phage-cmv-cfp-24xms2
(Plasmid #40651)

• Depositing Lab: Robert Singer
• Publication: Wu et al Biophys J. 2012 Jun 20;102(12):2936-44. Epub 2012 Jun 19.

Backbone

• Vector backbone: pHAGE-CMV
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Growth instructions: Note it is possible that the 24x MS2 repeats may recombine even in Stbl3 cells. If this occurs, try growing the plasmid at 30C instead.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: CFP-24xMBS
• Alt name: 24 x MS2-binding sites (MBS)
• Species: Synthetic
• Insert Size (bp): 2054
• Promoter: CMV
• Tag / Fusion Protein:
  • CFP (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: WPRE-R (5'-CATAGCGTAAAAGGAGCAACA-3')

Resource Information

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

Terms and Licenses

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

Depositor Comments

The Singer Lab no longer recommends this plasmid, as it has a higher rate of bacterial recombination.

For mammalian cells they recommend the following plasmids:
pUbC-FLAG-24xSuntagV4-oxEBFP-AID-baUTR1-24xMS2V5-Wpre (Plasmid #84561)
HaloTag-bActinCDS-bActinUTR-MS2V5 (Plasmid #102718)

For yeast cells they recommend the following plasmids:
pET246-pUC57 12xMS2V6 (Plasmid #104390)
pET259-pUC57 24xMS2V6 (Plasmid #104391)
pET251-pUC 12xMS2V6 Loxp KANr Loxp (Plasmid #104392)
pET264-pUC 24xMS2V6 Loxp KANr Loxp (Plasmid #104393)

Due to the propensity of the plasmid to recombine, Addgene recommends screening multiple colonies by restriction analysis to ensure that the isolated clones contain a full set of 24 MS2 repeats