POLR2E (NM_002695) Human Tagged ORF Clone

CAT#: RC201266

POLR2E (Myc-DDK-tagged)-Human polymerase (RNA) II (DNA directed) polypeptide E, 25kDa (POLR2E)

Product Images

HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY POLR2E (Cat# RC201266, 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-POLR2E (Cat# TA502548). Positive lysates LY419161 (100ug) and LC419161 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY419161) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC201266 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified POLR2E protein (Cat# TP301266). The protein was produced from HEK293T cells transfected with POLR2E cDNA clone (Cat# RC201266) using MegaTran 2.0 (Cat# TT210002).

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  Anti-POLR2E mouse monoclonal antibody (TA502548) immunofluorescent staining of COS7 cells transiently transfected by pCMV6-ENTRY POLR2E (RC201266).

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  HEK293T cells transfected with either RC201266 overexpress plasmid (Red) or empty vector control plasmid (Blue) were immunostained by anti-POLR2E antibody (TA502548), and then analyzed by flow cytometry.

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Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: POLR2E
• Synonyms: hRPB25; hsRPB5; RPABC1; RPB5; XAP4
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC201266 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGGACGACGAGGAGGAGACGTACCGGCTCTGGAAAATCCGCAAGACCATCATGCAGCTGTGCCACGACC
  GTGGCTATCTGGTGACCCAGGACGAGCTTGACCAGACCCTGGAGGAGTTCAAAGCCCAATTTGGGGACAA
  GCCGAGTGAGGGGCGGCCGCGGCGCACGGACCTCACCGTGCTGGTGGCCCACAACGATGACCCCACCGAC
  CAGATGTTTGTGTTCTTTCCAGAGGAGCCCAAGGTGGGCATCAAGACCATCAAGGTGTACTGCCAGCGCA
  TGCAGGAGGAGAACATCACACGGGCTCTCATCGTGGTGCAGCAGGGCATGACACCCTCCGCCAAGCAGTC
  CCTGGTCGACATGGCCCCCAAGTACATCCTGGAGCAGTTTCTGCAGCAGGAGCTGCTCATCAACATCACG
  GAGCACGAGCTAGTCCCTGAGCACGTCGTCATGACCAAGGAGGAGGTGACAGAGCTGCTGGCCCGATATA
  AGCTCCGAGAGAACCAGCTGCCCAGGATCCAGGCGGGGGACCCTGTGGCGCGCTACTTTGGGATAAAGCG
  TGGGCAGGTGGTGAAGATCATCCGGCCCAGTGAGACGGCTGGCAGGTACATCACCTACCGGCTGGTGCAG
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC201266 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MDDEEETYRLWKIRKTIMQLCHDRGYLVTQDELDQTLEEFKAQFGDKPSEGRPRRTDLTVLVAHNDDPTD
  QMFVFFPEEPKVGIKTIKVYCQRMQEENITRALIVVQQGMTPSAKQSLVDMAPKYILEQFLQQELLINIT
  EHELVPEHVVMTKEEVTELLARYKLRENQLPRIQAGDPVARYFGIKRGQVVKIIRPSETAGRYITYRLVQ
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_002695
• ORF Size: 630 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_002695.4
• RefSeq Size: 2866 bp
• RefSeq ORF: 633 bp
• Locus ID: 5434
• UniProt ID: P19388
• Cytogenetics: 19p13.3
• Domains: RNA_pol_Rpb5_C, RNA_pol_Rpb5_N
• Protein Families: Transcription Factors
• Protein Pathways: Huntington's disease, Metabolic pathways, Purine metabolism, Pyrimidine metabolism, RNA polymerase
• MW: 24.6 kDa
• Gene Summary: This gene encodes the fifth largest subunit of RNA polymerase II, the polymerase responsible for synthesizing messenger RNA in eukaryotes. This subunit is shared by the other two DNA-directed RNA polymerases and is present in two-fold molar excess over the other polymerase subunits. An interaction between this subunit and a hepatitis virus transactivating protein has been demonstrated, suggesting that interaction between transcriptional activators and the polymerase can occur through this subunit. A pseudogene is located on chromosome 11. Three transcript variants encoding two different isoforms have been found for this gene. [provided by RefSeq, Oct 2015]