pET3a-SF-LMNC-TS
(Plasmid #169374)

• Purpose: E. coli expression plasmid for human LMNA residues 31-542, corresponding to the shorter lamin C isoform lacking the unstructured head, with an N-terminal splitFlAsH tag and a C-terminal TwinStrep tag.
• Depositing Lab: Bernd Bukau
• Publication: Bertolini et al Science. 2021 Jan 1;371(6524):57-64. doi: 10.1126/science.abc7151.

Backbone

• Vector backbone: pET3a
• Backbone size w/o insert (bp): 4600
• Total vector size (bp): 6365
• Modifications to backbone: NdeI + BamHI were used to open the MCS
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: LMNA residues 31-542
• Alt name: gene: LMNA
• Alt name: Protein isoform: Lamin C
• Species: H. sapiens (human)
• Insert Size (bp): 1752
• Mutation: deleted amino acid 1-30 (unstructured head domain), two silent mutations in the LMNA gene (isolated from U2OS cells) at plasmid position C4960T, G5157A
• GenBank ID: NM_001282625
• Entrez Gene: LMNA (a.k.a. CDCD1, CDDC, CMD1A, CMT2B1, EMD2, FPL, FPLD, FPLD2, HGPS, IDC, LDP1, LFP, LGMD1B, LMN1, LMNC, LMNL1, MADA, PRO1)
• Promoter: Native pET3a promotor
• Tags / Fusion Proteins:
  • Split-FlAsH Tag (N terminal on insert)
  • TwinStrep Tag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: T7 promotor fwd primer
• 3′ sequencing primer: T7 terminator rev primer

Terms and Licenses

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