AAVS1_Puro_Tet3G_3xFLAG_Twin_Strep
(Plasmid #92099)

• Purpose: (Empty Backbone) Doyon lab Tandem-Affinity Purification Following Nuclease-Driven Gene Addition to the AAVS1 Genomic Safe Harbor Locus. Transgene expression is controlled by an Auto-Regulated Tet-On 3G System.
• Depositing Lab: Yannick Doyon
• Publication: Dalvai et al Cell Rep. 2015 Oct 7. pii: S2211-1247(15)01020-7. doi: 10.1016/j.celrep.2015.09.009.

Backbone

• Vector backbone: pUC19
• Backbone size (bp): 2364
• Modifications to backbone: Insert was cloned into PvuII digested pUC19 (loss of 322bp including MCS)
• Vector type: Mammalian Expression, CRISPR, TALEN ; ZFN
• Promoter: Tet-On 3G Bidirectional
• Selectable markers: Puromycin

Growth in Bacteria

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

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: See protocol
• 3′ sequencing primer: See protocol

Resource Information

• Supplemental Documents:
  • AAVS1_Puro_Tet3G_FS_v2.gb
• A portion of this plasmid was derived from a plasmid made by: Modified from AAVS1 SA-2A-puro-pA donor (Plasmid #22075). New backbone, overall structure is similar but several modifications have been made through gene synthesis.
• Articles Citing this Plasmid:
  • 7 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Tet Systems Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

A detailed protocol for purification of protein complexes tagged with this vector can be found here: https://www.ncbi.nlm.nih.gov/pubmed/27372759