pDONR P2R-P3-IRES_ECFP
(Plasmid #48354)

• Purpose: Contributes an IRES-ECFP cassette as the 3’-module during MultiSite Gateway cloning of a bi-cistronic mRNA shared with a two-part fusion protein encoded by the 5′- and middle modules.
• Depositing Labs: Anna Planas, Tomas Santalucia
• Publication: Buj et al BMC Mol Biol. 2013 Aug 20;14(1):18. doi: 10.1186/1471-2199-14-18.

Backbone

• Vector backbone: pDONRP2R-P3
• Backbone manufacturer: Life Technologies/Invitrogen
• Backbone size w/o insert (bp): 2642
• Total vector size (bp): 4058
• Vector type: Gateway entry vector

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Top10
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: IRES_ECFP
• Species: Synthetic; Aequorea victoria
• Insert Size (bp): 1416
• Mutation: ECFP translation is mediated by an IRES. Contains a stop codon at the end of the ECFP sequence. There is a nucleotide difference at position 1265 between our sequencing of this plasmid (A) and the reported sequence for the pYIC plasmid used as template for the PCR amplification of the insert (G). We have not confirmed whether this is related to the PCR. Nevertheless this is part of the IRES sequence and not the ECFP coding sequence.
• Promoter: none

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: M13 forward (-20)
• 3′ sequencing primer: M13 reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Addgene plasmid 18673, pYIC

Terms and Licenses

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