|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||25800||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4818
Vector typeMammalian Expression, Luciferase
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert namealternative hsa-miR-34a promoter
Alt namealternative pri-miR34a transcript
Insert Size (bp)2013
/ Fusion Protein
- luciferase (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site KpnI (not destroyed)
- 3′ cloning site NheI (not destroyed)
- 5′ sequencing primer n/a (Common Sequencing Primers)
promoter region of the alternative pri-miR-34a transcript identified by Navarro et al. Blood 2009 Sep 3;114(10):2181-92.
There are several discrepancies between this sequence and the GenBank sequence: T(at position 388) is cahnged to C
"ACACAC" insertion at position 658
"T" insertion at position 1914
A (at position 1599) changed to G
T (at position 1480) changed to G
G (at position 1212) changed to A
T (at position 1199) changed to C
These changes were present in all the clones that the lab sequenced and these clones came from two independent PCRs using genomic DNA extracted from the human erithroleukemia cell line K562. The changes might be due to that particular cell line. Importantly, all the minipreps I tested induced luciferase activity in K562 cells upon TPA treatment (as reported in the manuscript).
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:pGL3-PM-34a-K1/2 was a gift from Judy Lieberman (Addgene plasmid # 25800 ; http://n2t.net/addgene:25800 ; RRID:Addgene_25800)
For your References section:miR-34a contributes to megakaryocytic differentiation of K562 cells independently of p53. Navarro F, Gutman D, Meire E, Caceres M, Rigoutsos I, Bentwich Z, Lieberman J. Blood. 2009 Sep 3. 114(10):2181-92. 10.1182/blood-2009-02-205062 PubMed 19584398