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Addgene

AAV phSyn1(S)-FlpO-bGHpA
(Plasmid #51669)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 51669 Standard format: Plasmid sent in bacteria as agar stab 1 $85
AAV Retrograde 51669-AAVrg Virus (100 µL at titer ≥ 7×10¹² vg/mL) and Plasmid. $405

Backbone

  • Vector backbone
    pAAV-MCS
  • Backbone manufacturer
    Stratagene
  • Backbone size w/o insert (bp) 2887
  • Total vector size (bp) 4972
  • Modifications to backbone
    A fragment containing the hSyn1 promoter-FlpO recombinase gene-bGHpA were swapped in to replace the CMV promoter-MCS-hGHpA sequence. Total insert size including the two ITRs is 2375 bp
  • Vector type
    Mammalian Expression, AAV

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    30°C
  • Growth Strain(s)
    Stbl2
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    FlpO recombinase gene
  • Alt name
    Flp recombinase gene codon optimized for expression in mice.
  • Species
    S. cerevisiae (budding yeast)
  • Insert Size (bp)
    1299
  • Promoter hSyn1

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

  • A portion of this plasmid was derived from a plasmid made by
    Synthesized based on FlpO sequence in this reference: Raymond C.S. and P. Soriano.High-Efficiency FLP and ΦC31 Site-Specific Recombination in Mammalian Cells. 2007. PLoS One 2(1) e162.
  • Article Citing this Plasmid

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Information for AAV Retrograde (Catalog # 51669-AAVrg) ( Back to top)

Purpose

Ready-to-use AAV Retrograde particles produced from AAV phSyn1(S)-FlpO-bGHpA (#51669). In addition to the viral particles, you will also receive purified AAV phSyn1(S)-FlpO-bGHpA plasmid DNA.

hSyn-driven FlpO expression. 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

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

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

Retrograde functionality is dependent on high viral titers. Addgene recommends not diluting your AAV preps prior to use.
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)-FlpO-bGHpA was a gift from Hongkui Zeng (Addgene plasmid # 51669 ; http://n2t.net/addgene:51669 ; RRID:Addgene_51669) For viral preps, please replace (Addgene plasmid # 51669) in the above sentence with: (Addgene viral prep # 51669-AAVrg)
  • For your References section:

    Transgenic mice for intersectional targeting of neural sensors and effectors with high specificity and performance. Madisen L, Garner AR, Shimaoka D, Chuong AS, Klapoetke NC, Li L, van der Bourg A, Niino Y, Egolf L, Monetti C, Gu H, Mills M, Cheng A, Tasic B, Nguyen TN, Sunkin SM, Benucci A, Nagy A, Miyawaki A, Helmchen F, Empson RM, Knopfel T, Boyden ES, Reid RC, Carandini M, Zeng H. Neuron. 2015 Mar 4;85(5):942-58. doi: 10.1016/j.neuron.2015.02.022. 10.1016/j.neuron.2015.02.022 PubMed 25741722