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Plasmid Name
pMOD4-mT2A-mutated_rpsL-BSD-F
RRID:Addgene_182338 RRID Copied  
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RRID:Addgene_182338
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Plasmid Information

URL: http://www.addgene.org/182338

Proper Citation: RRID:Addgene_182338

Insert Name: BSD

Organism: Synthetic

Bacterial Resistance: Ampicillin

Vector Backbone Description: Backbone Marker:Alfred L. Fisher; Vector Backbone:pMOD4-galK-G; Vector Types:Bacterial Expression, Mouse Targeting, FLP / FRT; Bacterial Resistance:Ampicillin

Comments: PCR amplify around the 5' T2A and 3' FRT site, making sure to add a PacI site at the 3' of the FRT site. As suggested in the plasmid name, due to a PCR error, the rpsL open reading frame has a 1-bp deletion that results in a frame shift and as a consequence premature stop codons. Despite this, the downstream BSD open reading frame is translated from the bicistronic RNA in E. coli without problems. These mean that the mutated rpsL cannot be used for streptomycin counterselection, but the unmutated BSD can and was successfully used several times to confer blasticidin resistance for selection after recombineering.

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Data and Source Information

Source: Addgene