pDONR P2R-P3-IRES_ECFP
(Plasmid
#48354)
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PurposeContributes 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.
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Depositing Labs
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 48354 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepDONRP2R-P3
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Backbone manufacturerLife Technologies/Invitrogen
- Backbone size w/o insert (bp) 2642
- Total vector size (bp) 4058
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Vector typeGateway entry vector
Growth in Bacteria
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Bacterial Resistance(s)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)Top10
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameIRES_ECFP
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SpeciesSynthetic; Aequorea victoria
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Insert Size (bp)1416
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MutationECFP 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 (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byAddgene plasmid 18673, pYIC
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
pDONR P2R-P3-IRES_ECFP was a gift from Anna Planas & Tomas Santalucia (Addgene plasmid # 48354 ; http://n2t.net/addgene:48354 ; RRID:Addgene_48354) -
For your References section:
A plasmid toolkit for cloning chimeric cDNAs encoding customized fusion proteins into any Gateway destination expression vector. Buj R, Iglesias N, Planas AM, Santalucia T. BMC Mol Biol. 2013 Aug 20;14(1):18. doi: 10.1186/1471-2199-14-18. 10.1186/1471-2199-14-18 PubMed 23957834