TPMT (NM_000367) Human Tagged ORF Clone

CAT#: RC203309

TPMT (Myc-DDK-tagged)-Human thiopurine S-methyltransferase (TPMT)

Product Images

Circular map for RC203309

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY TPMT (Cat# RC203309, 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-TPMT(Cat# TA500966). Positive lysates LY400131 (100ug) and LC400131 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY400131) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC203309 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified TPMT protein (Cat# TP303309). The protein was produced from HEK293T cells transfected with TPMT cDNA clone (Cat# RC203309) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

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

  >RC203309 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGGATGGTACAAGAACTTCACTTGACATTGAAGAGTACTCGGATACTGAGGTACAGAAAAACCAAGTAC
  TAACTCTGGAAGAATGGCAAGACAAGTGGGTGAACGGCAAGACTGCTTTTCATCAGGAACAAGGACATCA
  GCTATTAAAGAAGCATTTAGATACTTTCCTTAAAGGCAAGAGTGGACTGAGGGTATTTTTTCCTCTTTGC
  GGAAAAGCGGTTGAGATGAAATGGTTTGCAGACCGGGGACACAGTGTAGTTGGTGTGGAAATCAGTGAAC
  TTGGGATACAAGAATTTTTTACAGAGCAGAATCTTTCTTACTCAGAAGAACCAATCACCGAAATTCCTGG
  AACCAAAGTATTTAAGAGTTCTTCGGGGAACATTTCATTGTACTGTTGCAGTATTTTTGATCTTCCCAGG
  ACAAATATTGGCAAATTTGACATGATTTGGGATAGAGGAGCATTAGTTGCCATTAATCCAGGTGATCGCA
  AATGCTATGCAGATACAATGTTTTCCCTCCTGGGAAAGAAGTTTCAGTATCTCCTGTGTGTTCTTTCTTA
  TGATCCAACTAAACATCCAGGTCCACCATTTTATGTTCCACATGCTGAAATTGAAAGGTTGTTTGGTAAA
  ATATGCAATATACGTTGTCTTGAGAAGGTTGATGCTTTTGAAGAACGACATAAAAGTTGGGGAATTGACT
  GTCTTTTTGAAAAGTTATATCTACTTACAGAAAAG
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC203309 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MDGTRTSLDIEEYSDTEVQKNQVLTLEEWQDKWVNGKTAFHQEQGHQLLKKHLDTFLKGKSGLRVFFPLC
  GKAVEMKWFADRGHSVVGVEISELGIQEFFTEQNLSYSEEPITEIPGTKVFKSSSGNISLYCCSIFDLPR
  TNIGKFDMIWDRGALVAINPGDRKCYADTMFSLLGKKFQYLLCVLSYDPTKHPGPPFYVPHAEIERLFGK
  ICNIRCLEKVDAFEERHKSWGIDCLFEKLYLLTEK
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_000367
• ORF Size: 735 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_000367.4
• RefSeq Size: 3281 bp
• RefSeq ORF: 738 bp
• Locus ID: 7172
• UniProt ID: P51580
• Cytogenetics: 6p22.3
• Protein Families: Druggable Genome
• Protein Pathways: Drug metabolism - other enzymes
• MW: 28.2 kDa
• Gene Summary: This gene encodes the enzyme that metabolizes thiopurine drugs via S-adenosyl-L-methionine as the S-methyl donor and S-adenosyl-L-homocysteine as a byproduct. Thiopurine drugs such as 6-mercaptopurine are used as chemotherapeutic agents. Genetic polymorphisms that affect this enzymatic activity are correlated with variations in sensitivity and toxicity to such drugs within individuals, causing thiopurine S-methyltransferase deficiency. Related pseudogenes have been identified on chromosomes 3, 18 and X. [provided by RefSeq, Aug 2014]