pSW149
(Plasmid #122248)

• Purpose: pTYB2 containing TIF4631 gene, expresses yeast eIF4G1 as an intein-chitin-binding domain fusion for purification of untagged full-length protein, with or without a fluorescent label on the C-terminus.
• Depositing Lab: Sarah Walker
• Publication: Liu et al Methods. 2019 Mar 26. pii: S1046-2023(18)30395-5. doi: 10.1016/j.ymeth.2019.03.024.

Backbone

• Vector backbone: pTYB2
• Backbone manufacturer: NEB
• Backbone size w/o insert (bp): 7474
• Total vector size (bp): 10288
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Express protein by freshly transforming in BL21-Codon Plus-DE3-RIL cells from agilent.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: TIF4631
• Alt name: eIF4G1
• Alt name: YGR162W
• Species: S. cerevisiae (budding yeast)
• Insert Size (bp): 2826
• GenBank ID: NM_001181291.3 NP_011678.3
• Entrez Gene: TIF4631 (a.k.a. YGR162W)
• Promoter: T7
• Tag / Fusion Protein:
  • Intein-Chitin-binding domain (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: TAATACGACTCACTATAGG
• 3′ sequencing primer: ACC CAT GAC CTT ATT ACC AAC CTC

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

Both plasmids pSW104 and pSW149 are being published in
Liu, X., Schuessler, P., Sahoo, A. and Walker, S.E. 2019. Reconstitution and Analyses of RNA Interactions with Eukaryotic Translation Initiation Factors and Ribosomal Preinitiation Complexes. Methods. S1046-2023: 30395-5