pAAVS1-TLR targeting vector
(Plasmid #64215)

• Purpose: Targeting vector for the human AAVS1 locus for insertion of traffic light reporter (TLR) construct
• Depositing Lab: Ralf Kuehn
• Publication: Chu et al Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198.

Backbone

• Vector backbone: AAVS1 SA-2A-puro-pA donor
• Backbone manufacturer: Jaenisch lab (Addgene plasmid # 22075)
• Modifications to backbone: A CAG-Venus+1-P2A+3-tagRFP+3 cassette was cloned into SalI & NotI sites of AAVS1-SA-2A-Puro targeting vector (Addgene plasmid# 22075). The defective Venus (codons 117–152 replaced by a 52-bp segment derived from mouse Rosa26 locus) and a 56-bp segment from the mouse Rab38 gene were linked via T2A peptide to TagRFP in a 2-bp shifted reading frame (+3).
• Vector type: Human targeting
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: traffic light reporter (TLR)
• Alt name: AAVS1 targeting vector
• Alt name: pAAVS1-TLR donor
• Species: H. sapiens (human)
• Mutation: see publication for details
• GenBank ID: S51329.1
• Entrez Gene: AAVS1 (a.k.a. AAV)
• Promoter: CAG

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: M13pUC-rev
• 3′ sequencing primer: M13pUC-fwd

Resource Information

• Supplemental Documents:
  • Plasmid gb file
  • Plasmid sequence ingenbank format
• Articles Citing this Plasmid:
  • 8 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry