Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Ben Slater
Date created: 2012-09-30 11:00:00
Date modified: 2015-05-08 01:13:07
pReporter for ZombiE.coli
| Types | DnaRegion |
| Roles | plasmid Plasmid |
| Sequences | BBa_K734005_sequence (Version 1) |
Description
This plasmid contains a lacUV5 promoter between two mutant, inverted loxP sites (lox66 and lox77). When Cre recombinase is present, recombination flips the region between the loxP sites, causing the promoter to be oriented the opposite direction. This recombination should only be capable of occuring once, because after recombination one loxP site becomes wild-type and the other becomes double-mutant, disallowing binding by Cre.Before recombination, the promoter is oriented to express GFP, signaling healthy, "uninfected" cells. However, if pConverter expresses Cre (due to presence of the zombifying infectious agent AHL), the promoter is flipped to express RFP, signaling an "infected" cell. Downstream of rfp is lasI, which allows infected cells to have the ability to infect others.
Notes
In testing, induction with IPTG does not allow GFP expression, possibly due to extensive secondary structure on the RNA transcripts blocking the RBS.Source
LasI: from pSB1A2-K081009RFP: from pSB1AK3-K081014
double terminator: from pSB1AK3-B0015
backbone: pET21-derived
| Sequence Annotation | Location | Component / Role(s) |
| lacO lacUV5 ampR lacI ampR promoter rep_origin lox66 lox77 rrnB terminator terminator sfGFP rfp lasI strong RBS strong RBS strong RBS | 1605,1625 1563,1631 2759,3619 5808,6989 2689,2717 3774,4393 1529,1562 1632,1665 2,159 2437,2565 1677,2406 837,1517 190,795 1666,1676 1518,1528 796,806 | feature/operator operator feature/promoter promoter feature/cds CDS CDS feature/cds feature/promoter promoter feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature feature/misc sequence_feature CDS feature/cds CDS feature/cds CDS feature/cds feature/rbs ribosome_entry_site ribosome_entry_site feature/rbs ribosome_entry_site feature/rbs |