CMV-Cx35-Halo-C-end
(Plasmid #71512)

• Purpose: Mammalian expression plasmid encoding perch Cx35 with a C-terminal HaloTag.
• Depositing Lab: John O'Brien
• Publication: Wang et al J Cell Sci. 2015 Nov 1;128(21):3888-97. doi: 10.1242/jcs.162586. Epub 2015 Sep 10.

Backbone

• Vector backbone: pEGFP-N1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 3961
• Total vector size (bp): 5937
• Modifications to backbone: EGFP removed; using backbone as CMV vector.
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Connexin 35
• Alt name: Cx35
• Alt name: Connexin 36
• Alt name: Cx36
• Species: Morone americana (Perch)
• Insert Size (bp): 2053
• GenBank ID: AF059183.1
• Tag / Fusion Protein:
  • HaloTag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: CMV forward
• 3′ sequencing primer: SV40 polyA

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • HaloTag Vector Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

C-terminal EGFP fusion on Cx35 replaced with HaloTag by cutting with SmaI and NotI and inserting HaloTag cut from pHaloTag-N with XhoI, ends filled with Klenow, and NotI.