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PurposeExpresses an optimized mosquito OPN3 rhodopsin in-frame with mScarlet
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 125712 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 | |
AAV1 | 125712-AAV1 | Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid. | $405 | ||
AAV5 | 125712-AAV5 | Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid. | $405 |
Backbone
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Vector backbonepAAV
- Backbone size w/o insert (bp) 3788
- Total vector size (bp) 5609
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Vector typeMammalian Expression, AAV
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature30°C
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Growth Strain(s)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameeOPN3
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Alt nameMosquito OPN3, optimized for mammalian neuron expression
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gRNA/shRNA sequence-
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SpeciesAnopheles stephensi (Asian malaria mosquito)
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Mutationdeleted amino acids 331-429
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GenBank IDAB753162.1
- Promoter short version of αCamKinase II promoter (0.4kb)
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Tags
/ Fusion Proteins
- mScarlet (C terminal on insert)
- Rho1D4 (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 CKIIa_Fw (GATGCTGACGAAGGCTCGC)
- 3′ sequencing primer WPRE_Rev (CACACAGCGTAAAAGGAGCAAC) (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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Article Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Please visit https://www.biorxiv.org/content/10.1101/2021.02.18.431673v1 for the bioRxiv preprint.
Information for AAV1 (Catalog # 125712-AAV1) ( Back to top)
Purpose
Ready-to-use AAV1 particles produced from pAAV-CaMKIIa(0.4)-eOPN3-mScarlet-WPRE (#125712). In addition to the viral particles, you will also receive purified pAAV-CaMKIIa(0.4)-eOPN3-mScarlet-WPRE plasmid DNA.
CaMKIIa-driven expression of the optimized mosquito OPN3 rhodopsin in-frame with mScarlet. 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, AAV1 cap gene
- Buffer PBS + 0.001% Poloxamer 188 + 200 mM NaCl
- Serotype AAV1
- Purification Iodixanol gradient ultracentrifugation
- Reporter Gene mScarlet
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
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Viral 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 AAV5 (Catalog # 125712-AAV5) ( Back to top)
Purpose
Ready-to-use AAV5 particles produced from pAAV-CaMKIIa(0.4)-eOPN3-mScarlet-WPRE (#125712). In addition to the viral particles, you will also receive purified pAAV-CaMKIIa(0.4)-eOPN3-mScarlet-WPRE plasmid DNA.
CaMKIIa-driven expression of the optimized mosquito OPN3 rhodopsin in-frame with mScarlet. 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 mScarlet
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
-
Industry Terms
- Not Available to Industry
Viral 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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pAAV-CaMKIIa(0.4)-eOPN3-mScarlet-WPRE was a gift from Ofer Yizhar (Addgene plasmid # 125712 ; http://n2t.net/addgene:125712 ; RRID:Addgene_125712) For viral preps, please replace (Addgene plasmid # 125712) in the above sentence with: (Addgene viral prep # 125712-AAV1) or (Addgene viral prep # 125712-AAV5) -
For your References section:
Efficient optogenetic silencing of neurotransmitter release with a mosquito rhodopsin. Mahn M, Saraf-Sinik I, Patil P, Pulin M, Bitton E, Karalis N, Bruentgens F, Palgi S, Gat A, Dine J, Wietek J, Davidi I, Levy R, Litvin A, Zhou F, Sauter K, Soba P, Schmitz D, Luthi A, Rost BR, Wiegert JS, Yizhar O. Neuron. 2021 May 10. pii: S0896-6273(21)00161-6. doi: 10.1016/j.neuron.2021.03.013. 10.1016/j.neuron.2021.03.013 PubMed 33979634