pCB268
(Plasmid #53363)

• Purpose: pcDNA3.0-based plasmid encoding fDHFR-UbK48R-Aβ42 13myc under the control of T7 or CMV promoter for 35S-pulse-chase URT-based assays in rabbit reticulocyte extract.
• Depositing Lab: Alexander Varshavsky
• Publication: Brower et al Mol Cell. 2013 Apr 25;50(2):161-71. doi: 10.1016/j.molcel.2013.02.009. Epub 2013 Mar 14.

Backbone

• Vector backbone: pcDNA3.0 - Nco
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 6964
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: DHFR
• Species: M. musculus (mouse)
• Entrez Gene: Dhfr (a.k.a. 8430436I03Rik)
• Promoter: CMV
• Tag / Fusion Protein:
  • Flag (N terminal on insert)

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: n/a (unknown if destroyed)
• 5′ sequencing primer: CMV-F

Gene/Insert 2

• Gene/Insert name: Ub
• Species: H. sapiens (human)
• Tag / Fusion Protein:
  • HA (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: n/a

Gene/Insert 3

• Gene/Insert name: Aβ42
• Species: H. sapiens (human)
• Entrez Gene: APP (a.k.a. AAA, ABETA, ABPP, AD1, APPI, CTFgamma, CVAP, PN-II, PN2, alpha-sAPP, preA4)
• Tag / Fusion Protein:
  • 13xMyc (C terminal on insert)

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: n/a (unknown if destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: n/a
• 3′ sequencing primer: BGH-rev

Resource Information

• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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