pRosetta
(Plasmid #59700)

• Purpose: Lentiviral vector for use as a standard for titering experiments when trying to compare vectors with different selection markers or to determine appropriate antibiotic conditions.
• Depositing Labs: John Doench, David Root
• Publication: David Root plasmids (unpublished)

Backbone

• Vector backbone: pLKO.1
• Total vector size (bp): 8765
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin, Blasticidin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: Puro
• Tag / Fusion Protein:
  • 2A (C terminal on insert)

Gene/Insert 2

• Gene/Insert name: BlastR
• Tag / Fusion Protein:
  • 2A (C terminal on insert)

Gene/Insert 3

• Gene/Insert name: EGFP

Resource Information

• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Depositor Comments

This lentiviral vector can be used as a standard for titering experiments when trying to compare vectors with different selection markers. It can also be used to determine if selection conditions with blasticidin or puromycin are using too high or too low a concentration of drug. If there are surviving cells that are EGFP-negative, then drug concentration is too low. If there are dying cells that express EGFP, then drug concentration is likely too high.