HLCS (NM_000411) Human Tagged ORF Clone

CAT#: RC209197

HLCS (Myc-DDK-tagged)-Human holocarboxylase synthetase (biotin-(proprionyl-CoA-carboxylase (ATP-hydrolysing)) ligase) (HLCS)

Product Images

Circular map for RC209197

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY HLCS (Cat# RC209197, 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-HLCS(Cat# TA504422). Positive lysates LY400145 (100ug) and LC400145 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY400145) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC209197 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified HLCS protein (Cat# TP309197). The protein was produced from HEK293T cells transfected with HLCS cDNA clone (Cat# RC209197) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: HLCS
• Synonyms: HCS
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC209197 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGGAAGATAGACTCCACATGGATAATGGACTGGTACCCCAAAAGATTGTGTCGGTGCACTTGCAGGACT
  CCACTCTGAAGGAAGTTAAGGATCAGGTCTCAAACAAGCAAGCCCAGATCCTAGAGCCGAAGCCTGAACC
  TTCTCTTGAGATTAAGCCTGAGCAGGACGGTATGGAGCATGTTGGCAGAGATGACCCAAAGGCTCTTGGT
  GAAGAACCCAAACAAAGGAGAGGCAGTGCCTCTGGGAGTGAGCCTGCTGGGGACAGTGACAGGGGAGGGG
  GCCCCGTTGAGCATTATCACCTCCATCTGTCTAGTTGCCACGAGTGTCTGGAACTTGAGAACAGCACCAT
  TGAGTCAGTCAAGTTTGCGTCTGCCGAGAACATTCCAGACCTTCCCTACGATTATAGCAGCAGTTTGGAG
  AGTGTTGCTGATGAGACCTCCCCCGAAAGAGAAGGGAGGAGAGTCAACCTCACGGGAAAGGCACCCAACA
  TCCTCCTCTATGTGGGCTCCGACTCCCAGGAAGCCCTCGGCCGGTTCCACGAGGTCCGGTCTGTGCTGGC
  CGACTGTGTGGACATTGACAGTTATATTCTCTACCACCTGCTGGAGGACAGTGCTCTCAGAGACCCGTGG
  ACGGACAACTGTCTGCTGTTGGTCATTGCTACCAGGGAGTCCATTCCCGAAGACCTGTACCAGAAGTTCA
  TGGCCTATCTTTCTCAGGGAGGGAAGGTGTTGGGCCTGTCTTCATCCTTCACCTTTGGTGGCTTTCAGGT
  GACAAGCAAGGGTGCACTGCACAAGACAGTCCAGAACTTGGTTTTCTCCAAGGCTGACCAGAGTGAGGTG
  AAGCTCAGCGTCTTGAGCAGTGGCTGCAGGTACCAGGAAGGCCCCGTCCGGCTCAGCCCCGGCAGGCTCC
  AGGGCCACCTGGAGAATGAGGACAAGGACAGGATGATTGTGCATGTGCCTTTTGGAACTCGCGGGGGAGA
  AGCTGTTCTTTGCCAGGTGCACTTAGAACTACCTCCCAGCTCCAACATAGTGCAAACTCCAGAAGATTTT
  AACTTGCTCAAGTCAAGCAATTTTAGAAGATACGAAGTCCTTAGAGAGATTCTGACAACCCTTGGCCTCA
  GCTGTGACATGAAACAAGTTCCTGCCTTAACTCCTCTTTACTTGCTGTCAGCTGCGGAGGAAATCAGGGA
  TCCTCTTATGCAGTGGCTTGGGAAACATGTGGACTCCGAGGGAGAAATAAAATCCGGCCAGCTCTCTCTT
  AGATTTGTTTCATCCTACGTGTCTGAAGTAGAAATAACCCCATCTTGTATACCTGTGGTGACCAACATGG
  AGGCCTTCTCATCAGAACATTTCAACTTAGAGATCTATCGCCAAAATCTGCAGACCAAGCAGTTGGGGAA
  AGTAATTTTGTTTGCCGAAGTGACCCCCACAACGATGCGTCTCCTGGATGGGCTGATGTTTCAGACACCG
