Stat3-C Flag pRc/CMV
(Plasmid #8722)

• Depositing Lab: Jim Darnell
• Publication: Bromberg et al Cell 1999 Aug 6;98(3):295-303.

Backbone

• Vector backbone: pRc/CMV
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5500
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Stat3-C
• Alt name: Stat3
• Species: M. musculus (mouse)
• Insert Size (bp): 2600
• Mutation: A662C and N664C. Residues changed to Cys renders the molecule constitutively active. *See comments
• GenBank ID: U06922
• Entrez Gene: Stat3 (a.k.a. 1110034C02Rik, Aprf)
• Tag / Fusion Protein:
  • Flag (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (not destroyed)
• 3′ cloning site: ApaI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: SP6

Resource Information

• Articles Citing this Plasmid:
  • 34 References

Terms and Licenses

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

Depositor Comments

Residues changed to Cys renders the molecule constitutively active. Diagnostic Restriction Digest with BamHI (3.2kb, 2.8kb, and 2.0kb). Due to a stop codon error made when tagging the Stat3 clone with FLAG (DYKDDDDK) additional residues precede (SR) and follow the tag (GPYSIVSPKC STOP). The ApaI cloning site is GGG CCC, which encodes the G and P residues coming after the FLAG tag. Therefore, if the insert is removed with a NotI and ApaI digestion, it will not have a stop codon included.

* E16K mutation found during QC