pLD-hygro-EnVM
(Plasmid #24590)

• Depositing Lab: Jason Moffat
• Publication: Mak et al Mol Cell Proteomics. 2010 May;9(5):811-23.

Backbone

• Vector backbone: pLKO.1
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Hygromycin

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Ampicillin, 25 & 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): ccdB Survival
• Growth instructions: DB3.1 or ccdB resistance strain. We do not allow E. coli harbouring this plasmid to grow to saturation (no longer than 14 hours). We prefer to grow it at 30C and 180RPM of shaking when applicable.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Gateway(TM) cassette
• Insert Size (bp): 1705
• Tag / Fusion Protein:
  • VM (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: EF1-a

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

pLD-hygro-EnVM was constructed by subcloning the hygromycin resistance gene from the pLJM6 vector (J Moffat, unpublished) into the pLD-puro-EnFH vector using SpeI/NsiI and subsequently subcloning a gene synthesized V5-Myc (VM) tag (Bio Basic Inc.) from the pUC57 host vector.

pLD-puro-EnFH (pLD-puromycin resistance- EF1alpha, N-terminus triple Flag-6-His) was generated from pLKO.1 (see paper for details).