GUM BUB3
(Plasmid #84029)

• Purpose: Expression of mCherry tagged BUB3 in human cells
• Depositing Lab: Patrick Paddison
• Publication: Toledo et al Dev Cell. 2014 Feb 10;28(3):282-94. doi: 10.1016/j.devcel.2013.12.014. Epub 2014 Jan 23.

Backbone

• Vector backbone: pGIPZ
• Backbone manufacturer: Open Biosystems
• Backbone size w/o insert (bp): 10381
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin and Bleocin (Zeocin), 100 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: BUB3
• Alt name: BUB3L
• Species: H. sapiens (human)
• Insert Size (bp): 2801
• GenBank ID: 9184
• Entrez Gene: BUB3 (a.k.a. BUB3L, hBUB3)
• Promoter: UCOE / SFFV
• Tag / Fusion Protein:
  • mCherry (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: mCherry-F
• 3′ sequencing primer: pCDH-rev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: BUB3 ORF from Open Biosystems

Terms and Licenses

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

Depositor Comments

Parental vector (pGIPZ containing a non-silencing control hairpin) was cut with XbaI + NotI to remove the CMV promoter and TurboGFP. UCOE / SFFV enhancer promoter complex and ORF with N-terminal mCherry were PCR amplified and Gibson cloned into the cut vector.