SHARPIN (NM_030974) Human Tagged ORF Clone

CAT#: RC222012

SHARPIN (Myc-DDK-tagged)-Human SHANK-associated RH domain interactor (SHARPIN), transcript variant 1

Product Images

Circular map for RC222012

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY SHARPIN (Cat# RC222012, 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-SHARPIN (Cat# TA811607)(1:2000). Positive lysates LY410637 (100ug) and LC410637 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY410637) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC222012 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified SHARPIN protein (Cat# TP322012). The protein was produced from HEK293T cells transfected with SHARPIN cDNA clone (Cat# RC222012) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: SHARPIN
• Synonyms: SIPL1
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC222012 representing NM_030974
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGGCGCCGCCAGCGGGCGGGGCGGCGGCGGCGGCCTCGGACTTGGGCTCCGCCGCAGTGCTCTTGGCTG
  TGCACGCCGCGGTGAGGCCGCTGGGCGCCGGGCCAGACGCCGAGGCACAGCTGCGGAGGCTGCAGCTGAG
  CGCGGACCCTGAGCGGCCTGGGCGCTTCCGGCTGGAGCTGCTGGGCGCGGGACCTGGGGCGGTTAATTTG
  GAGTGGCCCCTGGAGTCAGTTTCCTACACCATCCGAGGCCCCACCCAGCACGAGCTACAGCCTCCACCAG
  GAGGGCCTGGAACCCTCAGCCTGCACTTCCTCAACCCTCAGGAAGCTCAGCGGTGGGCAGTCCTAGTCCG
  AGGTGCCACCGTGGAAGGACAGAATGGCAGCAAGAGCAACTCACCACCAGCCTTGGGCCCAGAAGCATGC
  CCTGTCTCCCTGCCCAGTCCCCCGGAAGCCTCCACACTCAAGGGCCCTCCACCTGAGGCAGATCTTCCTA
  GGAGCCCTGGAAACTTGACGGAGAGAGAAGAGCTGGCAGGGAGCCTGGCCCGGGCTATTGCAGGTGGAGA
  CGAGAAGGGGGCAGCCCAAGTGGCAGCCGTCCTGGCCCAGCATCGTGTGGCCCTGAGTGTTCAGCTTCAG
  GAGGCCTGCTTCCCACCTGGCCCCATCAGGCTGCAGGTCACACTTGAAGACGCTGCCTCTGCCGCATCCG
  CCGCGTCCTCTGCACACGTTGCCCTGCAGGTCCACCCCCACTGCACTGTTGCAGCTCTCCAGGAGCAGGT
  GTTCTCAGAGCTCGGTTTCCCGCCAGCCGTGCAACGCTGGGTCATCGGACGGTGCCTGTGTGTGCCTGAG
  CGCAGCCTTGCCTCTTACGGGGTTCGGCAGGATGGGGACCCTGCTTTCCTCTACTTGCTGTCAGCTCCTC
  GAGAAGCCCCAGCCACAGGACCTAGCCCTCAGCACCCCCAGAAGATGGACGGGGAACTTGGACGCTTGTT
  TCCCCCATCATTGGGGCTACCCCCAGGCCCCCAGCCAGCTGCCTCCAGCCTGCCCAGTCCACTCCAGCCC
  AGCTGGTCCTGTCCTTCCTGCACCTTCATCAATGCCCCAGACCGCCCTGGCTGTGAGATGTGTAGCACCC
  AGAGGCCCTGCACTTGGGACCCCCTTGCTGCAGCTTCCACC
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC222012 representing NM_030974
  Red=Cloning site Green=Tags(s)
  
  MAPPAGGAAAAASDLGSAAVLLAVHAAVRPLGAGPDAEAQLRRLQLSADPERPGRFRLELLGAGPGAVNL
  EWPLESVSYTIRGPTQHELQPPPGGPGTLSLHFLNPQEAQRWAVLVRGATVEGQNGSKSNSPPALGPEAC
  PVSLPSPPEASTLKGPPPEADLPRSPGNLTEREELAGSLARAIAGGDEKGAAQVAAVLAQHRVALSVQLQ
  EACFPPGPIRLQVTLEDAASAASAASSAHVALQVHPHCTVAALQEQVFSELGFPPAVQRWVIGRCLCVPE
  RSLASYGVRQDGDPAFLYLLSAPREAPATGPSPQHPQKMDGELGRLFPPSLGLPPGPQPAASSLPSPLQP
  SWSCPSCTFINAPDRPGCEMCSTQRPCTWDPLAAAST
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_030974
• ORF Size: 1161 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_030974.4
• RefSeq Size: 1765 bp
• RefSeq ORF: 1164 bp
• Locus ID: 81858
• UniProt ID: Q9H0F6
• Cytogenetics: 8q24.3
• Domains: zf-RanBP
• Protein Families: Druggable Genome
• MW: 39.8 kDa
• Gene Summary: Component of the LUBAC complex which conjugates linear polyubiquitin chains in a head-to-tail manner to substrates and plays a key role in NF-kappa-B activation and regulation of inflammation. LUBAC conjugates linear polyubiquitin to IKBKG and RIPK1 and is involved in activation of the canonical NF-kappa-B and the JNK signaling pathways. Linear ubiquitination mediated by the LUBAC complex interferes with TNF-induced cell death and thereby prevents inflammation. LUBAC is recruited to the TNF-R1 signaling complex (TNF-RSC) following polyubiquitination of TNF-RSC components by BIRC2 and/or BIRC3 and to conjugate linear polyubiquitin to IKBKG and possibly other components contributing to the stability of the complex. Together with OTULIN, the LUBAC complex regulates the canonical Wnt signaling during angiogenesis.[UniProtKB/Swiss-Prot Function]