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AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE
(Plasmid #51509)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 51509 Standard format: Plasmid sent in bacteria as agar stab 1 $75
AAV1 51509-AAV1 Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid.
$380

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pAAV-MCS
  • Backbone manufacturer
    Stratagene
  • Total vector size (bp) 7634
  • Modifications to backbone
    A fragment containing the phSyn promoter-FLEX-tdTomato-T2A-synaptophysin fused EGFP-WPRE-bGHpA was swapped in to replace the CMV promoter-MCS-hGHpA sequence. Total insert size including the two ITRs is 5037 bp.
  • Vector type
    AAV

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    tdTomato-T2A-SypEGFP
  • Alt name
    tdTomato
  • Alt name
    EGFP
  • Alt name
    Synaptophysin
  • Insert Size (bp)
    1428
  • Promoter phSyn1

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Not1 (not destroyed)
  • 3′ cloning site Not1 (not destroyed)
  • 5′ sequencing primer none
  • 3′ sequencing primer none
  • (Common Sequencing Primers)

Resource Information

Information for AAV1 (Catalog # 51509-AAV1) ( Back to top )

Purpose

Ready-to-use AAV1 particles produced from AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE (#51509). In addition to the viral particles, you will also receive purified AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE plasmid DNA.

hSyn-driven, Cre-dependent expression of cytoplasmic tdTomato and presynaptic EGFP (synaptophysin-fused). These AAV preparations are suitable purity for injection into animals.

Delivery

  • Volume 100 µL
  • Titer ≥ 7×10¹² vg/mL
  • Pricing $350 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% Pluronic F-68 + 200 mM NaCl
  • Serotype AAV1
  • Purification Iodixanol gradient ultracentrifugation
  • Reporter Gene tdTomato (cytoplasmic, Cre-dependent), EGFP (presynaptic, Cre-dependent)

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

Resource Information

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

Using FLEX vectors in vivo: LoxP sites in FLEX plasmids are known to recombine during DNA amplification and viral vector production, which may result in a minority of Cre-activated (i.e., "flipped") viral vectors. Addgene has measured this occurs in 0.01-0.03% of viral vectors in our typical production protocol. This can lead to a small number of cells exhibiting Cre-independent transgene expression in vivo. To address this, we recommend titrating to find the optimal AAV dosage required for Cre-dependent transgene expression and function in vivo. This may include reducing the viral vector dosage in order to reduce the likelihood of Cre-independent expression.

How to cite this plasmid ( Back to top)

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.

  • For your Materials & Methods section:

    AAV phSyn1(S)-FLEX-tdTomato-T2A-SypEGFP-WPRE was a gift from Hongkui Zeng (Addgene plasmid # 51509 ; http://n2t.net/addgene:51509 ; RRID:Addgene_51509)

    For viral preps, please replace (Addgene plasmid # 51509) in the above sentence with: (Addgene viral prep # 51509-AAV1)

  • For your References section:

    A mesoscale connectome of the mouse brain. Oh SW, Harris JA, Ng L, Winslow B, Cain N, Mihalas S, Wang Q, Lau C, Kuan L, Henry AM, Mortrud MT, Ouellette B, Nguyen TN, Sorensen SA, Slaughterbeck CR, Wakeman W, Li Y, Feng D, Ho A, Nicholas E, Hirokawa KE, Bohn P, Joines KM, Peng H, Hawrylycz MJ, Phillips JW, Hohmann JG, Wohnoutka P, Gerfen CR, Koch C, Bernard A, Dang C, Jones AR, Zeng H. Nature. 2014 Apr 10;508(7495):207-14. doi: 10.1038/nature13186. Epub 2014 Apr 2. 10.1038/nature13186 PubMed 24695228