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URL: http://www.addgene.org/135656
Proper Citation: RRID:Addgene_135656
Insert Name: Intron containing lox site
Organism: Synthetic
Bacterial Resistance: Erythromycin
Defining Citation: PMID:31826971
Vector Backbone Description: Backbone Size:6500; Vector Backbone:pAT19; Vector Types:Bacterial Expression, Cre/Lox; Bacterial Resistance:Erythromycin
Comments: Use in combination with pQlox66F (addgene 135657) or pQlox 66R (addgene 135658), then pQcre1 (addgene 135659). Transcription of LtrB-derived group II intron is driven by the pFer promoter. Redesign intron to target gene using TargeTron or Clostron algorithms (links below), then clone by SOE-PCR or gene synthesis ligation between NdeI and BsrGI sites. Lox66 and Lox71 must be in same orientation to delete the intervening sequence, but the directions of introns relative to one another can generate inverted repeats in the chromosome after deletion has taken place, leading to futher recombinations events. See publication for details. Intron redesign links http://www.clostron.com/clostron1.php http://www.targetrons.com/targetron_pLtrB.php Reference for intron redesign Heap JT, Pennington OJ, Cartman ST, Carter GP, Minton NP. The ClosTron: A universal gene knock-out system for the genus Clostridium. J Microbiol Methods. 2007;70(3):452-464. Perutka J, Wang W, Goerlitz D, Lambowitz AM. Use of Computer-designed Group II Introns to Disrupt Escherichia coli DExH / D-box Protein and DNA Helicase Genes. 2004:421-439.
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Source: Addgene
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