pCol1a1-frt (FVB)
(Plasmid #63575)

• Purpose: Targeting vector for the Col1a1 locus that contains a Neo resistance cassette flanked by FRT sites, followed by an ATG-less Hygromycine resistance gene. The homology arms match the sequence of FVB.
• Depositing Lab: Ivo Huijbers
• Publication: Huijbers et al Nat Protoc. 2015 Nov;10(11):1755-85. doi: 10.1038/nprot.2015.114. Epub 2015 Oct 15.

Backbone

• Vector backbone: pBluescriptII SK
• Backbone size w/o insert (bp): 2888
• Total vector size (bp): 13163
• Modifications to backbone: restriction sites
• Vector type: Mammalian Expression, Bacterial Expression, Mouse Targeting ; FRT/Flpe
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Col1A-frt-hygro-pA
• Species: M. musculus (mouse)
• Insert Size (bp): 10275
• Promoter: None

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: SpeI (not destroyed)
• 5′ sequencing primer: M13reverse
• 3′ sequencing primer: T7

Resource Information

• Supplemental Documents:
  • pCol1a1-frt (FVB).gb
• A portion of this plasmid was derived from a plasmid made by: Rudolf Jaenisch lab

Terms and Licenses

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

Depositor Comments

The cassette in between the homologous recombination arms is the same as in AddGene Plasmid #20730.
The homologous recombination arms are derived from mouse strain FVB.
The Hygromycin gene in this construct does not have an ATG start codon nor is it expressed. Hygromycin can not be used for selection when this construct is used in a targeting experiment in mouse strain FVB derived ES cells.
The construct can be linearized with AscI, FseI or XhoI