Ngn3 Cre-ER (DM#256)
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||15024||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
Vector backbonepBluescript II KS
- Backbone size w/o insert (bp) 3000
Vector typeMammalian Expression, Cre/Lox ; mouse transgenic
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Copy numberHigh Copy
Gene/Insert nameNgn3 promoter
SpeciesM. musculus (mouse)
Insert Size (bp)9000
Entrez GeneNeurog3 (a.k.a. Atoh5, Math4B, bHLHa7, ngn3)
/ Fusion Protein
- Cre-ER(TM) (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ sequencing primer T7
- 3′ sequencing primer T3 (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byCre-ER(TM) was a gift from Andy McMahon.
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:Ngn3 Cre-ER (DM#256) was a gift from Douglas Melton (Addgene plasmid # 15024 ; http://n2t.net/addgene:15024 ; RRID:Addgene_15024)
For your References section:Direct evidence for the pancreatic lineage: NGN3+ cells are islet progenitors and are distinct from duct progenitors. Gu G, Dubauskaite J, Melton DA. Development. 2002 May . 129(10):2447-57. PubMed 11973276
Map uploaded by the depositor.