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pcDNA3-HA-KLF5-his
(Plasmid #40904)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 40904 Standard format: Plasmid sent in bacteria as agar stab 1 $94
DNA 40904-D50 50 μg of DNA in Tris buffer $495
DNA 40904-D100 100 μg of DNA in Tris buffer $585

Backbone

  • Vector backbone
    pcDNA3
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5400
  • Total vector size (bp) 6800
  • Modifications to backbone
    For 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 type
    Mammalian Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    Kruppel-like factor 5
  • Alt name
    KLF5
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    1400
  • GenBank ID
    NM_001730.3
  • Entrez Gene
    KLF5 (a.k.a. BTEB2, CKLF, IKLF)
  • Promoter CMV
  • Tags / Fusion Proteins
    • HA (N terminal on insert)
    • His (C terminal on insert)

Cloning Information

  • 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

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Trademarks:

  • Zeocin® is an InvivoGen trademark.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

How to cite this plasmid ( Back to top)

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