  CAGGAAATGGGCTTAATAGTGATCGCGGCCCGGCAGACCGAGGGCAAAGGACGGGGAGGGAATGTGTGGC
  TGAGCCCTGTGGGATGTGCTCTTTCTACTCTGCTCATCTCCATTCCACTGAGATCCCAGCTGGGACAGAG
  GATCCCGTTTGTCCAGCATCTGATGTCCGTGGCTGTCGTGGAAGCAGTGAGGTCCATTCCCGAGTATCAG
  GATATCAACTTACGAGTGAAGTGGCCCAACGATATTTATTACAGTGACCTCATGAAGATCGGCGGAGTTC
  TGGTTAACTCAACACTCATGGGAGAAACATTTTATATACTTATTGGCTGTGGATTTAATGTGACTAACAG
  TAACCCTACCATCTGCATCAACGACCTCATCACAGAATACAATAAACAACACAAGGCAGAACTGAAGCCC
  TTAAGAGCCGATTATCTCATCGCCAGAGTCGTGACTGTGCTGGAGAAACTGATCAAAGAGTTTCAGGACA
  AAGGGCCCAACAGCGTCCTTCCCCTTTATTACCGATACTGGGTCCACAGTGGTCAGCAAGTCCATCTGGG
  CAGCGCAGAGGGACCAAAGGTGTCCATCGTTGGCCTGGACGATTCTGGCTTCCTCCAGGTTCACCAGGAG
  GGCGGCGAGGTTGTGACTGTGCACCCGGACGGCAACTCCTTCGACATGCTGAGAAACCTCATCCTCCCCA
  AACGGCGG
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC209197 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MEDRLHMDNGLVPQKIVSVHLQDSTLKEVKDQVSNKQAQILEPKPEPSLEIKPEQDGMEHVGRDDPKALG
  EEPKQRRGSASGSEPAGDSDRGGGPVEHYHLHLSSCHECLELENSTIESVKFASAENIPDLPYDYSSSLE
  SVADETSPEREGRRVNLTGKAPNILLYVGSDSQEALGRFHEVRSVLADCVDIDSYILYHLLEDSALRDPW
  TDNCLLLVIATRESIPEDLYQKFMAYLSQGGKVLGLSSSFTFGGFQVTSKGALHKTVQNLVFSKADQSEV
  KLSVLSSGCRYQEGPVRLSPGRLQGHLENEDKDRMIVHVPFGTRGGEAVLCQVHLELPPSSNIVQTPEDF
  NLLKSSNFRRYEVLREILTTLGLSCDMKQVPALTPLYLLSAAEEIRDPLMQWLGKHVDSEGEIKSGQLSL
  RFVSSYVSEVEITPSCIPVVTNMEAFSSEHFNLEIYRQNLQTKQLGKVILFAEVTPTTMRLLDGLMFQTP
  QEMGLIVIAARQTEGKGRGGNVWLSPVGCALSTLLISIPLRSQLGQRIPFVQHLMSVAVVEAVRSIPEYQ
  DINLRVKWPNDIYYSDLMKIGGVLVNSTLMGETFYILIGCGFNVTNSNPTICINDLITEYNKQHKAELKP
  LRADYLIARVVTVLEKLIKEFQDKGPNSVLPLYYRYWVHSGQQVHLGSAEGPKVSIVGLDDSGFLQVHQE
  GGEVVTVHPDGNSFDMLRNLILPKRR
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_000411
• ORF Size: 2178 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_000411.8
• RefSeq Size: 6019 bp
• RefSeq ORF: 2181 bp
• Locus ID: 3141
• UniProt ID: P50747
• Cytogenetics: 21q22.13
• Domains: BPL_C, BPL_LipA_LipB
• Protein Pathways: Biotin metabolism, Metabolic pathways
• MW: 80.8 kDa
• Gene Summary: This gene encodes an enzyme that catalyzes the binding of biotin to carboxylases and histones. The protein plays an important role in gluconeogenesis, fatty acid synthesis and branched chain amino acid catabolism. Defects in this gene are the cause of holocarboxylase synthetase deficiency. Multiple alternatively spliced variants, encoding the same protein, have been identified.[provided by RefSeq, Jun 2011]