pAAV-hSynapsin1-axon-GCaMP6s-P2A-mRuby3
(Plasmid #112005)

• Purpose: For enriched expression of GCaMP6s in axons with cytosolic expression of mRuby3
• Depositing Lab: Lin Tian
• Publication: Broussard et al Nat Neurosci. 2018 Sep;21(9):1272-1280. doi: 10.1038/s41593-018-0211-4. Epub 2018 Aug 20.

Backbone

• Vector backbone: pAAV
• Backbone size w/o insert (bp): 4603
• Total vector size (bp): 6787
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: axon-GCaMP6s-P2A-mRuby3
• Alt name: GAP43-GCaMP6s-P2A-mRuby3
• Species: Synthetic
• Insert Size (bp): 2184
• GenBank ID: MH282429
• Promoter: hSynapsin1
• Tag / Fusion Protein:
  • GAP43 palmitoylation domain (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: GAAGGCGCGCTGACGTCACTCGCCG
• 3′ sequencing primer: CAAATTTTGTAATCCAGAGGTTGATT

Resource Information

• Supplemental Documents:
  • pAAV-hSynapsin1-axon-GCaMP6s-P2A-mRuby3.gbk
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

Addgene NGS results found minor differences in the second ITR compared to the depositor-provided sequence. The depositing lab has confirmed that this does not impair AAV production

Information for AAV5 (Catalog # 112005-AAV5)

Purpose

Ready-to-use AAV5 particles produced from pAAV-hSynapsin1-axon-GCaMP6s-P2A-mRuby3 (#112005). In addition to the viral particles, you will also receive purified pAAV-hSynapsin1-axon-GCaMP6s-P2A-mRuby3 plasmid DNA.

Synapsin-driven, axon-targeted GCaMP6s calcium sensor and mRuby3 expression. The mRuby3 will be a separate protein (not a fusion protein). 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, AAV5 cap gene
• Buffer: PBS + 0.001% Poloxamer 188 + 200 mM NaCl
• Serotype: AAV5
• Purification: Iodixanol gradient ultracentrifugation
• Reporter Gene: mRuby3

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.

Information for AAV9 (Catalog # 112005-AAV9)

Purpose

Ready-to-use AAV9 particles produced from pAAV-hSynapsin1-axon-GCaMP6s-P2A-mRuby3 (#112005). In addition to the viral particles, you will also receive purified pAAV-hSynapsin1-axon-GCaMP6s-P2A-mRuby3 plasmid DNA.

Synapsin-driven, axon-targeted GCaMP6s calcium sensor and mRuby3 expression. The mRuby3 will be a separate protein (not a fusion protein). These AAV preparations are suitable purity for injection into animals.

Delivery

• Volume: 100 µL
• Titer: ≥ 1×10¹³ 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, AAV9 cap gene
• Buffer: PBS + 0.001% Poloxamer 188
• Serotype: AAV9
• Purification: Iodixanol gradient ultracentrifugation
• Reporter Gene: mRuby3

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:
  • Ancillary Agreement for Penn Vectors
  • 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.