pLKO.1C
(Plasmid #139470)

• Purpose: Replacing the RSV promoter of pLKO.1 with the CMV promoter improves lentiviral production with third-generation viral packaging system.
• Depositing Lab: David Cobrinik
• Publication: Lee et al Biotechniques. 2020 Jun;68(6):349-352. doi: 10.2144/btn-2019-0155. Epub 2020 Mar 6.

Backbone

• Vector backbone: pLKO.1C
• Backbone size w/o insert (bp): 7442
• Modifications to backbone: RSV promoter of pLKO.1 was replaced with CMV promoter
• Vector type: Mammalian Expression, Lentiviral, RNAi
• 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

• Gene/Insert name: stuffer
• Insert Size (bp): 1900

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: TRC-F

Resource Information

• Supplemental Documents:
  • pLKO.1C.gbk
• A portion of this plasmid was derived from a plasmid made by: pLKO.1 was obtained from Addgene, and MA1 - where the CMV-HIV hybrid promoter was obtained - was kindly provided by Dr. Luigi Naldini of San Raffaele Telethon Institute for Gene Therapy,

Terms and Licenses

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

Depositor Comments

pLKO.1 : https://www.addgene.org/10878/
CMV-HIV hybrid promoter : Amendola M, Venneri MA, Biffi A, Vigna E, Naldini L. Coordinate dual-gene transgenesis by lentiviral vectors carrying synthetic bidirectional promoters. Nat. Biotechnol. 23 (1), 108-116 (2005).