FG12
(Plasmid #14884)

• Depositing Lab: David Baltimore
• Publication: Qin et al Proc Natl Acad Sci U S A. 2003 Jan 7. 100(1):183-8.

Backbone

• Vector backbone: HR'CS-G
• Backbone manufacturer: I. Verma, Salk
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: flap-Ub promoter-GFP-WRE

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: na

Resource Information

• Articles Citing this Plasmid:
  • 24 References

Terms and Licenses

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

Depositor Comments

FG12 was derived from FUGW. The extra nucleotides from the HindIII site downstream of the Ubiquitin-C promoter (UbiC) promoter to the NcoI site in front of the initiation codon of EGFP were deleted by a HindIII-NcoI adapter ligation. Further, XbaI, EcoRI, and XhoI sites at the 3' end of EGFP and WRE were eliminated, followed by a polylinker oligonucleotide ligation at the PacI site between the Flap element and the UbiC promoter, to generate a set of new restriction sites, XbaI-HpaI-XhoI-BstXI-PacI, that is optimal for accommodating the siRNA expression cassette. To construct the siRNA-expressing lentiviral vectors, the siRNA expression cassette can be subcloned into FG12 between the XbaI and XhoI sites.

Lentiviruses can be produced with third generation packaging system plasmids pMDLg/pRRE, pRSV-Rev, and pMD2.G.