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H6hSDHA
(Plasmid #113045)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 113045 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET_Duet-1
  • Modifications to backbone
    The expression vector for N-terminally His6-tagged mature human SDHA (H6hSDHA) was constructed by placing the nucleotides coding for the mature sequence (starting with residues 43ASAK, through the stop codon) into the first multicloning site of the pETDuet™-1 vector between the BamHI and EcoRI restriction sites. This results in prepending 14 residues MGSSH6SQDP from coding sequence in the vector. It is expressed in BL-21 cells together with pRARE plasmid to support rare codons. In the absence of hSDH5 it is flavinylated little if at all, and mostly goes into inclusion bodies.
  • Vector type
    Bacterial Expression

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    BL-21
  • Growth instructions
    Cells may also contain pRARE. To confirm the presence of this plasmid select with chloramphenicol. For protein expression (induced with IPTG) grow at 30C but for growing cells without IPTG grow at 37C.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    H6hSDHA
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    1908
  • Mutation
    Insert begins at amino acid 43
  • GenBank ID
    LT742803.1
  • Entrez Gene
    SDHA (a.k.a. CMD1GG, FP, MC2DN1, NDAXOA, PGL5, SDH1, SDH2, SDHF)
  • Tag / Fusion Protein
    • His6 (MGSSHHHHHHSQDP-) (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site unknown (unknown if destroyed)
  • 3′ cloning site unknown (unknown if destroyed)
  • 5′ sequencing primer unknown
  • (Common Sequencing Primers)

Terms and Licenses

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

    H6hSDHA was a gift from Edward A. Berry (Addgene plasmid # 113045 ; http://n2t.net/addgene:113045 ; RRID:Addgene_113045)
  • For your References section:

    In-vitro, SDH5-dependent flavinylation of immobilized human respiratory complex II flavoprotein. Zafreen L, Walker-Kopp N, Huang LS, Berry E. Arch Biochem Biophys. 2016 Aug 15;604:47-56. doi: 10.1016/j.abb.2016.06.006. Epub 2016 Jun 11. 10.1016/j.abb.2016.06.006 PubMed 27296776