pRNA2-(A)128
(Plasmid #174006)

• Purpose: Used to generate stable mRNA for heterologous expression experiments (includes polyA tail)
• Depositing Lab: Stephen Ikeda
• Publication: Williams et al Front Neurosci. 2010 Nov 4;4:181. doi: 10.3389/fnins.2010.00181. eCollection 2010.

Backbone

• Vector backbone: pCI
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 4000
• Total vector size (bp): 5180
• Modifications to backbone: T7 promoter optimized and multiple cloning sequence heavily modified. Two copies of the human beta-globin 3'UTR were added. A 128bp polyA tail was added with downstream restriction enzyme cut sites. Ampicillin resistance gene was replaced with Kanamycin.
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: eGFP
• Species: Synthetic
• Insert Size (bp): 720
• GenBank ID: Cloned from U55762.1
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (destroyed during cloning)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: CMVfor

Terms and Licenses

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