Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: David Caballero
Date created: 2011-09-18 11:00:00
Date modified: 2015-05-08 01:12:30
pUC18R6KT-miniTn7BB-Gm-RFP
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K510020_sequence (Version 1) |
Description
The mini-Tn7-Gm-RFP artificial transposon has been constructed based on pUC18R6KT-miniTn7-Gm (BBa_K510012) by inserting a RFP cassette (BBa_I13521) in its BioBrick cloning site (BCS). This plasmid can be used for visualization purposes or to brand a strain because of its insertion in the genome of the working organism and split the antibiotic resistance cassette with the temporary expression of Flp recombinase.Notes
The pUC18R6KT vector was amplified by PCR using as template a pTNS2 plasmid (GenBank accession number: AY884833) provided by Herbert P. Schweizer. The primers were designed to flanked the pUC18R6KT with SfiI restriction sites and allow the insertion of the mini-Tn7-Gm. Primers: attcGGCCTAGGCGGCCgtcgttttacaacgtcgtgac and attcGGCCGCCTAGGCCggaagcataaagtgtaaagcctg. The miniTn7BB-Gm minitransposon was synthesized commercially, and then digested at the flanking SfiI sites and cloned into SfiI-digested pUC18R6KT PCR products.Source
The BBa_I13521 BioBrick was inserted within the BCS of pUC18R6KT-miniTn7BB-Gm by EcoRI and PstI clonning.| Access | Instance | Definition |
| public public | BBa_K510012 BBa_I13521 | BBa_K510012 BBa_I13521 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K510012 BBa_I13521 | 1,4549 4558,5480 | BBa_K510012 BBa_I13521 |