DBT (NM_001918) Human Tagged ORF Clone

CAT#: RC201998

DBT (Myc-DDK-tagged)-Human dihydrolipoamide branched chain transacylase E2 (DBT), nuclear gene encoding mitochondrial protein

Product Images

Circular map for RC201998

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY DBT (Cat# RC201998, Right lane) cDNA for 48 hrs and lysed. Equivalent amounts of cell lysates (5 ug per lane) were separated by SDS-PAGE and immunoblotted with anti-DBT(Cat# TA504748). Positive lysates LY419653 (100ug) and LC419653 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY419653) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC201998 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified DBT protein (Cat# TP301998). The protein was produced from HEK293T cells transfected with DBT cDNA clone (Cat# RC201998) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: DBT
• Synonyms: BCATE2; BCKAD-E2; BCKADE2; BCKDH-E2; BCOADC-E2; E2; E2B
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC201998 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGGCTGCAGTCCGTATGCTGAGAACCTGGAGCAGGAATGCGGGGAAGCTGATTTGTGTTCGCTATTTTC
  AAACATGTGGTAATGTTCATGTTTTGAAGCCAAATTATGTGTGTTTCTTTGGTTATCCTTCATTCAAGTA
  TAGTCATCCACATCACTTCCTGAAAACAACTGCTGCTCTCCGTGGACAGGTTGTTCAGTTCAAGCTCTCA
  GACATTGGAGAAGGGATTAGAGAAGTAACTGTTAAAGAATGGTATGTAAAAGAAGGAGATACAGTGTCTC
  AGTTTGATAGCATCTGTGAAGTTCAAAGTGATAAAGCTTCTGTTACCATCACTAGTCGTTATGATGGAGT
  CATTAAAAAACTCTATTATAATCTAGACGATATTGCCTATGTGGGGAAGCCATTAGTAGACATAGAAACG
  GAAGCTTTAAAAGATTCAGAAGAAGATGTTGTTGAAACTCCTGCAGTGTCTCATGATGAACATACACACC
  AAGAGATAAAGGGCCGAAAAACACTGGCAACTCCTGCAGTTCGCCGTCTGGCAATGGAAAACAATATTAA
  GCTGAGTGAAGTTGTTGGCTCAGGAAAAGATGGCAGAATACTTAAAGAAGATATCCTCAACTATTTGGAA
  AAGCAGACAGGAGCTATATTGCCTCCTTCACCCAAAGTTGAAATTATGCCACCTCCACCAAAGCCAAAAG
  ACATGACTGTTCCTATACTAGTATCAAAACCTCCGGTATTCACAGGCAAAGACAAAACAGAACCCATAAA
  AGGCTTTCAAAAAGCAATGGTCAAGACTATGTCTGCAGCCCTGAAGATACCTCATTTTGGTTATTGTGAT
  GAGATTGACCTTACTGAACTGGTTAAGCTCCGAGAAGAATTAAAACCCATTGCATTTGCTCGTGGAATTA
  AACTCTCCTTTATGCCTTTCTTCTTAAAGGCTGCTTCCTTGGGATTACTACAGTTTCCTATCCTTAACGC
  TTCTGTGGATGAAAACTGCCAGAATATAACATATAAGGCTTCTCATAACATTGGGATAGCAATGGATACT
  GAGCAGGGTTTGATTGTCCCTAATGTGAAAAATGTTCAGATCTGCTCTATATTTGACATCGCCACTGAAC
  TGAACCGCCTCCAGAAATTGGGCTCTGTGGGTCAGCTCAGCACCACTGATCTTACAGGAGGAACATTTAC
  TCTTTCCAACATTGGATCAATTGGTGGTACCTTTGCCAAACCAGTGATAATGCCACCTGAAGTAGCCATT
  GGGGCCCTTGGATCAATTAAGGCCATTCCCCGATTTAACCAGAAAGGAGAAGTATATAAGGCACAGATAA
  TGAATGTGAGCTGGTCAGCTGATCACAGAGTTATTGATGGTGCTACAATGTCACGCTTCTCCAATTTGTG
  GAAATCCTATTTAGAAAACCCAGCTTTTATGCTACTAGATCTGAAA
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC201998 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MAAVRMLRTWSRNAGKLICVRYFQTCGNVHVLKPNYVCFFGYPSFKYSHPHHFLKTTAALRGQVVQFKLS
  DIGEGIREVTVKEWYVKEGDTVSQFDSICEVQSDKASVTITSRYDGVIKKLYYNLDDIAYVGKPLVDIET
  EALKDSEEDVVETPAVSHDEHTHQEIKGRKTLATPAVRRLAMENNIKLSEVVGSGKDGRILKEDILNYLE
  KQTGAILPPSPKVEIMPPPPKPKDMTVPILVSKPPVFTGKDKTEPIKGFQKAMVKTMSAALKIPHFGYCD
  EIDLTELVKLREELKPIAFARGIKLSFMPFFLKAASLGLLQFPILNASVDENCQNITYKASHNIGIAMDT
  EQGLIVPNVKNVQICSIFDIATELNRLQKLGSVGQLSTTDLTGGTFTLSNIGSIGGTFAKPVIMPPEVAI
  GALGSIKAIPRFNQKGEVYKAQIMNVSWSADHRVIDGATMSRFSNLWKSYLENPAFMLLDLK
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_001918
• ORF Size: 1446 bp
• OTI Disclaimer: The molecular sequence of this clone aligns with the gene accession number as a point of reference only. However, individual transcript sequences of the same gene can differ through naturally occurring variations (e.g. polymorphisms), each with its own valid existence. This clone is substantially in agreement with the reference, but a complete review of all prevailing variants is recommended prior to use. More info
• OTI Annotation: This clone was engineered to express the complete ORF with an expression tag. Expression varies depending on the nature of the gene.
• Product Components: The ORF clone is ion-exchange column purified and shipped in a 2D barcoded Matrix tube containing 10ug of transfection-ready, dried plasmid DNA (reconstitute with 100 ul of water).
• Reconstitution: 1. Centrifuge at 5,000xg for 5min.
  2. Carefully open the tube and add 100ul of sterile water to dissolve the DNA.
  3. Close the tube and incubate for 10 minutes at room temperature.
  4. Briefly vortex the tube and then do a quick spin (less than 5000xg) to concentrate the liquid at the bottom.
  5. Store the suspended plasmid at -20°C. The DNA is stable for at least one year from date of shipping when stored at -20°C.

Reference Data

• RefSeq: NM_001918.4
• RefSeq Size: 10831 bp
• RefSeq ORF: 1449 bp
• Locus ID: 1629
• UniProt ID: P11182
• Cytogenetics: 1p21.2
• Domains: biotin_lipoyl, 2-oxoacid_dh, e3_binding
• Protein Families: Druggable Genome
• Protein Pathways: Metabolic pathways, Valine, leucine and isoleucine degradation
• MW: 53.5 kDa
• Gene Summary: The branched-chain alpha-keto acid dehydrogenase complex (BCKD) is an inner-mitochondrial enzyme complex involved in the breakdown of the branched-chain amino acids isoleucine, leucine, and valine. The BCKD complex is thought to be composed of a core of 24 transacylase (E2) subunits, and associated decarboxylase (E1), dehydrogenase (E3), and regulatory subunits. This gene encodes the transacylase (E2) subunit. Mutations in this gene result in maple syrup urine disease, type 2. Alternatively spliced transcript variants have been described, but their biological validity has not been determined. [provided by RefSeq, Jul 2008]