Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: John Chattaway
Date created: 2006-07-31 11:00:00
Date modified: 2015-08-31 04:08:48
Pops Blocker (Cre/Lox system)
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_J37027_sequence (Version 1) |
Description
This part is designed to be placed downstream of a promoter and prevent any Pops from the Promoter passing through this part. It will do this until an accompanying Cre Recombinase plasmid becomes activated. Once the Cre recombinase is activated the enzyme produced will permanently cut a section of DNA from the plasmid containing this part. This short section of DNA contains stop codons so once these are removed the polymerase can pass through this part and transcribe downstream genes. This short section of DNA is degraded.This is useful if a component of the system must be grown up but not activated until a certain external stimulus is added such as a positive feedback loop.
This part should be very efficient at preventing Pops passing through it.
The Part also contains a RFP reporter which is transcribed in the 3???-5??? direction. This means that un-activated parts will fluoresce red and activated parts will not fluoresce. This allows you to see that the part is working in your system. It also allows you to observe the efficiency of activation of the part in your system.
Because of the way Cre recombinase works the excised reporter will remain in a small plasmid and continue to be transcribed for a short time however this plasmid will not have an origin of replication so will not be copied and the fluorescent protein should stop being produced after around 15min
The design uses mutated lox sites, lox66 and lox71, which will make the excision irreversible.
Reference:
Zuwen Zhang and Beat Lutz (2002) Cre Recombinase-mediated inversion using lox66 and lox71: method to introduce conditional point mutations into the CREB-binding protein NUCLEAIC ACIDS RESEARCH
Notes
Used specific lox sites to make this irreversableSource
From Regestry:BBa_B0015
BBa_I13521
Lox Sites:
Zuwen Zhang and Beat Lutz (2002) Cre Recombinase-mediated inversion using lox66 and lox71: method to introduce conditional point mutations into the CREB-binding protein NUCLEAIC ACIDS RESEARCH
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J37026 BBa_B0010 stem_loop BBa_B0012 T7 TE polya stop BBa_J37028 Reversed and complementary to part I13521 | 1,34 37,116 48,91 125,165 132,151 152,165 158,158 1094,1127 168,1091 | feature/BioBrick engineered_region engineered_region feature/BioBrick stem_loop feature/stem_loop feature/BioBrick engineered_region feature/stem_loop stem_loop feature/polya polyA_site stop_codon feature/stop feature/BioBrick engineered_region feature/BioBrick engineered_region |