pAJE-30-Ma-nitroY
(Plasmid #249487)

• Purpose: Expresses the Methanomethylophilus alvus pyrrolysine tRNA/tRNA synthetase pair engineered for 3-nitroTyr encoding in E. coli on a 30-copy pAJE origin, making it compatible with B95 cells.
• Depositing Lab: Ryan Mehl
• Publication: McGee et al Bio Protoc. 2026 Apr 20;16(8):e5674. doi: 10.21769/BioProtoc.5674. eCollection 2026 Apr 20.

Backbone

• Vector backbone: pAJE
• Backbone size w/o insert (bp): 3220
• Total vector size (bp): 4045
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: 3-nitrotyrosine RS (F5)
• Species: M. alvus
• Insert Size (bp): 825
• Mutation: N166A, V168S, A223C, W239R
• Promoter: GlnS

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: PstI (not destroyed)
• 5′ sequencing primer: N/A

Gene/Insert 2

• Gene/Insert name: Pyl-tRNA(6) (suppresses TAG codons)
• Species: M. alvus
• Insert Size (bp): 75
• Promoter: lpp

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: N/A

Terms and Licenses

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

Depositor Comments

3-nitrotyrosine RS (F5) insert is the same as Plasmid #225684, the difference being the low-copy pAJE origin (B95 cell compatible).