pGEMT-PT2A-iRFP670-Tdtomato-GFP
(Plasmid #111817)

• Purpose: Tricistronic construct: Co-express three fluorescent proteins by P2A and then T2A
• Depositing Lab: Li Qian
• Publication: Liu et al Sci Rep. 2017 May 19;7(1):2193. doi: 10.1038/s41598-017-02460-2.

Backbone

• Vector backbone: pGEMT-easy
• Backbone size w/o insert (bp): 3000
• Total vector size (bp): 6200
• Vector type: cloning intermediate

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): HB101
• Copy number: Low Copy

Gene/Insert 1

• Gene/Insert name: iRFP670
• Insert Size (bp): 900

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: NheI (destroyed during cloning)
• 5′ sequencing primer: M13 F/T7
• 3′ sequencing primer: P2A-r

Gene/Insert 2

• Gene/Insert name: Tdtomato
• Insert Size (bp): 1400

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: SpeI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: P2A-f
• 3′ sequencing primer: T2A-r

Gene/Insert 3

• Gene/Insert name: GFP
• Insert Size (bp): 700

Cloning Information for Gene/Insert 3

• Cloning method: Restriction Enzyme
• 5′ cloning site: XhoI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: T2A-f
• 3′ sequencing primer: M13R/SP6

Resource Information

• Addgene Notes:
  • Addgene Diagnostic Digest

Terms and Licenses

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