pEGFP-C1 Utr230-EGFP-3XNLS
(Plasmid #58466)

• Purpose: Expresses a nuclear-targeted actin filament reporter, consisting of the first 230 amino acids of human utrophin, on a CMV promoter
• Depositing Lab: Dyche Mullins
• Publication: Belin et al Mol Biol Cell. 2013 Apr;24(7):982-94. doi: 10.1091/mbc.E12-09-0685. Epub 2013 Feb 27.

Backbone

• Vector backbone: pEGFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• Total vector size (bp): 5500
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: Utr230 with 3X glycine linker
• Species: H. sapiens (human)
• Insert Size (bp): 702
• GenBank ID: NM_007124
• Entrez Gene: UTRN (a.k.a. DMDL, DRP, DRP1)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (C terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: AgeI (not destroyed)
• 5′ sequencing primer: EGFP Reverse
• 3′ sequencing primer: CMV Forward

Gene/Insert 2

• Gene/Insert name: 3X NLS with 3X stop codons
• Species: SV40 virus
• Insert Size (bp): 72
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: KpnI (not destroyed)
• 5′ sequencing primer: EGFP Forward
• 3′ sequencing primer: SV40 polyA Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Utr230 was cloned from full-length human utrophin cDNA purchased from Open Biosystems (ThermoFisher). 3X NLS was cloned by insertion of annealed primers encoding 3xNLS (with stop codons) with flanking restriction sites.
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor Comments

Note: Plasmid contains a L221K mutation in EGFP. This mutation is not known to affect plasmid function.