coat protein can be easily purified via a nickel sepharose purification if this part is used downstream of a promoter and RBS.
T7 promoter fused with a theophylline inducible ribozyme (aptazyme)
Win, M.N., Smolke C.D. 2007 and BBa_I719005
This part was developed in order to increase the sequence space flexibility in dsRNA generated using BBa_K1791001 and BBa_K1791002
by changing the T7 promoter sequence (BBa_I719005) slightly after the GGG trinulceotide we get an increase in the potential for generating RNA with our composite parts BBa_K1791001 and BBa_K1791002.
This is an aptamer designed to bind MS2 coat protein upon transcription.
Had to test that it optimally binds to MS2 coat protein in vivo
This site will be utilized in the composite part BBa_K1791001 and BBa_K1791002 in order to allow for insertion of new RNA strands to be generated in these constructs.
no addtional design features were considered
no additional design considerations were neccessary
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