Pyruvate Kinase (PKLR) (NM_000298) Human Tagged ORF Clone

CAT#: RC206455

PKLR (Myc-DDK-tagged)-Human pyruvate kinase, liver and RBC (PKLR), nuclear gene encoding mitochondrial protein, transcript variant 1

Product Images

Circular map for RC206455

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY PKLR (Cat# RC206455, 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-PKLR antibody (Cat# TA501413). Positive lysates LY400113 (100ug) and LC400113 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY400113) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC206455 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified PKLR protein (Cat# TP306455). The protein was produced from HEK293T cells transfected with PKLR cDNA clone (Cat# RC206455) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: Pyruvate Kinase
• Synonyms: PK1; PKL; PKRL; RPK
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC206455 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGTCGATCCAGGAGAACATATCATCCCTGCAGCTTCGGTCATGGGTCTCTAAGTCCCAAAGAGACTTAG
  CAAAGTCCATCCTGATTGGGGCTCCAGGAGGGCCAGCGGGGTATCTGCGGCGGGCCAGTGTGGCCCAACT
  GACCCAGGAGCTGGGCACTGCCTTCTTCCAGCAGCAGCAGCTGCCAGCTGCTATGGCAGACACCTTCCTG
  GAACACCTCTGCCTACTGGACATTGACTCCGAGCCCGTGGCTGCTCGCAGTACCAGCATCATTGCCACCA
  TCGGGCCAGCATCTCGCTCCGTGGAGCGCCTCAAGGAGATGATCAAGGCCGGGATGAACATTGCGCGACT
  CAACTTCTCCCACGGCTCCCACGAGTACCATGCTGAGTCCATCGCCAACGTCCGGGAGGCGGTGGAGAGC
  TTTGCAGGTTCCCCACTCAGCTACCGGCCCGTGGCCATCGCCCTGGACACCAAGGGACCGGAGATCCGCA
  CTGGGATCCTGCAGGGGGGTCCAGAGTCGGAAGTGGAGCTGGTGAAGGGCTCCCAGGTGCTGGTGACTGT
  GGACCCCGCGTTCCGGACGCGGGGGAACGCGAACACCGTGTGGGTGGACTACCCCAATATTGTCCGGGTC
  GTGCCGGTGGGGGGCCGCATCTACATTGACGACGGGCTCATCTCCCTAGTGGTCCAGAAAATCGGCCCAG
  AGGGACTGGTGACCCAAGTGGAGAACGGCGGCGTCCTGGGCAGCCGGAAGGGCGTGAACTTGCCAGGGGC
  CCAGGTGGACTTGCCCGGGCTGTCCGAGCAGGACGTCCGAGACCTGCGCTTCGGGGTGGAGCATGGGGTG
  GACATCGTCTTTGCCTCCTTTGTGCGGAAAGCCAGCGACGTGGCTGCCGTCAGGGCTGCTCTGGGTCCGG
  AAGGACACGGCATCAAGATCATCAGCAAAATTGAGAACCACGAAGGCGTGAAGAGGTTTGATGAAATCCT
  GGAGGTGAGCGACGGCATCATGGTGGCACGGGGGGACCTAGGCATCGAGATCCCAGCAGAGAAGGTTTTC
  CTGGCTCAGAAGATGATGATTGGGCGCTGCAACTTGGCGGGCAAGCCTGTTGTCTGTGCCACACAGATGC
  TGGAGAGCATGATTACCAAGCCCCGGCCAACGAGGGCAGAGACAAGCGATGTCGCCAATGCTGTGCTGGA
  TGGGGCTGACTGCATCATGCTGTCAGGGGAGACTGCCAAGGGCAACTTCCCTGTGGAAGCGGTGAAGATG
  CAGCATGCGATTGCCCGGGAGGCAGAGGCCGCAGTGTACCACCGGCAGCTGTTTGAGGAGCTACGTCGGG
  CAGCGCCACTAAGCCGTGATCCCACTGAGGTCACCGCCATTGGTGCTGTGGAGGCTGCCTTCAAGTGCTG
  TGCTGCTGCCATCATTGTGCTGACCACAACTGGCCGCTCAGCCCAGCTTCTGTCTCGGTACCGACCTCGG
  GCAGCAGTCATTGCTGTCACCCGCTCTGCCCAGGCTGCCCGCCAGGTCCACTTATGCCGAGGAGTCTTCC
  CCTTGCTTTACCGTGAACCTCCAGAAGCCATCTGGGCAGATGATGTAGATCGCCGGGTGCAATTTGGCAT
  TGAAAGTGGAAAGCTCCGTGGCTTCCTCCGTGTTGGAGACCTGGTGATTGTGGTGACAGGCTGGCGACCT
  GGCTCCGGCTACACCAACATCATGAGGGTGCTAAGCATATCC
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC206455 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MSIQENISSLQLRSWVSKSQRDLAKSILIGAPGGPAGYLRRASVAQLTQELGTAFFQQQQLPAAMADTFL
  EHLCLLDIDSEPVAARSTSIIATIGPASRSVERLKEMIKAGMNIARLNFSHGSHEYHAESIANVREAVES
  FAGSPLSYRPVAIALDTKGPEIRTGILQGGPESEVELVKGSQVLVTVDPAFRTRGNANTVWVDYPNIVRV
  VPVGGRIYIDDGLISLVVQKIGPEGLVTQVENGGVLGSRKGVNLPGAQVDLPGLSEQDVRDLRFGVEHGV
  DIVFASFVRKASDVAAVRAALGPEGHGIKIISKIENHEGVKRFDEILEVSDGIMVARGDLGIEIPAEKVF
  LAQKMMIGRCNLAGKPVVCATQMLESMITKPRPTRAETSDVANAVLDGADCIMLSGETAKGNFPVEAVKM
  QHAIAREAEAAVYHRQLFEELRRAAPLSRDPTEVTAIGAVEAAFKCCAAAIIVLTTTGRSAQLLSRYRPR
  AAVIAVTRSAQAARQVHLCRGVFPLLYREPPEAIWADDVDRRVQFGIESGKLRGFLRVGDLVIVVTGWRP
  GSGYTNIMRVLSIS
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_000298
• ORF Size: 1722 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_000298.2
• RefSeq Size: 3053 bp
• RefSeq ORF: 1725 bp
• Locus ID: 5313
• UniProt ID: P30613
• Cytogenetics: 1q22
• Domains: PK
• Protein Families: Druggable Genome
• Protein Pathways: Glycolysis / Gluconeogenesis, Insulin signaling pathway, Maturity onset diabetes of the young, Metabolic pathways, Purine metabolism, Pyruvate metabolism, Type II diabetes mellitus
• MW: 61.8 kDa
• Gene Summary: The protein encoded by this gene is a pyruvate kinase that catalyzes the transphosphorylation of phohsphoenolpyruvate into pyruvate and ATP, which is the rate-limiting step of glycolysis. Defects in this enzyme, due to gene mutations or genetic variations, are the common cause of chronic hereditary nonspherocytic hemolytic anemia (CNSHA or HNSHA). Multiple transcript variants encoding different isoforms have been found for this gene. [provided by RefSeq, Jul 2008]