H6hSDHA
(Plasmid #113045)

• Purpose: Hetero expression of His-tagged human SDHA
• Depositing Lab: Edward A. Berry
• Publication: Zafreen et al Arch Biochem Biophys. 2016 Aug 15;604:47-56. doi: 10.1016/j.abb.2016.06.006. Epub 2016 Jun 11.

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, PPGL5, 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

Terms and Licenses

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