pAAV-CBh-mKate2-IRES-MCS (C-gap-WT)
(Plasmid #105922)

• Purpose: AAV-mediated gene expression from CBh promoter. Contains left AAV2 ITR with an 11bp deletion in C region and canonical right AAV2 ITR
• Depositing Lab: Marcella Patrick
• Publication: AAV ITR constructs (unpublished)

Backbone

• Vector backbone: pAAV
• Modifications to backbone: 11bp deletion (aaagcccgggc) in left ITR removes 1 SmaI site
• Vector type: AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert 1

• Gene/Insert name: mKate2
• Mutation: codon optimized
• Promoter: CBh

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: SalI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: pCBhProF2 (5-agggtttaagggatggttgg)
• 3′ sequencing primer: pCDH-rev (5-GCATTCCTTTGGCGAGAG)

Gene/Insert 2

• Gene/Insert name: IRES-Multiple cloning site
• Alt name: MCS is FseI-PacI-SpeI-BamHI-XhoI-StuI-MluI-NheI-BmtI-AsiSI-AscI
• Promoter: same CBh promoter as mKate2

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: N/A (unknown if destroyed)
• 3′ cloning site: N/A (unknown if destroyed)
• 5′ sequencing primer: IRES-F (5-TGGCTCTCCTCAAGCGTATT)
• 3′ sequencing primer: WPRE-R (5-CATAGCGTAAAAGGAGCAACA)

Resource Information

• Supplemental Documents:
  • pAAV-CBh-mKate2-IRES-MCS Cgap WT final.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Industry MTA
  • Evrogen Ancillary Agreement for FP Distribution to Industry

Depositor Comments

Entire transgene cassette can be removed with NotI/XbaI digest; CBh promoter can be removed by XbaI/AgeI digest; mKate2 can be removed with SaI/EcoRI digest; IRES can be removed by EcoRI/FseI digest