Part:BBa_M36921:Design
Zinc Sensor
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 194
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30
Illegal PstI site found at 194 - 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 190
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 194
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 194
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Our genetic construct consists of a constitutive promoter (BBa_J23108) , a high-affinity ribosome binding site (BBa_J61101) , an optimized sequence of a ZntR regulatory gene (BBa_M36919) that controls the expression of the chromosomal zinc resistance operon znt , a transcriptional terminator consisting of a 64 base pair stem-loop (BBa_B0010) , and a znt promoter sequence with a palindromic binding site for ZntR (BBa_K190016) .
A promoter of medium strength (variant RFP 1303 au) was chosen because we wanted to not put a high translational burden on our cells while still being able to generate enough PoPS signal to guarantee expression of our selected Comet fluorescence output.
Source
All of our parts were pre-existing and sourced from the iGEM registry, making only slight modifications (silent mutations) to the original ZntR gene (BBa_M11082) in order to reduce the base pair lengths of palindromic sequences.
DNA2.0 also made modifications on the ZntR gene to optimize the sequence for E. coli.
References
Chaudhari, Aparna, and P. Babu. "Development of a Broad Spectrum Fluorescent Heavy Metal Bacterial Biosensor." PubFacts. PubMed, 7 Mar. 2012. Web. 20 Nov. 2014. <http://www.pubfacts.com/fulltext_frame.php?PMID=23070906&title=Development%20of%20a%20broad-spectrum%20fluorescent%20heavy%20metal%20bacterial%20biosensor>.