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pMIR-Report-SPRED1-3'UTR (mut - miR-21 site 2)
(Plasmid #62581)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 62581 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pMIR-Report
  • Backbone manufacturer
    Ambion
  • Backbone size w/o insert (bp) 6470
  • Total vector size (bp) 7214
  • Modifications to backbone
    The downstream miR-21 binding site (site 2) was altered using PCR mutagenesis (TAAGCTA --> TAGATCA) To construct a wild-type SPRED1 reporter containing the miR-21 binding sites, a 744-bp fragment of the SPRED1 3′ UTR containing two putative miR-21 binding sites was amplified by PCR from MDA-MB-231 cDNA using the oligonucleotides 5′-cccacgcgtTGAAAAACTGTTTAACTCATGT-3′ and 5′-cccacgcgtTGAAAAACCTGTAAATAAGCAC-3′. (The SPRED1 sequence is indicated in uppercase.) Following MluI digestion, the product was cloned into the same site of the pMIR-REPORT vector.
  • Vector type
    Luciferase

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    XL1 Blue
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    SPRED1
  • Alt name
    Sprouty Related EVH1 domain containing 1
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    744
  • Mutation
    miR-21 binding site 2 mutant (TAAGCTA --> TAGATCA)
  • Entrez Gene
    SPRED1 (a.k.a. LGSS, NFLS, PPP1R147, hSpred1, spred-1)

Cloning Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

There are a few mismatches outside of the mutated seed sequence; these mismatches should not affect plasmid function.

How to cite this plasmid ( Back to top)

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.

  • For your Materials & Methods section:

    pMIR-Report-SPRED1-3'UTR (mut - miR-21 site 2) was a gift from Michael Ruppert (Addgene plasmid # 62581 ; http://n2t.net/addgene:62581 ; RRID:Addgene_62581)

  • For your References section:

    MicroRNAs 206 and 21 cooperate to promote RAS-extracellular signal-regulated kinase signaling by suppressing the translation of RASA1 and SPRED1. Sharma SB, Lin CC, Farrugia MK, McLaughlin SL, Ellis EJ, Brundage KM, Salkeni MA, Ruppert JM. Mol Cell Biol. 2014 Nov 15;34(22):4143-64. doi: 10.1128/MCB.00480-14. Epub 2014 Sep 8. 10.1128/MCB.00480-14 PubMed 25202123
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