PCMT1 (NM_005389) Human Tagged ORF Clone

CAT#: RC200327

PCMT1 (Myc-DDK-tagged)-Human protein-L-isoaspartate (D-aspartate) O-methyltransferase (PCMT1)

Product images

HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY PCMT1 (Cat# RC200327, 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-PCMT1 (Cat# TA809966)(1:2000). Positive lysates LY432164 (100ug) and LC432164 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY417343) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC200327 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified PCMT1 protein (Cat# TP300327). The protein was produced from HEK293T cells transfected with PCMT1 cDNA clone (Cat# RC200327) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: PCMT1
• Synonyms: PIMT
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC200327 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGGCCTGGAAATCCGGCGGCGCCAGCCACTCGGAGCTAATCCACAATCTCCGCAAAAATGGAATCATCA
  AGACAGATAAAGTATTTGAAGTGATGCTGGCTACAGACCGCTCCCACTATGCAAAATGTAACCCATACAT
  GGATTCTCCACAATCAATAGGTTTCCAAGCAACAATCAGTGCTCCACACATGCATGCATATGCGCTAGAA
  CTTCTATTTGATCAGTTGCATGAAGGAGCTAAAGCTCTTGATGTAGGATCTGGAAGTGGAATCCTTACTG
  CATGTTTTGCACGTATGGTTGGATGTACTGGAAAAGTCATAGGAATTGATCACATTAAAGAGCTAGTAGA
  TGACTCAATAAATAATGTCAGGAAGGACGATCCAACACTTCTGTCTTCAGGGAGAGTACAGCTTGTTGTG
  GGGGATGGAAGAATGGGATATGCTGAAGAAGCCCCTTATGATGCCATTCATGTGGGAGCTGCAGCCCCTG
  TTGTACCCCAGGCGCTAATAGATCAGTTAAAGCCCGGAGGAAGATTGATATTGCCTGTTGGTCCTGCAGG
  CGGAAACCAAATGTTGGAGCAGTATGACAAGCTACAAGATGGCAGCATCAAAATGAAGCCTCTGATGGGG
  GTGATATACGTGCCTTTAACAGATAAAGAAAAGCAGTGGTCCAGGTGGAAG
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC200327 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MAWKSGGASHSELIHNLRKNGIIKTDKVFEVMLATDRSHYAKCNPYMDSPQSIGFQATISAPHMHAYALE
  LLFDQLHEGAKALDVGSGSGILTACFARMVGCTGKVIGIDHIKELVDDSINNVRKDDPTLLSSGRVQLVV
  GDGRMGYAEEAPYDAIHVGAAAPVVPQALIDQLKPGGRLILPVGPAGGNQMLEQYDKLQDGSIKMKPLMG
  VIYVPLTDKEKQWSRWK
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_005389
• ORF Size: 681 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_005389.1, NP_005380.1
• RefSeq Size: 1751 bp
• RefSeq ORF: 858 bp
• Locus ID: 5110
• UniProt ID: P22061
• Cytogenetics: 6q25.1
• Domains: PCMT
• Protein Families: Druggable Genome
• MW: 24.7 kDa
• Gene Summary: This gene encodes a member of the type II class of protein carboxyl methyltransferase enzymes. The encoded enzyme plays a role in protein repair by recognizing and converting D-aspartyl and L-isoaspartyl residues resulting from spontaneous deamidation back to the normal L-aspartyl form. The encoded protein may play a protective role in the pathogenesis of Alzheimer's disease, and single nucleotide polymorphisms in this gene have been associated with spina bifida and premature ovarian failure. Alternatively spliced transcript variants encoding multiple isoforms have been observed for this gene. [provided by RefSeq, Oct 2011]