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

• Depositing Lab: Jin-Tang Dong
• Publication: Zhao et al Biochem J. 2011 Jul 15;437(2):323-33.

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′

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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