pcDNA3.1(+)-Histac
(Plasmid #42130)

• Depositing Lab: Philip Cole
• Publication: Dancy et al Chembiochem. 2012 Sep 24;13(14):2113-21. doi: 10.1002/cbic.201200381. Epub 2012 Sep 7.

Backbone

• Vector backbone: pcDNA3.1(+)
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 8555
• Modifications to backbone: Insert was cloned between the KpnI site and the XhoI site of the MCS (multiple cloning site).
• Vector type: Mammalian Expression, Bacterial Expression, Synthetic Biology
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Midiprep or maxiprep (larger bacterial culture than usual) for preparing DNA for mammalian transient transfection.
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Histac
• Alt name: H4AcFRETreporter
• Alt name: Venus-BRDT-H4-eCFP
• Species: Synthetic
• Insert Size (bp): 3191
• GenBank ID: NM_001242805 NM_175054
• Promoter: CMV (from pcDNA3.1(+))
• Tags / Fusion Proteins:
  • Venus (N terminal on insert)
  • eCFP (C terminal on insert)
  • Histone H4 (C terminal on insert)
  • 2 bromodomains of BRDT (N terminal on insert)
  • flexible linker

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: KpnI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: BGHrev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Minoru Yoshida

Terms and Licenses

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

Depositor Comments

First publication (with cloning info): PMID 19805290
Second publication (a review): PMID 22316554
Third publication (with applications): PMID 22961914