AAVS1_Puro_PGK1_3xFLAG_Twin_Strep
(Plasmid #68375)

• Purpose: (Empty Backbone) Doyon lab Tandem-Affinity Purification Following (i) Nuclease-Driven Gene Addition to the AAVS1 Genomic Safe Harbor Locus and (ii) Endogenous TAP Tagging.
• 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: human PGK1
• Selectable markers: Puromycin
• Tags / Fusion Proteins:
  • 3xFLAG_Twin_Strep (N terminal on backbone)
  • 3xFLAG_Twin_Strep (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• 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_PGK1_3xFLAG_Twin_Strep_MCS
  • AAVS1_Puro_PGK1_3xFLAG_Twin_.gb
• Addgene Notes:
  • Addgene Diagnostic Digest
• 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:
  • 21 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• 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

Information for Cloning Grade DNA (Catalog # 68375-DNA.cg) ( Back to top)

Purpose

Cloning grade DNA is suitable for use in PCR, cloning reactions, or transformation into E. coli. The purity and amount is not suitable for direct transfections.

Delivery

• Amount: 2 µg
• Guaranteed Concentration: 100 ng/µl +/- 5 ng/µl
• Pricing: $105 USD
• Storage: DNA can be stored at 4℃ (short term) or -20℃ (long term).

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Quality Control

Addgene has verified this plasmid using Next Generation Sequencing. Results are available here