pAS_4xMma PylT_FLAG-AbKRS-chIPYE
(Plasmid #140020)

• Purpose: Plasmid with 4xMmaPylT cassette and chimeric Mma/Mba PylRS for amber suppression and incorporation of photo-crosslinking-lysine; for transient or stable piggyBac-mediated integration
• Depositing Lab: Simon Elsaesser
• Publication: Meineke et al Cell Rep Methods. 2023 Nov 20;3(11):100626. doi: 10.1016/j.crmeth.2023.100626. Epub 2023 Nov 6.

Backbone

• Vector backbone: pAS
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

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: AbKRS-chIPYE
• Alt name: 4xh7SK-Mma PylT cassette
• Species: Methanosarcina mazei and Methanosarcina bakeri
• Insert Size (bp): 1290
• Mutation: 1-149 MbPylRS IPYE (V31I, T56P, H62Y, A100E) + 185-454MmaPylRS
• GenBank ID: Q46E77.1
• Entrez Gene: pylS (a.k.a. MSMAC_RS17105, MSMAC_3241)
• Promoter: EF1
• Tag / Fusion Protein:
  • FLAG (N terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning

Resource Information

• Supplemental Documents:
  • pas_4xmmapylt_flag-abkrs-chipye.gb
• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

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