ssAAV-EF1a-FLuc-WPRE-HgHpA_bac_293
(Plasmid #118412)

• Purpose: Expresses firefly luciferase under the control of a strong ubiquitous promoter EF1a along with a WPRE cassette. Backbone is compatible with both baculoviral and human production methods of AAV.
• Depositing Lab: Nicole Paulk
• Publication: Rumachik et al Mol Ther Methods Clin Dev. 2020 May 22;18:98-118. doi: 10.1016/j.omtm.2020.05.018. eCollection 2020 Sep 11.

Backbone

• Vector backbone: pFB-AAV-mcs-shuttle_UIowa G0202
• Backbone manufacturer: University of Iowa
• Backbone size w/o insert (bp): 7290
• Total vector size (bp): 8943
• Modifications to backbone: Has both the Tn7/GmR components for making a baculovirus expressing AAV, as well as the AmpR components for use when producing plasmid in E.coli for use when making AAV in human HEK293(T) cells
• Vector type: Mammalian Expression, Insect Expression, AAV, Luciferase, Synthetic Biology
• Selectable markers: Gentamicin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Growth instructions: Must check ITR integrity by restriction digest with AhdI, as well as with XmaI, to ensure the vector hasn't recombined following amplification.
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Firefly luciferase
• Alt name: FLuc
• Species: Photinus pyralis
• Insert Size (bp): 1653
• Promoter: EF1a

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: TCAGACAGTGGTTCAAAG
• 3′ sequencing primer: CATTAAAGCAGCGTATCC

Resource Information

• Supplemental Documents:
  • ssAAV-EF1a-FLuc-WPRE-HgHpA_bac_293.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry