ccdA

BBa_K196000 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K196000
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Laetitia Warny
Date created: 2009-08-10 11:00:00
Date modified: 2015-05-08 01:11:17

CcdA antidote with the mob promoter (reverse)



Types
DnaRegion

Roles
Other

sequence_feature

Sequences BBa_K196000_sequence (Version 1)

Description

Stabilization system :
Higher plasmid stability = More proteins
Principle: In the StabyExpressTM system, the antidote gene (ccdA) is introduced in the plasmid DNA under the control of a weak constitutive promoter : the mob promoter, which doesn???t come from E. coli but from a broad host range plasmid (pBHR1) . On the other hand, the toxic gene (ccdB) is introduced in the chromosome of the bacteria, which can be furnished by DelphiGenetics. Expression of the poison gene is under the control of a promoter strongly repressed in the presence of the plasmid. When the plasmid is lost, the antidote is degraded and the production of the toxin is induced, causing cell death. Practically this means that when during the pre-induction phase bacteria are grown, 100% of the bacteria will carry the vector. If they lose the vector, they will not obtain a growth advantage, but will die. Upon induction 100% of the bacteria will start producing the recombinant protein leading to higher yields of the target protein and less background caused by unwanted proteins. For manufacturers of recombinant proteins this system offers a great benefit because it is an antibiotic free expression system. Therefore the manufactured protein will also be free of traces of antibiotics.
For more information, please visit www.delphigenetics.be.

Notes

Initially, the ccdA sequence was introduced in the plasmid in the reversed way. In order to leave the possibility to use it in both ways, we also designed this part in the forward way.

Source

This part comes from the StabyTM plasmid, designed by Delphi Genetics (www.delphigenetics.com).

Sequence Annotation Location Component / Role(s)
ccdA
mob (putative)
mob (putative)
putative RBS
1,219
323,328
303,308
227,231
feature/protein CDS
feature/promoter promoter
promoter feature/promoter
ribosome_entry_site feature/rbs
igem#experience
None
 
igem#sampleStatus
It's complicated
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K196000/1