pBabe RFP1-KRT5 hygro
(Plasmid #58493)

• Purpose: mammalian expression of mRFP1-tagged human KRT5
• Depositing Lab: Kevin Janes
• Publication: Wang et al Nat Cell Biol. 2014 Apr;16(4):345-56. doi: 10.1038/ncb2930. Epub 2014 Mar 23.

Backbone

• Vector backbone: pBabe N-terminal RFP hygro
• Backbone size w/o insert (bp): 6265
• Modifications to backbone: This construct is a variant of pBabe hygro that expresses mRFP1 as an N-terminal fusion. A new XhoI site was inserted immediately upstream of the BamHI site by PCR during cloning. TO EXPRESS AN N-mRFP1 FUSION, GENES MUST HAVE A BAMHI- OR XHO1-COMPATIBLE 5' STICKY END.
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: KRT5
• Alt name: keratin 5
• Species: H. sapiens (human)
• Insert Size (bp): 1783
• GenBank ID: NM_000424
• Entrez Gene: KRT5 (a.k.a. CK5, DDD, DDD1, EBS1, EBS2, EBS2A, EBS2B, EBS2C, EBS2D, EBS2E, EBS2F, K5, KRT5A)
• Tag / Fusion Protein:
  • mRFP1 (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: pBABE 5'
• 3′ sequencing primer: pBABE 3'

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

pBabe RFP1_KRT5 hygro was constructed by PCR cloning from plasmid templates (Open Biosystems) into the retroviral vector pBabe N-terminal RFP hygro. The PCR product was gel purified, digested with BamHI and EcoRI, and then ligated into pBabe N-terminal RFP hygro, which had been digested with BamHI and EcoRI and then dephosphorylated.