pAAV-CBh-mKate2-IRES-MCS (WT-WT)
(Plasmid #105921)

• Purpose: AAV-mediated gene expression from CBh promoter. Contains canonical left and right AAV2 ITRs
• Depositing Lab: Marcella Patrick
• Publication: AAV ITR constructs (unpublished)

Backbone

• Vector backbone: pAAV
• 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 WT-WT final.gb
• Articles Citing this Plasmid:
  • 2 References

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 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

Information for AAV2 (Catalog # 105921-AAV2)

Purpose

Ready-to-use AAV2 particles produced from pAAV-CBh-mKate2-IRES-MCS (WT-WT) (#105921). In addition to the viral particles, you will also receive purified pAAV-CBh-mKate2-IRES-MCS (WT-WT) plasmid DNA.

Control mKate2 expression AAV with two wild-type ITR sequences. These AAV preparations are suitable purity for injection into animals.

Delivery

• Volume: 100 µL
• Titer: ≥ 7×10¹² vg/mL
• Pricing: $375 USD for preparation of 100 µL virus + $30 USD for plasmid.
• Storage: Store at -80℃. Thaw just before use and keep on ice.
• Shipment: Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.

Viral Production & Use

• Packaging Plasmids: encode adenoviral helper sequences and AAV rep gene, AAV2 cap gene
• Buffer: PBS + 0.001% Poloxamer 188 + 200 mM NaCl
• Serotype: AAV2
• Purification: Iodixanol gradient ultracentrifugation
• Reporter Gene: mKate2

Biosafety

Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide

Terms and Licenses

• Academic/Nonprofit Terms:
  • Terms of Use for Viral Vectors
• Industry Terms:
  • Not Available to Industry

Viral Quality Control

Quality Control:

• Addgene ensures high quality viral vectors by optimizing and standardizing production protocols and performing rigorous quality control (QC) (see a list of our QC assays). The specific QC assays performed varies for each viral lot. To learn which specific QC assays were performed on your lot, please contact us.
• Titer: the exact titer of your sample will be reported on the tube. The titer you see listed on this page is the guaranteed minimum titer. See how titers are measured.

Visit our viral production page for more information.