Axin DeltaGID/pCS2MT
(Plasmid #16303)

• Depositing Lab: Peter Klein
• Publication: Hedgepeth et al Mol Cell Biol. 1999 Oct . 19(10):7147-57.

Backbone

• Vector backbone: pCS2+MT
• Backbone size w/o insert (bp): 4352
• Vector type: Vertebrate expression

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: Axin DeltaGID
• Alt name: Axin
• Species: X. laevis (frog)
• Mutation: Delta GID: deletion of GSK interaction domain aa 324-504
• Entrez Gene: axin1.L (a.k.a. XELAEV_18045581mg, axin1, fused)
• Tag / Fusion Protein:
  • 5x myc (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: SP6
• 3′ sequencing primer: T3

Terms and Licenses

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

Depositor Comments

For sense transcripts, cut with NotI and transcribe with SP6.
GID= GSK3 interaction domain. NcoI digest should cut out a 1.733kb insert.

Please note that the GSK interaction domain deletion begins at aa 321 when the sequence is compared to GenBank entry NM_001088405.1. As a result of the cloning process, there are two amino acids 'HM' prior to the end of the deletion at aa 504 that are not part of the endogenous axin sequence.

Addgene's sequencing results also found a D559E mutation in the axin sequence that reflects a single nucleotide polymorphism. The depositing laboratory states that these differences are not a concern for the function of the plasmid.