Recombination Template for mamAB Operon with Promotor and Terminator
Magnetospirillum gryphiswaldense MSR-1 v2, complete genome
Consist of a template for recombination of mamAB operon(K1648000), a template for recombination of mamXY, mamGC and mms operon(K1648001), a promotor & RBS(J13002) in front of each template and a double terminator(B0015) at the back of each template. This part is designed for homologous recombination to introduce the large magnetosome forming operon into Azotobactor vinelandii. This part is introduced into A. vinelandii genome via random integration before full operon sequences to be transformed into A. vinelandii.
We add a promotor and RBS before each template to initial the coding and a double terminator(B0015) behind each template to terminate the coding and ligate these parts together.
Double terminator consisting of BBa_B0010 and BBa_B0012
This part is constructed by four fragments from the magnetospirillum gryphiswaldense magnetosome island, and it is to be used to do the homologous recombination with the genomic DNA of the magnetospirillum to form the whole coding sequence of
In order to form a homologous recombination template, we select 250 bp of each fragment and do the overlapping PCR ligate fragments 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.
Lutz, R., Bujard, H., Nucleic Acids Research (1997) 25, 1203-1210.
Sequence for pTet inverting regulator driven by the TetR protein.
BBa_R0040 TetR-Regulated Promoter is based on a cI promoter. It has been modified to include two TetR binding sites and the BioBrick standard assembly head and tail restriction sites.
June Rhee, Connie Tao, Ty Thomson, Louis Waldman
Conversion of the iGEM parts registry to SBOL2.1