Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Eric Walters
Date created: 2011-09-22 11:00:00
Date modified: 2015-05-08 01:12:57
Double selection cassette (tagRFP-sacB-kanR)
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K634007_sequence (Version 1) |
Description
When placed 3' of the promoter portion of a 1- or 2-component sensor, BBa_634007 can be used in the following ways:Reporter: tagRFP (BBa_K634005) fluoresces red upon induction (see BBa_K634005 for issues with fluorescence)
Selection: Sensor systems can be selected for desired induced expression through supplementation of the culture with the sensor's analyte and kanamycin.
Counter selection: Sensors can also be selected for absence of expression when uninduced through supplementation of the culture with sucrose
Notes
We are still in the process of optimizing this device, the following considerations are undergoing testing:tagRFP was taken from an expression vector, did it lose 5' components essential for high expression?
Can we reduce the frequency of spontaneous loss of function mutations in sacB through protein fusion with kanR?
Is the kanR RBS too strong for the desired resolution of susceptibility? Can a weakened RBS increase the range of the selection?
Source
tagRFP came from EvrogensacB and kanR came from the registry
| Access | Instance | Definition |
| public public public | BBa_K634005 BBa_K322921 BBa_P1003 | BBa_K634005 sacB kanR |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K634005 BBa_K322921 BBa_P1003 | 1,750 759,2195 2204,3170 | BBa_K634005 sacB kanR |