Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Dylan Patrick Webster
Date created: 2012-07-25 11:00:00
Date modified: 2015-05-08 01:13:49
Arsenic inducible mtrB with cut sites flanking RBS
| Types | DnaRegion |
| Roles | Generator engineered_region |
| Sequences | BBa_K957000_sequence (Version 1) |
Description
The purpose of this part is to upregulate mtrB expression in response to arsenic. When expressed in a Shewanella strain lacking mtrB on the chromosome, this composite part will function as a component of a biosensor. Specifically, when such a complemented strain is inoculated in a microbial electrochemical system, current output will increase in response to arsenic.Within the genetic circuit, arsR acts as a negative autoregulator, repressing not only the expression of downstream mtrB, but also its own expression. Upon association with arsenic salts, ArsR dissociates from the operator of the arsenic inducible promoter, upregulating expression of downstream protein. Because mtrB activity is requisite for functionality of the mtr electron pathway in Shewanella spp., arsenic thus induces electron shuttling through the mtr pathway.
Notes
Primers were designed such that the wild-type Shine Dalgarno sequence was included upstream of the mtrB start codon, maintaining spacing. However, the appended BamHI cutsite downstream of the Shine Dalgarno sequence may alter translational efficiency.Source
Arsenic-sensitive component of this composite part was originally derived from the arsenic detoxification operon of E. coli strain JM109; BioBrick part BBa_J33201 was the source of DNA.mtrB is originally derived from the mtrCAB operon of Shewanella oneidensis MR-1; BioBrick part BBa_K098994 was used as DNA source.
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J33201 ArsR binding site -35 sequence -10 sequence rbs arsR AscI cut site mtrB rbs BamHI cut site mtrB BBa_K098989 KpnI cut site | 1,472 36,54 57,62 80,85 108,112 119,472 474,481 484,489 491,496 497,2590 497,2590 2620,2625 | engineered_region feature/BioBrick operator feature/operator promoter feature/promoter feature/promoter promoter feature/rbs ribosome_entry_site CDS feature/cds sequence_feature feature/misc ribosome_entry_site feature/rbs sequence_feature feature/misc feature/cds CDS feature/BioBrick engineered_region feature/misc sequence_feature |