pcDNA3.4_CEACAM6-His
(Plasmid #149337)

• Purpose: for eukaryotic expression and purification of CEACAM6-His6
• Depositing Lab: James Fleckenstein
• Publication: Sheikh et al Proc Natl Acad Sci U S A. 2020 Nov 17;117(46):29055-29062. doi: 10.1073/pnas.2012480117. Epub 2020 Nov 2.

Backbone

• Vector backbone: pcDNA3.4_mIgG2-mMxr8a
• Backbone manufacturer: Diamond lab
• Backbone size w/o insert (bp): 7472
• Total vector size (bp): 6320
• Modifications to backbone: original plasmid digested with EcoRI/AgeI to remove portion of prior insert leaving IL-2 signal peptide sequence and C-terminal polyhistidine 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): DH10B
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: CEACAM6
• Species: H. sapiens (human)
• Insert Size (bp): 858
• Mutation: point mutation (GGA to GCA) to replace 'G-glycine' with 'A-alanine' at the site of lipidation required for GPI anchoring at plasmid position 1772
• Entrez Gene: CEACAM6 (a.k.a. CD66c, CEAL, NCA)
• Promoter: CMV
• Tags / Fusion Proteins:
  • polyhistidine (C terminal on insert)
  • IL-2 signal peptide (N terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TAAGTCTTGCACTTGTCACGAATTCGATATCGAAGCTCACTATTGAATCCACG
• 3′ sequencing primer: CTCAATGGTGATGGTGATGATGACCGGTTGCAGAGACTGTGATCATCGTG

Resource Information

• Supplemental Documents:
  • pCDNA3.4_CEACAM6-His.gb
• A portion of this plasmid was derived from a plasmid made by: original CEACAM6 clone was obtained from pRC202454 OriGene

Terms and Licenses

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