pBT270_(pCA-G-intron(Neo)-tTA2-iiTRE-tdT3Mycii)
(Plasmid #36881)

• Depositing Lab: Liqun Luo
• Publication: Tasic et al PLoS One. 2012;7(3):e33332. Epub 2012 Mar 27.

Backbone

• Vector backbone: pBluescript SK(+)
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 3000
• Total vector size (bp): 11396
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): Stbl2
• Growth instructions: If plasmid yields are poor in DH5alpha, we recommend Stbl2 cells (Invitrogen) grown at 30C.
• Copy number: High Copy

Gene/Insert 1

• Gene/Insert name: GFP
• Alt name: mut4EGFP
• Mutation: deleted nucleotides after nucleotide 274
• Promoter: CAG (chicken beta actin promoter and CMV enhancer)

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: see sequence
• 3′ sequencing primer: see sequence

Gene/Insert 2

• Gene/Insert name: tTA2
• Alt name: tetracycline transactivator
• Mutation: removed start codon
• Promoter: CAG (chicken beta actin promoter and CMV enhancer)

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: see sequence
• 3′ sequencing primer: see sequence

Gene/Insert 3

• Gene/Insert name: beta-globin intron
• Mutation: insertion of FRT, lox5171, lox2272, FRT, loxP-Neo-loxP
• Promoter: CAG (chicken beta actin promoter and CMV enhancer)

Cloning Information for Gene/Insert 3

• Cloning method: Unknown
• 5′ sequencing primer: see sequence
• 3′ sequencing primer: see sequence

Gene/Insert 4

• Gene/Insert name: Neo
• Alt name: neomycin phosphotransferase
• Alt name: neomycin resistance gene
• Promoter: SV40

Cloning Information for Gene/Insert 4

• Cloning method: Unknown
• 5′ sequencing primer: see sequence
• 3′ sequencing primer: see sequence

Gene/Insert 5

• Gene/Insert name: tdT-3Myc
• Alt name: tdTomato
• Promoter: TRE (tetO)
• Tag / Fusion Protein:
  • 3 Myc tags (C terminal on insert)

Cloning Information for Gene/Insert 5

• Cloning method: Unknown
• 5′ sequencing primer: see sequence
• 3′ sequencing primer: see sequence

Gene/Insert 6

• Gene/Insert name: insulator
• Alt name: chicken beta globin HS4 insulator core (~250 bp)
• Promoter: none

Cloning Information for Gene/Insert 6

• Cloning method: Unknown
• 5′ sequencing primer: see sequence
• 3′ sequencing primer: T3

Resource Information

• Supplemental Documents:
  • pBT270_GenBank_file
• A portion of this plasmid was derived from a plasmid made by: W. Hillen, R. Tsien and G. Felsenfeld

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry