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Addgene

pcDNA3.1-IARS2-tb
(Plasmid #232340)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 232340 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pcDNA3.1 (-)
  • Backbone manufacturer
    Invitrogen (Thermo Fisher)
  • Backbone size w/o insert (bp) 5427
  • Total vector size (bp) 8445
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    IARS2
  • Alt name
    ENST00000366922.3
  • Species
    H. sapiens (human)
  • Mutation
    5 synonymous mutations in the siRNA (ACUUGCAGUCAUCCAUUAA) targeting region, in order to let the plasmid to be siRNA off-targeted.
  • GenBank ID
    NM_018060.4
  • Entrez Gene
    IARS2 (a.k.a. CAGSSS, ILERS)
  • Promoter CMV

Cloning Information

  • Cloning method Gibson Cloning
  • 5′ sequencing primer CMV-F: CGCAAATGGGCGGTAGGCGTG
  • 3′ sequencing primer BGH-R: TAGAAGGCACAGTCGAGG
    • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    Yuxin Bi (毕雨馨 in simplified Chinese), the first author of the article, constructed this plasmid during her master's degree. She graduated and become a PHD candidate in other institute for now.

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Note1:
For seamless cloning, the pcDNA3.1 (-) was single digested with Pme I and the two fragment were generated By PCR with Addgene #232337 as template and the following primers (5'->3'):
fragment 1:
3.1Pme-F: GGAGACCCAAGCTGGCTAGCGTTT
IARS2-9tb-R: tttatcaggaatcagagggtggctgcaagtacca
fragment 2:
IARS2-9tb-F: tggtacttgcagccaccctctgattcctgataaa
3.1Pme-R: ACAGTCGAGGCTGATCAGCGGTTT

Note2:
The IARS2 ORF is long, so we used additional primers (5'->3') for sanger sequencing as follow:
pc3.1-IARS2-P1: tgggaagtatgaagccaaacagt
pc3.1-IARS2-P2: gaccatggcaaaaggaacgg
pc3.1-IARS2-P3: caacagccaaaccactgagc
pc3.1-IARS2-P4: tgctcaatgctgccagaga

Note3:
The putative Kozak sequence (ENST00000366922.3:c.-6~-1) was also included

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:

    pcDNA3.1-IARS2-tb was a gift from Quanfu Ma (Addgene plasmid # 232340 ; http://n2t.net/addgene:232340 ; RRID:Addgene_232340)
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