PurposeUsed to sense miRNA activity using fluorescence based tools. hsa-miR-146a-5p target in 3' UTR of of mKate. miRNA activity can be measured by the repression of mKate2 relative to EBFP2 fluorescent proteins
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||103248||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Vector backboneLow Sensor Backbone (LSB)
Vector typeMammalian Expression, Synthetic Biology
Growth in Bacteria
Bacterial Resistance(s)Ampicillin and Kanamycin
Growth Strain(s)NEB Stable
Copy numberHigh Copy
Gene/Insert namehsa-miR-146a-5p target
SpeciesH. sapiens (human)
- Promoter EF-1a
- Cloning method Gibson Cloning
Please note that this plasmid is inherently unstable due to a repeating poly A element. We recommend performing a diagnostic digest on multiple colonies upon receipt. The depositors would like to note the presence of several discrepancies between the depositor sequence and physical plasmid sequence. In particular, the sequences upstream and downstream of the puromycin coding region have 10 bp insertions which do not affect the ability to perform puromycin selection. The sequences for those regions within the physical plasmids are as follows: Upstream of Puro: GAATTCGACCGCCTGTCTCAAGGTGCCACC; Downstream of Puro: GCTTTGAGACTACGCGTGAATTCACTCCTC.
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:LSB-hsa-miR-146a-5p was a gift from Ron Weiss (Addgene plasmid # 103248 ; http://n2t.net/addgene:103248 ; RRID:Addgene_103248)
For your References section:A mixed antagonistic/synergistic miRNA repression model enables accurate predictions of multi-input miRNA sensor activity. Gam JJ, Babb J, Weiss R. Nat Commun. 2018 Jun 22;9(1):2430. doi: 10.1038/s41467-018-04575-0. 10.1038/s41467-018-04575-0 PubMed 29934631