pAJE3-E7
(Plasmid #214359)

• Purpose: Expresses the 2nd-generation "E7" Tet2 Methanocaldococcus jannaschii (Mj) aminoacyl tRNA synthetase/tRNA pair for encoding Tet2 noncanonical amino acids into TAG sites of proteins
• Depositing Lab: Ryan Mehl
• Publication: Eddins et al Bioconjug Chem. 2023 Dec 20;34(12):2243-2254. doi: 10.1021/acs.bioconjchem.3c00380. Epub 2023 Dec 4.

Backbone

• Vector backbone: pAJE3
• Backbone size w/o insert (bp): 5927
• Total vector size (bp): 6848
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: 2nd generation Tet2 "E7" aminoacyl tRNA synthetase
• Alt name: E7 aaRS
• Species: M. jannaschii
• Insert Size (bp): 921
• Promoter: lpp

Cloning Information for Gene/Insert 1

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

Gene/Insert 2

• Gene/Insert name: Lpp promoted M. jannaschii tRNA
• Alt name: Mj tRNA 1.0
• Species: M. jannaschii
• Insert Size (bp): 142
• Promoter: lpp

Cloning Information for Gene/Insert 2

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

Gene/Insert 3

• Gene/Insert name: Aminoglycoside-3''-adenyltransferase (Spectinomycin/streptomycin Resistance Gene)
• Insert Size (bp): 792
• Promoter: AmpR

Cloning Information for Gene/Insert 3

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

Gene/Insert 4

• Gene/Insert name: Orthogonal "D4II" ColE1 origin
• Alt name: D4II
• Species: Synthetic
• Insert Size (bp): 588

Resource Information

• Supplemental Documents:
  • pAJE3_E7_Tet2.0Et-edited-with-mutations.gbk

Terms and Licenses

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