This system functions as a negative feedback to degrade extracellular insulin when extracellular glucose concentration is too low. It is essencial because when extracelluar glucose concentration is too low (hypoglycemia), subject death may occur.
Transcription terminator is added at the end so we can put system 1 and 2 in our project on the same vector.
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.
OmpR binding site. When position behind a promotor, it will turns that promotor into a OmpR controlled promotor. OmpR will function as an inhibitor of that promotor.
This part was derived from BBa_R0082 by PCR.
NCBI accession number: BC041675 K12 E. coli genomic DNA
Twin arginin signal peptide is an efficient protein export signal sequence of E. coli. The purpose of this construct is to allow E. coli export mouse insulin degradation enzyme(IDE) automatically, allowing controlled insulin degradation of extracellular insulin.
To combine TAT signal and mouse IDE, we used two separate PCR reaction to amplify each DNA fragment. We design the reverse primer of TAT signal and the forward primer of mouse IDE to be complement to each other, so when we mixed the product of each PCR reaction together, than used the forward primer of TAT signal and the reverse primer of mouse IDE to conduct PCR reaction, we can isolate the combine DNA product of TAT signal and mouse IDE. (Method devised by Dr. Chang)
unidentified/uncharacterized CinR dependent
Derived from luxICDABEG operon terminator of Vibrio fischeri <genbank>AF170104</genbank>.
Bidirectional transcriptional terminator consisting of a 22 bp stem-loop.
In the naturally-occuring sequence there is a mismatch in the stem of the stem loop. This can be corrected via an A->G mutation (at position 40 -- sequence coordinate/not MFOLD coordinate). The above sequence does not reflect this mutation (but the MFOLD image does). This terminator's location cannot be found using some inverted repeat detectors like PALINDROME because it is too short and contains a mismatch. This one was found with the help of Tom Knight. It lies between two coding regions that point towards eachother.
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.
Conversion of the iGEM parts registry to SBOL2.1