Strong promoter + Strong RBS + CH25H + 10x His tag + Double terminator
BBa_K880005 consists of strong promoter and strong rbs to maximize protein production. This construct codes for ch25h, a catalyst for the conversion of cholesterol to 25 hydroxycholesterol, and 10x histidine tag for protein purification. BBa__B0015 is a double terminator used to stop transcription.
We made sure that there was no Ecor1, Xbal, Spel1, and Pst1 cutting sites in the DNA.
Strong promoter, strong RBS combination for high expression levels of proteins
It is a combination of strong promoter (J23100) and RBS (B0034).
-J23100+B0034 -Strong promoter, strong RBS combination for high expression levels of proteins Consensus constitutive promoter and RBS sequence-produces strongest possible expression.
Josh Atkinson, Mike Ferguson, and Ben Parker
cDNA of ch25h was ordered from Sino BIological Inc
This DNA codes for ch25h, cholesterol 25-hydroxylase, that catalyzes the formation of 25 hydroxycholesterol from cholesterol. 25 hydroxycholesterol has been reported to have a range of effects on the immune system, including the suppression of production of IgA by B cells.
We removed three point mutations from the cDNA with mutagenesis done by MIssionBiotech.
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
10x-Histidine (10x-His) Tag with double stop codon (TAATAA)
Constructed through oligonucleotide annealing
10x-Histidine tag with double stop codon TAATAA to allow for better extraction of tagged products and protein termination in a single part.
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