pLKO.1 puro
(Plasmid #8453)

• Purpose: (Empty Backbone) 3rd gen lentiviral backbone for cloning and expression of new shRNA sequences. Uses puromycin for selection.
• Depositing Lab: Bob Weinberg
• Publication: Stewart et al RNA 2003 Apr;9(4):493-501.

Backbone

• Vector backbone: pLKO.1
• Backbone size (bp): 7032
• 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: None
• Alt name: LKO.1

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: LKO.1 5'

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest
• Reference:
  • Addgene's pLKO.1 protocol
• Articles Citing this Plasmid:
  • 401 References

Terms and Licenses

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

Depositor Comments

Empty lentiviral vector for siRNA expression; replaces Lentihair; see author's map. The 293T cell line can be obtained from the Weinberg lab or GenHunter http://genhunter.com/products/aptag-3/index.html

Note that Addgene's quality control sequence shows that there are about 25 nucleotides between the AgeI and EcoRI sites (the depositor sequences is not as accurate in this region).

For packaging, please use pCMV-dR8.2 dvpr (Addgene plasmid #8455) and pCMV-VSVG (Addgene plasmid #8454). For the official vector of The RNAi Consortium and a plasmid map, please see plasmid #10878.

Information for Cloning Grade DNA (Catalog # 8453-DNA.cg) ( Back to top)

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

• Amount: 2 µg
• Guaranteed Concentration: 100 ng/µl +/- 5 ng/µl
• Pricing: $105 USD
• Storage: DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

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

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here