SMURF 2 (SMURF2) (NM_022739) Human Tagged ORF Clone

CAT#: RC210866

SMURF2 (Myc-DDK-tagged)-Human SMAD specific E3 ubiquitin protein ligase 2 (SMURF2)

Product Images

Circular map for RC210866

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY SMURF2 (Cat# RC210866, 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-SMURF2(Cat# TA806655). Positive lysates LY402939 (100ug) and LC402939 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY402939) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC210866 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified SMURF2 protein (Cat# TP310866). The protein was produced from HEK293T cells transfected with SMURF2 cDNA clone (Cat# RC210866) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: SMURF 2
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC210866 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGTCTAACCCCGGAGGCCGGAGGAACGGGCCCGTCAAGCTGCGCCTGACAGTACTCTGTGCAAAAAACC
  TGGTGAAAAAGGATTTTTTCCGACTTCCTGATCCATTTGCTAAGGTGGTGGTTGATGGATCTGGGCAATG
  CCATTCTACAGATACTGTGAAGAATACGCTTGATCCAAAGTGGAATCAGCATTATGACCTGTATATTGGA
  AAGTCTGATTCAGTTACGATCAGTGTATGGAATCACAAGAAGATCCATAAGAAACAAGGTGCTGGATTTC
  TCGGTTGTGTTCGTCTTCTTTCCAATGCCATCAACCGCCTCAAAGACACTGGTTATCAGAGGTTGGATTT
  ATGCAAACTCGGGCCAAATGACAATGATACAGTTAGAGGACAGATAGTAGTAAGTCTTCAGTCCAGAGAC
  CGAATAGGCACAGGAGGACAAGTTGTGGACTGCAGTCGTTTATTTGATAACGATTTACCAGACGGCTGGG
  AAGAAAGGAGAACCGCCTCTGGAAGAATCCAGTATCTAAACCATATAACAAGAACTACGCAATGGGAGCG
  CCCAACACGACCGGCATCCGAATATTCTAGCCCTGGCAGACCTCTTAGCTGCTTTGTTGATGAGAACACT
  CCAATTAGTGGAACAAATGGTGCAACATGTGGACAGTCTTCAGATCCCAGGCTGGCAGAGAGGAGAGTCA
  GGTCACAACGACATAGAAATTACATGAGCAGAACACATTTACATACTCCTCCAGACCTACCAGAAGGCTA
  TGAACAGAGGACAACGCAACAAGGCCAGGTGTATTTCTTACATACACAGACTGGTGTGAGCACATGGCAT
  GATCCAAGAGTGCCCAGGGATCTTAGCAACATCAATTGTGAAGAGCTTGGTCCATTGCCTCCTGGATGGG
  AGATCCGTAATACGGCAACAGGCAGAGTTTATTTCGTTGACCATAACAACAGAACAACACAATTTACAGA
  TCCTCGGCTGTCTGCTAACTTGCATTTAGTTTTAAATCGGCAGAACCAATTGAAAGACCAACAGCAACAG
  CAAGTGGTATCGTTATGTCCTGATGACACAGAATGCCTGACAGTCCCAAGGTACAAGCGAGACCTGGTTC
  AGAAACTAAAAATTTTGCGGCAAGAACTTTCCCAACAACAGCCTCAGGCAGGTCATTGCCGCATTGAGGT
  TTCCAGGGAAGAGATTTTTGAGGAATCATATCGACAGGTCATGAAAATGAGACCAAAAGATCTCTGGAAG
  CGATTAATGATAAAATTTCGTGGAGAAGAAGGCCTTGACTATGGAGGCGTTGCCAGGGAATGGTTGTATC
  TCTTGTCACATGAAATGTTGAATCCATACTATGGCCTCTTCCAGTATTCAAGAGATGATATTTATACATT
  GCAGATCAATCCTGATTCTGCAGTTAATCCGGAACATTTATCCTATTTCCACTTTGTTGGACGAATAATG
  GGAATGGCTGTGTTTCATGGACATTATATTGATGGTGGTTTCACATTGCCTTTTTATAAGCAATTGCTTG
  GGAAGTCAATTACCTTGGATGACATGGAGTTAGTAGATCCGGATCTTCACAACAGTTTAGTGTGGATACT
  TGAGAATGATATTACAGGTGTTTTGGACCATACCTTCTGTGTTGAACATAATGCATATGGTGAAATTATT
  CAGCATGAACTTAAACCAAATGGCAAAAGTATCCCTGTTAATGAAGAAAATAAAAAAGAATATGTCAGGC
  TCTATGTGAACTGGAGATTTTTACGAGGCATTGAGGCTCAATTCTTGGCTCTGCAGAAAGGATTTAATGA
  AGTAATTCCACAACATCTGCTGAAGACATTTGATGAGAAGGAGTTAGAGCTCATTATTTGTGGACTTGGA
