pDTA-5
(Plasmid #128845)

• Purpose: (Empty Backbone) Gateway-compatible p15A origin plasmid with a DTA negative selection cassette for targeting vector construction, I-CeuI, low copy number
• Depositing Lab: Mario Capecchi
• Publication: Additional gene targeting vector construction plasmids (unpublished)

Backbone

• Vector backbone: pWS-TK6
• Backbone manufacturer: Sen Wu, Capecchi laboratory
• Vector type: Mammalian Expression, Mouse Targeting

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DB3.1
• Copy number: Low Copy

Resource Information

• A portion of this plasmid was derived from a plasmid made by: DTA negative selection cassette is from Philippe Soriano. p15A origin backbone is from Sen Wu, Capecchi laboratory.

Terms and Licenses

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

Depositor Comments

Positive and negative selection enable efficient gene targeting in mouse embryonic stem cells.

A typical workflow is to subclone a genomic DNA fragment from a BAC clone using pStart-K #20346, then Gateway-clone (with LR Clonase II) the DTA negative selection cassette in this plasmid.

Different positive selection cassettes are available on Addgene, for example, pFNF #22687.

This plasmid is derived from pWS-TK6 #20350 and pSABgeolox2DTA #13447. The TK negative selection cassette was replaced by DTA negative selection cassette.