|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||40904||Standard format: Plasmid sent in bacteria as agar stab||1||$85|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5400
- Total vector size (bp) 6800
Modifications to backboneFor the construction of the KLF5 plasmid pcDNA3-HA-KLF5-His, in which an HA tag was added to the N-terminus and a histidine tag was added to the C-terminus of KLF5, we first made the pcDNA3-HA-His vector by inserting a fragment of DNA containing an HA tag, EcoRI, XhoI, HindIII, BamHI, NotI, PstI and XbaI cloning sites and a histidine tag into the pcDNA3.1 vector (Invitrogen) between restriction enzyme sites NcoI and ApaI. The KLF5 cDNA was then cloned into this pcDNA3-HA-His vector between the HA and histidine tags at EcoRI and NotI sites using a PCR-based approach with the following primers (restriction enzyme sites are underlined): 5′-TTTGAATTCATGGCTACAAGGGTGC-3′ (forward) and 5′-TTTGCGGCCGCGTTCTGGTGCCTCTTCATA-3′ (reverse).
Vector typeMammalian Expression
Growth in Bacteria
Bacterial Resistance(s)Ampicillin, 100 μg/mL
Copy numberHigh Copy
Gene/Insert nameKruppel-like factor 5
SpeciesH. sapiens (human)
Insert Size (bp)1400
Entrez GeneKLF5 (a.k.a. BTEB2, CKLF, IKLF)
- Promoter CMV
/ Fusion Proteins
- HA (N terminal on insert)
- His (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site EcoR1 (destroyed during cloning)
- 3′ cloning site Not1 (destroyed during cloning)
- 5′ sequencing primer 5′-TTTGAATTCATGGCTACAAGGGTGC-3′
- 3′ sequencing primer 5′-TTTGCGGCCGCGTTCTGGTGCCTCTTCATA-3′ (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
- Zeocin® is an InvivoGen trademark.
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:pcDNA3-HA-KLF5-his was a gift from Jin-Tang Dong (Addgene plasmid # 40904 ; http://n2t.net/addgene:40904 ; RRID:Addgene_40904)
For your References section:Oestrogen causes degradation of KLF5 by inducing the E3 ubiquitin ligase EFP in ER-positive breast cancer cells. Zhao KW, Sikriwal D, Dong X, Guo P, Sun X, Dong JT. Biochem J. 2011 Jul 15;437(2):323-33. 10.1042/BJ20101388 PubMed 21542805