hE-cadherin/α-catenin fusion-pcDNA3
(Plasmid #45771)

• Depositing Lab: Barry Gumbiner
• Publication: Gottardi et al J Cell Biol. 2001 May 28;153(5):1049-60.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5446
• Total vector size (bp): 10000
• 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): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: hE-cadherin/Xe α-catenin fusion
• Alt name: human E-cadherin
• Alt name: Xenopus α-catenin
• Species: H. sapiens (human), X. laevis (frog)
• Insert Size (bp): 4600
• Mutation: Human E-cadherin (aa 1-801) fused to Xenopus α-catenin (aa 201-end)
• GenBank ID: NM_004360.3; NP_004351.1 NM_001090631.1; NP_001084100.1
• Entrez Gene: CDH1 (a.k.a. Arc-1, BCDS1, CD324, CDHE, ECAD, LCAM, UVO)
• Entrez Gene: ctnna1 (a.k.a. cap102, catna1)
• Promoter: CMV

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: CMV-F; T7
• 3′ sequencing primer: BGH-rev; Sp6

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Human E-cadherin partial cDNA provided by David Rimm (Yale University, New Haven, CT)

Terms and Licenses

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

Depositor Comments

For this E-cadherin–α-catenin fusion construct that lacks the β-catenin binding domain in both E-cadherin and α-catenin, cDNA encoding the membrane proximal region of the cadherin cytoplasmic domain was amplified between nucleotides 1760 and 2530 using
5′ (5′-TGAGCACGTGAAGAACAGCACGTACAC-3′) and
3′ (5′-CCCTGGCCATTTCCAATTTCATCGGGA-3′)
oligonucleotide primers, and the resultant 770-bp fragment was cloned into a Xenopus α-catenin cDNA such that amino acid 809 in E-cadherin was fused in frame with amino acid 201 in α-catenin (E-cad-DEIGN/GHRDQ-α-catenin). This fusion region was further subcloned into the E-cad/pcDNA3 plasmid (Addgene plasmid 45769) to make the E-cadherin–α-catenin cDNA.