  AAGATAGATGTTAATGACTGGAAGGTAAACACCCGGTTAAAACACTGTACACCAGACAGCAACATTGTCA
  AATGGTTCTGGAAAGCTGTGGAGTTTTTTGATGAAGAGCGACGAGCAAGATTGCTTCAGTTTGTGACAGG
  ATCCTCTCGAGTGCCTCTGCAGGGCTTCAAAGCATTGCAAGGTGCTGCAGGCCCGAGACTCTTTACCATA
  CACCAGATTGATGCCTGCACTAACAACCTGCCGAAAGCCCACACTTGCTTCAATCGAATAGACATTCCAC
  CCTATGAAAGCTATGAAAAGCTATATGAAAAGCTGCTAACAGCCATTGAAGAAACATGTGGATTTGCTGT
  GGAA
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC210866 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MSNPGGRRNGPVKLRLTVLCAKNLVKKDFFRLPDPFAKVVVDGSGQCHSTDTVKNTLDPKWNQHYDLYIG
  KSDSVTISVWNHKKIHKKQGAGFLGCVRLLSNAINRLKDTGYQRLDLCKLGPNDNDTVRGQIVVSLQSRD
  RIGTGGQVVDCSRLFDNDLPDGWEERRTASGRIQYLNHITRTTQWERPTRPASEYSSPGRPLSCFVDENT
  PISGTNGATCGQSSDPRLAERRVRSQRHRNYMSRTHLHTPPDLPEGYEQRTTQQGQVYFLHTQTGVSTWH
  DPRVPRDLSNINCEELGPLPPGWEIRNTATGRVYFVDHNNRTTQFTDPRLSANLHLVLNRQNQLKDQQQQ
  QVVSLCPDDTECLTVPRYKRDLVQKLKILRQELSQQQPQAGHCRIEVSREEIFEESYRQVMKMRPKDLWK
  RLMIKFRGEEGLDYGGVAREWLYLLSHEMLNPYYGLFQYSRDDIYTLQINPDSAVNPEHLSYFHFVGRIM
  GMAVFHGHYIDGGFTLPFYKQLLGKSITLDDMELVDPDLHNSLVWILENDITGVLDHTFCVEHNAYGEII
  QHELKPNGKSIPVNEENKKEYVRLYVNWRFLRGIEAQFLALQKGFNEVIPQHLLKTFDEKELELIICGLG
  KIDVNDWKVNTRLKHCTPDSNIVKWFWKAVEFFDEERRARLLQFVTGSSRVPLQGFKALQGAAGPRLFTI
  HQIDACTNNLPKAHTCFNRIDIPPYESYEKLYEKLLTAIEETCGFAVE
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_022739
• ORF Size: 2244 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_022739.4
• RefSeq Size: 3866 bp
• RefSeq ORF: 2247 bp
• Locus ID: 64750
• UniProt ID: Q9HAU4
• Cytogenetics: 17q23.3-q24.1
• Domains: C2, HECT, WW
• Protein Families: Adult stem cells, Druggable Genome, ES Cell Differentiation/IPS, Transcription Factors
• Protein Pathways: Allograft rejection, Antigen processing and presentation, Autoimmune thyroid disease, Cell adhesion molecules (CAMs), Endocytosis, Graft-versus-host disease, TGF-beta signaling pathway, Type I diabetes mellitus, Ubiquitin mediated proteolysis, Viral myocarditis
• MW: 86.2 kDa
• Gene Summary: E3 ubiquitin-protein ligase which accepts ubiquitin from an E2 ubiquitin-conjugating enzyme in the form of a thioester and then directly transfers the ubiquitin to targeted substrates. Interacts with SMAD1 and SMAD7 in order to trigger their ubiquitination and proteasome-dependent degradation. In addition, interaction with SMAD7 activates autocatalytic degradation, which is prevented by interaction with SCYE1. Forms a stable complex with the TGF-beta receptor-mediated phosphorylated SMAD2 and SMAD3. In this way, SMAD2 may recruit substrates, such as SNON, for ubiquitin-mediated degradation. Enhances the inhibitory activity of SMAD7 and reduces the transcriptional activity of SMAD2. Coexpression of SMURF2 with SMAD1 results in considerable decrease in steady-state level of SMAD1 protein and a smaller decrease of SMAD2 level.[UniProtKB/Swiss-Prot Function]