pcDNA3.1(+)mGAT1-0-GFP
(Plasmid #41662)

• Depositing Lab: Henry Lester
• Publication: Moss et al J Gen Physiol. 2009 Dec;134(6):489-521.

Backbone

• Vector backbone: pcDNA3.1(+)
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5428
• Total vector size (bp): 7899
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Mus musculus GABA transporter 1
• Alt name: mGAT1
• Alt name: sodium- and chloride-dependent GABA transporter 1
• Alt name: solute carrier family 6 member 1
• Species: M. musculus (mouse), Synthetic
• Insert Size (bp): 2549
• Mutation: C-terminal hydrophobic isoleucine residue added after GFP
• GenBank ID: M92378.1 AAB47998.1
• Entrez Gene: Slc6a1 (a.k.a. A730043E01, GABATHG, GABATR, GAT-1, Gabt, Gabt1, Gat1, XT-1, Xtrp1)
• Promoter: CMV
• Tag / Fusion Protein:
  • GFP (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: CMV-F; T7
• 3′ sequencing primer: GFPfusionRev; BGH_Rev

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

To fuse GFP to the C terminus of mGAT1, the mGAT1 open reading frame was subcloned into the HindIII and EcoRI sites of pcDNA3.1(+), and the GFP coding sequence was subcloned into the NotI and XbaI sites. A 12-residue spacer between mGAT1 and GFP was introduced by the multiple cloning site of the vector:

CGA ATT CTG CAG ATA TCC AGC ACA GTG GCG GCC GCC

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