ePB-PURO-FLAG-METTL3-APPA
(Plasmid #160253)

• Purpose: For stable integration of catalytic inactive FLAG-METTL3 in human cells with trasposase
• Depositing Lab: Alessandro Fatica
• Publication: Sorci et al Cell Death Dis. 2018 Jul 23;9(8):796. doi: 10.1038/s41419-018-0843-z.

Backbone

• Vector backbone: ePB-PURO
• Backbone size w/o insert (bp): 6956
• Total vector size (bp): 8600
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

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: methyltransferase like 3
• Alt name: METTL3
• Species: H. sapiens (human)
• Insert Size (bp): 1767
• Mutation: changes in aa 395–398, DPPW → APPA
• GenBank ID: NM_019852.5
• Entrez Gene: METTL3 (a.k.a. IME4, M6A, MT-A70, Spo8, hMETTL3)
• Promoter: tight TRE promoter
• Tag / Fusion Protein:
  • FLAG (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Hind III (not destroyed)
• 3′ cloning site: Not I (not destroyed)
• 5′ sequencing primer: CCCAAGCTTATGTCGGACACGTGGAGC
• 3′ sequencing primer: ATTTGCGGCCGCCTATAAATTCTTAGGTTTAGAGAT

Terms and Licenses

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