Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Naomi Genuth
Date created: 2011-09-20 11:00:00
Date modified: 2015-05-08 01:12:54
E. coli strain for His3-URA3 one-hybrid selection system
| Types | DnaRegion |
| Roles | chromosome Cell |
| Sequences | BBa_K615000_sequence (Version 1) |
Description
This strain is designed to test zinc finger binding using a one-hybrid metabolic selection system. The endogenous genes HisB and PyrF have been knocked out and replaced with the yeast versions His3 and URA3 under the control of a Zif268 binding site. The gene coding for the omega subunit of RNA polymerase (rpoZ) was knocked out by replacing it with a Zeocin cassette, and the strain is designed to be used in conjunction with BBa_K615001 (zinc finger-omega subunit expression construct).Notes
HisB and PyrF were knocked out using multiplex automated genome engineering (MAGE). Lambda red was employed to replace rpoZ with a zeocin cassette and to insert the kanamycin cassette-Zif268 binding site-His3-URA3 construct.Source
EcNR2 E. coli strain| Sequence Annotation | Location | Component / Role(s) |
| Kanamycin resistance cassette Target and promoter region Zif268 binding site His3 URA3 | 1,950 950,1080 978,986 1080,1742 1790,2593 | feature/misc sequence_feature feature/promoter promoter non_covalent_binding_site feature/binding CDS feature/protein feature/protein CDS |