pCMV_NESPylRS(WT)_hU6tRNAPyl
(Plasmid #223509)

• Purpose: PylRS (WT)/tRNA Pyl orthogonal pair for genetic code expansion that allows site-specific incorporation of noncanonical amino-acids into a POI using the Amber stop codon
• Depositing Lab: Edward Lemke
• Publication: Nikic et al Angew Chem Int Ed Engl. 2016 Dec 23;55(52):16172-16176. doi: 10.1002/anie.201608284. Epub 2016 Nov 2.

Backbone

• Vector backbone: pCMV
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: NES Pyrrolysyl-tRNA Synthetase
• Species: Methanosarcina mazei
• Insert Size (bp): 1392
• Entrez Gene: pylS (a.k.a. MM_RS07465, MM_1445)
• Promoter: CMV, SP6

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: n/a

Gene/Insert 2

• Gene/Insert name: tRNA Pyl
• Species: Methanosarcina mazei
• Insert Size (bp): 321
• Promoter: U6

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: unknown (unknown if destroyed)
• 3′ cloning site: unknown (unknown if destroyed)
• 5′ sequencing primer: n/a

Resource Information

• Supplemental Documents:
  • pCMV_NESPylRS(WT)_hU6tRNAPyl.gb

Terms and Licenses

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