pBOB-EF1-FastFUCCI-Puro
(Plasmid #86849)

• Purpose: 3rd generation lentiviral vector encoding FUCCI reporters for cell cycle analysis and puro for selection
• Depositing Labs: Kevin Brindle, Duncan Jodrell
• Publication: Koh et al J Cell Sci. 2016 Nov 25. pii: jcs.195164.

Backbone

• Vector backbone: pBOB
• Backbone manufacturer: Verma lab (Salk Institute)
• Backbone size w/o insert (bp): 7800
• Total vector size (bp): 12350
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: mKO2-hCDT1(30-120)
• Alt name: Fucci-G1 phase
• Species: Synthetic
• Insert Size (bp): 978
• GenBank ID: AB370332.1
• Promoter: EF1

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: PacI (not destroyed)
• 3′ cloning site: PacI (not destroyed)
• 5′ sequencing primer: TGATGTAATTCTCCTTGG

Gene/Insert 2

• Gene/Insert name: mAG-hGEM(1-110)
• Alt name: Fucci-S-G2-M phase
• Species: Synthetic
• Insert Size (bp): 1038
• GenBank ID: AB370333
• Promoter: EF1

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: PacI (not destroyed)
• 3′ cloning site: PacI (not destroyed)
• 5′ sequencing primer: AGAAGAGCACACCTGCCG

Gene/Insert 3

• Gene/Insert name: pac (puromycin N-acetyl-transferase)
• Alt name: puromycin resistance gene
• Species: S. alboniger
• Insert Size (bp): 600
• GenBank ID: M25346.1
• Promoter: PGK

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: none (destroyed during cloning)
• 5′ sequencing primer: ggcattctgcacgcttca

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 20 References

Terms and Licenses

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