pcDNA3.2-Cx43-ML-HA
(Plasmid #49857)

• Purpose: Encodes human connexin 43 with all internal methionines mutated to leucine and a C_terminal HA epitope fusion tag
• Depositing Lab: Robin Shaw
• Publication: Smyth et al Cell Rep. 2013 Nov 5. pii: S2211-1247(13)00578-0. doi: 10.1016/j.celrep.2013.10.009.

Backbone

• Vector backbone: pcDNA3.2/HA-DEST
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 7711
• Total vector size (bp): 8893
• Modifications to backbone: Replaced V5 c-terminal fusion tag with HA tag
• 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: Connexin 43
• Alt name: GJA1
• Species: H. sapiens (human)
• Insert Size (bp): 1146
• Mutation: All internal Methionines mutated to Leucine. Contains several wobble mutations to confer resistance to siRNA. No stop codon to create c-terminal fusion
• GenBank ID: NM_000165.3
• Entrez Gene: GJA1 (a.k.a. AVSD3, CMDR, CX43, EKVP, EKVP3, GJAL, HLHS1, HSS, ODDD, PPKCA)
• Promoter: CMV
• Tag / Fusion Protein:
  • HA (C terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: TK-pA-R

Terms and Licenses

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