Addgene - pIS1-mutant RhoA 3'UTR Plasmid Data

Plasmid 26090: pIS1-mutant RhoA 3'UTR

Gene/insert name:
mutant RhoA 3'UTR
Alt name:
mutant RhoA reporter
Alt name:
RhoA
Insert size:
1061
Species:
H. sapiens (human)
Entrez Gene:
RHOA (ARH12, ARHA, RHO12, RHOH12)
Mutation:
Both miR-31 binding sites in 3’ UTR mutagenized. Site 1: from ATCTTGC to AGGCGGC. Site 2: from TCTTGC to CGCCGC.
Vector backbone:
pIS1
(Search Vector Database)
Backbone manufacturer:
David Bartel, Addgene plasmid # 12179
Vector type:
Mammalian Expression, Luciferase
Backbone size w/o insert (bp):
4085
Cloning site 5':
SacI
Site destroyed during cloning:
Unknown
Cloning site 3':
AgeI
Site destroyed during cloning:
Unknown
5' sequencing primer:
Rluc-F (ccaggattcttttccaatgc) List of Sequencing Primers
3' sequencing primer:
EBV-rev
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Sequence: View sequences (1)
Principal Investigator:
Bob Weinberg
Terms and Licenses:
MTA

Comments:

Renilla luciferase reporter controlled by RhoA 3'UTR containing site-directed mutagenized miR-31 binding sites.

There is also a G->A mutation in the insert. The Weinberg lab has determined that this mutation does not affection function.

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

Article: A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis. Valastyan et al (Cell. 2009 Jun 12. 137(6):1032-46. 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 26090" in your Materials and Methods section.