Addgene - Hmga2 3'UTR m7 luciferase (Luc-m7) Plasmid Data

Plasmid 14788: Hmga2 3'UTR m7 luciferase (Luc-m7)

Gene/insert name:
Luc-m7
Alt name:
HMGA2
Insert size:
3000
Species:
M. musculus (mouse)
GenBank ID:
BC052158
Entrez Gene:
Hmga2 (9430083A20Rik, HMGI-C, Hmgic, pg, pygmy)
Fusion protein or tag:
luciferase
Terminal:
N terminal on backbone
Mutation:
m7 mutant (see article and author's map for details)
Vector backbone:
pIS1
(Search Vector Database)
Backbone manufacturer:
Available from Addgene (#12179)
Vector type:
Mammalian Expression, Luciferase
Backbone size w/o insert (bp):
4000
Cloning site 5':
XbaI
Site destroyed during cloning:
No
Cloning site 3':
NotI
Site destroyed during cloning:
No
5' sequencing primer:
N/A List of Sequencing Primers
3' sequencing primer:
EBV rev primer
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Sequence: View sequences (1)
Map:
View map
Principal Investigator:
David Bartel
Terms and Licenses:
MTA
Luciferase Limited Use Label License

Comments:

The 3' UTR of the mouse Hmga2 cDNA (BC052158) was subcloned into pCR2.1-TOPO (Invitrogen) for site-directed mutagenesis. To disrupt each let-7 complementary site, the nucleotides that paired to nucleotides 3 and 5 of the miRNA were substituted (see Author's map), using the Quikchange site-directed mutagenesis kit (Stratagene). To construct the Renilla luciferase reporters, wild-type and mutant Hmga2 3' UTRs were amplified (PCR primers, 5'-GCGTCTCGAGGGGCGCCGACATTC and 5'GGCGCGGCCGCAGTCAGAGGGCACAC) and cloned into the XbaI and NotI sites of pIS1.

Full sequence of wild-type plasmid available at Addgene plasmid #14785.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Disrupting the pairing between let-7 and Hmga2 enhances oncogenic transformation. Mayr et al (Science. 2007 Mar 16. 315(5818):1576-9. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 14788" in your Materials and Methods section.