pcDNA5-EGFP-AID-BubR1
(Plasmid #47330)

• Purpose: for plasmid integration using the Flp-In system. AID at N-terminal
• Depositing Lab: Don Cleveland
• Publication: Holland et al Proc Natl Acad Sci U S A. 2012 Dec 4;109(49):E3350-7. doi: 10.1073/pnas.1216880109. Epub 2012 Nov 12.

Backbone

• Vector backbone: pcDNA5/FRT/TO
• Backbone manufacturer: Invitrogen
• Total vector size (bp): 9850
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): XL10 Gold
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: BUB1 mitotic checkpoint serine/threonine kinase B
• Alt name: Bub1B
• Alt name: BubR1
• Species: H. sapiens (human)
• Insert Size (bp): 4590
• Mutation: siRNA resistant
• GenBank ID: NM_001211.5
• Entrez Gene: BUB1B (a.k.a. BUB1beta, BUBR1, Bub1A, MAD3L, MVA1, SSK1, hBUBR1)
• Tags / Fusion Proteins:
  • GFP (N terminal on insert)
  • AID (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: Not1 (not destroyed)
• 5′ sequencing primer: ACT AGT GGA TCC CTA TCA CTG AAA GAG CAA AGC CCC AGG ACT AGT TAA CT
• 3′ sequencing primer: ACT CGA GCG GCC GCC TAT CAC TGA AAG AGC AAA GCC CCA GGA CTA GTT AA

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest
• A portion of this plasmid was derived from a plasmid made by: The AID ORF was a kind gift from Masato Kanemaki (National Institute of Genetics, Mishima, Japan).
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Depositor Comments

Nishimura et al., 2009, Nature Methods