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URL: http://www.addgene.org/15024
Proper Citation: RRID:Addgene_15024
Insert Name: Ngn3 promoter
Organism: Mus musculus
Bacterial Resistance: Ampicillin
Defining Citation: PMID:11973276
Vector Backbone Description: Backbone Size:3000; Vector Backbone:pBluescript II KS; Vector Types:Mammalian Expression, Cre/Lox, mouse transgenic; Bacterial Resistance:Ampicillin
Comments: The CRE-ER(TM) protein requires tamoxifen (TM) to catalyze LoxP site-mediated excision. To generate the Ngn3 constructs, a 7.6 kb fragment containing Ngn3 (pGD15, contains the 6.5 kb upstream region of Ngn3) was isolated using Ngn3 cDNA to screen a mouse RPI-22 BAC library (Genetics Institute, Buffalo, NY). Three primers were used to fuse the Cre-ERTM-coding region to the Ngn3 ATG to generate a fragment that had part of the Ngn3 promoter and the 5' end of the recombinase. The primers used are p73 (5'-acacttgactccttgatcgctg-3'), p71 (5'-gttgcatcgaccggtaatgca-3') and p74 (5'-ttttcccaaccgcaggatgtccaatttactgaccgtacac-3'). This fragment was subsequently digested with ApaI and AgeI and ligated to the KpnI (blunt-ended)-ApaI (partial digestion)-digested pGD15, and a 2.4 kb AgeI-SacII (partial digestion, blunt-ended) fragment of pGD35. The insert from the final construct (pGD29) was released by NotI digestion. See Author's Map and article for more information.
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Source: Addgene
