Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: David Caballero, Fernando Govantes
Date created: 2011-10-21 11:00:00
Date modified: 2015-05-08 01:12:30
pUC18Sfi-miniTn7BB-Gm-attTn7
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_K510047_sequence (Version 1) |
Description
This plasmid allows the integration of BioBricks into microbial chromosomes, using prefix or suffix cloning sites, without removing the attachment site of the Tn7 transposon (attTn7). This fact makes posible to integrate another BioBrick into the chromosome of a strain in which miniTn7BB-Gm-attTn7 transposition has occurred previously.Notes
In order to construct the pUC18Sfi-miniTn7BB-Gm delivery plasmid, the miniTn7BB-Gm transposon was cleaved from the commercial plasmid pMA (Mr. Gene) with SfiI and ligated to SfiI-digested pUC18Sfi. This strategy removes all multi-cloning restriction sites from pUC18Sfi, except for the flanking SfiI, thus guaranteeing that the unique sites in the transposon are not duplicated. The remove of the multi-cloning restriction sites was verified by analytic digestions. The direction of miniTn7BB-Gm insertion was determined by digestion.Source
The miniTn7BB-Gm minitransposon was synthesized commercially and pUC18SfiI is a commercial vector.The attTn7 was obtained from the purified genome of E. coli MC4100 by PCR (using a high accuracy Pfu polymerase) with the follow primers: gaattcgcggccgcttctagagTGCAGCTGCTGGCTTACCATG and ctgcagcggccgctactagtaACATGGAGTTGGCAGGATGTTTG.
| Access | Instance | Definition |
| public public | BBa_K510000 BBa_K510022 | BBa_K510000 BBa_K510022 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_K510000 BBa_K510022 | 1,4388 4397,4561 | BBa_K510000 BBa_K510022 |