pAAV-ITR-PytR(PyOtR)-mCherry
(Plasmid #204870)

• Purpose: Expresses pyrrolysyl tRNA variant "PyOtR" and a wild-type mCherry reporter; can be packaged into AAV
• Depositing Lab: Abhishek Chatterjee
• Publication: Jewel et al Angew Chem Int Ed Engl. 2024 Jan 26:e202316428. doi: 10.1002/anie.202316428.

Backbone

• Vector backbone: pAAV
• Backbone manufacturer: Cell Biolabs
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 5849
• Modifications to backbone: Replaced GFP with mCherry fluorescent reporter
• Vector type: Mammalian Expression, AAV

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: M. mazei pyrrolysyl tRNA mutant "PyOtR"
• Alt name: PyOtR
• Alt name: HTS-25
• Alt name: PytR.PyOTR
• Species: Methanosarcina mazei
• Insert Size (bp): 69
• Mutation: U25C; A3G, A4C, A5G, C6G, G63C, T65G, T66C
• Promoter: U6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsaI (destroyed during cloning)
• 3′ cloning site: BsaI (destroyed during cloning)
• 5′ sequencing primer: TTATTTACCTAGGGGTACCTCGGGCAGGAAGAGGGCCTATTTCCCATG
• 3′ sequencing primer: GATGTACTGCCAAAACCGCATCACCATGGTAATAGCGATGAC

Resource Information

• Supplemental Documents:
  • paav-itr-pytrpyotr-mcherry.gb

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry