pAAV-hsyn-Jaws-KGC-GFP-ER2
(Plasmid #65014)

• Purpose: AAV-mediated expression of Jaws under the human synapsin promoter
• Depositing Lab: Edward Boyden
• Publication: Chuong et al Nat Neurosci. 2014 Aug;17(8):1123-9. doi: 10.1038/nn.3752. Epub 2014 Jul 6.

Backbone

• Vector backbone: pAAV
• Backbone size w/o insert (bp): 4695
• Total vector size (bp): 6351
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl3
• Growth instructions: Please note that this backbone is highly recombination prone and should be amplified in a rec- strain like Sure2 or Stbl3, and grown at 30 C using carbenicillin rather than ampicillin.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Jaws-KGC-GFP-ER2
• Alt name: Jaws
• Species: Haloarcula salinarum (strain Shark)
• Insert Size (bp): 1650
• Mutation: K200R W214F
• GenBank ID: KM000926.1
• Promoter: human synapsin
• Tags / Fusion Proteins:
  • KGC (C terminal on insert)
  • GFP (C terminal on insert)
  • ER2 (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: EcoRI (not destroyed)
• 5′ sequencing primer: GCG AGG CGC GAG ATA G
• 3′ sequencing primer: CAA GGA GGA GAA AAT GAA AGC C

Resource Information

• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Information for AAV5 (Catalog # 65014-AAV5)

Purpose

Ready-to-use AAV5 particles produced from pAAV-hsyn-Jaws-KGC-GFP-ER2 (#65014). In addition to the viral particles, you will also receive purified pAAV-hsyn-Jaws-KGC-GFP-ER2 plasmid DNA.

hSyn-driven Jaws-GFP expression for optogenetic neural inhibition. 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
• Serotype: AAV5
• Purification: Iodixanol gradient ultracentrifugation
• Reporter Gene: GFP

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.

Addgene Comments

Data submitted about 65014-AAV5 by requesting scientist(s):

• Data 1: Rhesus macaque, Brain parenchyma

Information for AAV8 (Catalog # 65014-AAV8)

Purpose

Ready-to-use AAV8 particles produced from pAAV-hsyn-Jaws-KGC-GFP-ER2 (#65014). In addition to the viral particles, you will also receive purified pAAV-hsyn-Jaws-KGC-GFP-ER2 plasmid DNA.

hSyn-driven Jaws-GFP expression for optogenetic neural inhibition. 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, AAV8 cap gene
• Buffer: PBS + 0.001% Poloxamer 188
• Serotype: AAV8
• Purification: Iodixanol gradient ultracentrifugation
• Reporter Gene: GFP

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 AAV Retrograde (Catalog # 65014-AAVrg)

Purpose

Ready-to-use AAV Retrograde particles produced from pAAV-hsyn-Jaws-KGC-GFP-ER2 (#65014). In addition to the viral particles, you will also receive purified pAAV-hsyn-Jaws-KGC-GFP-ER2 plasmid DNA.

These AAV were produced with a retrograde serotype, which permits retrograde access to projection neurons. 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, AAV retrograde cap gene from rAAV2-retro helper (plasmid #81070)
• Buffer: PBS + 0.001% Poloxamer 188 + 200 mM NaCl
• Serotype: AAV retrograde (AAVrg)
• Purification: Iodixanol gradient ultracentrifugation
• Reporter Gene: GFP

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.

Addgene Comments

Data submitted about 65014-AAVrg by requesting scientist(s):

• Data 1: Rhesus macaque, Brain parenchyma