pMIR-Report-Luc-KLF4-K5S-Mt344A
(Plasmid #34601)

• Depositing Lab: Michael Ruppert
• Publication: Lin et al Mol Cell Biol. 2011 Jun;31(12):2513-27. Epub 2011 Apr 25.

Backbone

• Vector backbone: pMIR-Report-LUC
• Backbone manufacturer: Ambion
• Backbone size w/o insert (bp): 6470
• Vector type: Mammalian Expression, Luciferase
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL1 Blue
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: KLF4
• Alt name: GKLF
• Alt name: EZF
• Species: H. sapiens (human)
• Insert Size (bp): 306
• Mutation: The KLF4 K5S region was mutated at the miR-344 binding site: the seed binding region was mutated from CTGGATC to GAGGATC.
• GenBank ID: AF105036
• Entrez Gene: KLF4 (a.k.a. EZF, GKLF)
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: PmeI (destroyed during cloning)
• 3′ cloning site: PmeI (destroyed during cloning)
• 5′ sequencing primer: use KLF4 or firefly luciferase internal primers
• 3′ sequencing primer: use KLF4 or firefly luciferase internal primers

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This is a translation reporter. PCR primers were used to introduce mutations at the microRNA seed binding region of the K5S translational control element. The final constructs was confirmed by sequence analysis.