Ngn3 Cre-ER (DM#256)
(Plasmid #15024)

• Depositing Lab: Douglas Melton
• Publication: Gu et al Development. 2002 May . 129(10):2447-57.

Backbone

• Vector backbone: pBluescript II KS
• Backbone size w/o insert (bp): 3000
• Vector type: Mammalian Expression, Cre/Lox ; mouse transgenic

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Ngn3 promoter
• Alt name: pGD29.1
• Species: M. musculus (mouse)
• Insert Size (bp): 9000
• Entrez Gene: Neurog3 (a.k.a. Atoh5, Math4B, bHLHa7, ngn3)
• Tag / Fusion Protein:
  • Cre-ER(TM) (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: T7
• 3′ sequencing primer: T3

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Cre-ER(TM) was a gift from Andy McMahon.

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor 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.