pET45b-tsr(B/X) (6X-His-Dm-Cofilin)
(Plasmid #128278)

• Purpose: Expresses Drosophila melanogaster Cofilin (Twinstar) in bacterial cell culture
• Depositing Lab: Anna Sokac
• Publication: Figard et al Cell Rep. 2019 Mar 26;26(13):3493-3501.e4. doi: 10.1016/j.celrep.2019.02.092.

Backbone

• Vector backbone: pet45b(+)
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 5260
• Total vector size (bp): 5650
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Grow with shaking at 225 rpm in Luria broth or 2X YT broth. For optimizing expression of the protein of interest, use appropriate E. coli protein expression strain such as BL21 (DE3) cells in 2X YT broth. Plasmid will be maintained at lower copy in these strains than in chemically-competent cloning strains such as E. coli Top10(TM) or DH10B(TM).
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: twinstar (tsr)
• Alt name: Dm-Cofilin
• Species: D. melanogaster (fly)
• Insert Size (bp): 452
• Mutation: Met1 changed to Pro1; Cys77 silent basepair mutation/codon switch from UGU to UGC
• GenBank ID: U24490.1 U24676.2
• Entrez Gene: tsr (a.k.a. Dmel_CG4254, 12/8, ADF, ADF/cofilin, Adf, CG4254, Cadf, D-cof, D61, Dmel\CG4254, T8, Tsr, cofilin, cofilin/ADF, cofilin/actin depolymerizing factor homolog, l(2)03101, l(2)k03101, l(2)k05633, l(2)k13409, ntf)
• Promoter: T7
• Tag / Fusion Protein:
  • 6X Histidine tag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7 (Eurofins standard primers)
• 3′ sequencing primer: T7 terminal (Eurofins standard primers)

Resource Information

• Supplemental Documents:
  • pet45-tsrBX_6X-His-Dm-Cofilin_T7term.gbk
  • pet45-tsrBX_6XHis-Dm-Cofilin_T7.gbk

Terms and Licenses

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