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.permaflipped control for ZombiE.coli
| Types | DnaRegion |
| Roles | Plasmid plasmid |
| Sequences | BBa_K734006_sequence (Version 1) |
Description
This control plasmid is the ???permaflipped??? control, whose sequence is identical to that of the reporter plasmid but has the promoter flipped, as if already acted upon by Cre rcombinase. Using this control will show whether Z-Mode ("infected") cells actually express the RFP when Cre recombinase is assumed to work as intended, flipping the promoter region and generating one double-mutant and one wild-type loxP site. When transformed into cells and induced with IPTG, all cells should glow red if this control works correctly.Notes
In testing, induction with IPTG does not allow RFP 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 ampR rrnB terminator lasI strong RBS rfp strong RBS mutant loxP lacUV5 mutant loxP strong RBS sfGFP terminator ampR promoter rep_origin lacI | 1569,1589 2759,3619 2,159 190,795 796,806 837,1517 1518,1528 1529,1562 1563,1631 1632,1665 1666,1676 1677,2406 2437,2565 2689,2717 3774,4393 5808,6989 | operator feature/operator CDS feature/cds sequence_feature feature/misc CDS feature/cds ribosome_entry_site feature/rbs CDS feature/cds feature/rbs ribosome_entry_site feature/misc sequence_feature feature/promoter promoter sequence_feature feature/misc ribosome_entry_site feature/rbs CDS feature/cds sequence_feature feature/misc promoter feature/promoter sequence_feature feature/misc CDS feature/cds |