MDA5 (IFIH1) (NM_022168) Human Tagged ORF Clone

CAT#: RC215661

IFIH1 (Myc-DDK-tagged)-Human interferon induced with helicase C domain 1 (IFIH1)

Product Images

Circular map for RC215661

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY IFIH1 (Cat# RC215661, 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-IFIH1(Cat# TA803667). Positive lysates LY411723 (100ug) and LC411723 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY411723) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC215661 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified IFIH1 protein (Cat# TP315661). The protein was produced from HEK293T cells transfected with IFIH1 cDNA clone (Cat# RC215661) using MegaTran 2.0 (Cat# TT210002).

Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: MDA5
• Synonyms: AGS7; Hlcd; IDDM19; MDA-5; MDA5; RLR-2; SGMRT1
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC215661 representing NM_022168
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGTCGAATGGGTATTCCACAGACGAGAATTTCCGCTATCTCATCTCGTGCTTCAGGGCCAGGGTGAAAA
  TGTACATCCAGGTGGAGCCTGTGCTGGACTACCTGACCTTTCTGCCTGCAGAGGTGAAGGAGCAGATTCA
  GAGGACAGTCGCCACCTCCGGGAACATGCAGGCAGTTGAACTGCTGCTGAGCACCTTGGAGAAGGGAGTC
  TGGCACCTTGGTTGGACTCGGGAATTCGTGGAGGCCCTCCGGAGAACCGGCAGCCCTCTGGCCGCCCGCT
  ACATGAACCCTGAGCTCACGGACTTGCCCTCTCCATCGTTTGAGAACGCTCATGATGAATATCTCCAACT
  GCTGAACCTCCTTCAGCCCACTCTGGTGGACAAGCTTCTAGTTAGAGACGTCTTGGATAAGTGCATGGAG
  GAGGAACTGTTGACAATTGAAGACAGAAACCGGATTGCTGCTGCAGAAAACAATGGAAATGAATCAGGTG
  TAAGAGAGCTACTAAAAAGGATTGTGCAGAAAGAAAACTGGTTCTCTGCATTTCTGAATGTTCTTCGTCA
  AACAGGAAACAATGAACTTGTCCAAGAGTTAACAGGCTCTGATTGCTCAGAAAGCAATGCAGAGATTGAG
  AATTTATCACAAGTTGATGGTCCTCAAGTGGAAGAGCAACTTCTTTCAACCACAGTTCAGCCAAATCTGG
  AGAAGGAGGTCTGGGGCATGGAGAATAACTCATCAGAATCATCTTTTGCAGATTCTTCTGTAGTTTCAGA
  ATCAGACACAAGTTTGGCAGAAGGAAGTGTCAGCTGCTTAGATGAAAGTCTTGGACATAACAGCAACATG
  GGCAGTGATTCAGGCACCATGGGAAGTGATTCAGATGAAGAGAATGTGGCAGCAAGAGCATCCCCGGAGC
  CAGAACTCCAGCTCAGGCCTTACCAAATGGAAGTTGCCCAGCCAGCCTTGGAAGGGAAGAATATCATCAT
  CTGCCTCCCTACAGGGAGTGGAAAAACCAGAGTGGCTGTTTACATTGCCAAGGATCACTTAGACAAGAAG
  AAAAAAGCATCTGAGCCTGGAAAAGTTATAGTTCTTGTCAATAAGGTACTGCTAGTTGAACAGCTCTTCC
  GCAAGGAGTTCCAACCATTTTTGAAGAAATGGTATCGTGTTATTGGATTAAGTGGTGATACCCAACTGAA
  AATATCATTTCCAGAAGTTGTCAAGTCCTGTGATATTATTATCAGTACAGCTCAAATCCTTGAAAACTCC
  CTCTTAAACTTGGAAAATGGAGAAGATGCTGGTGTTCAATTGTCAGACTTTTCCCTCATTATCATTGATG
  AATGTCATCACACCAACAAAGAAGCAGTGTATAATAACATCATGAGGCATTATTTGATGCAGAAGTTGAA
  AAACAATAGACTCAAGAAAGAAAACAAACCAGTGATTCCCCTTCCTCAGATACTGGGACTAACAGCTTCA
  CCTGGTGTTGGAGGGGCCACGAAGCAAGCCAAAGCTGAAGAACACATTTTAAAACTATGTGCCAATCTTG
  ATGCATTTACTATTAAAACTGTTAAAGAAAACCTTGATCAACTGAAAAACCAAATACAGGAGCCATGCAA
  GAAGTTTGCCATTGCAGATGCAACCAGAGAAGATCCATTTAAAGAGAAACTTCTAGAAATAATGACAAGG
  ATTCAAACTTATTGTCAAATGAGTCCAATGTCAGATTTTGGAACTCAACCCTATGAACAATGGGCCATTC
  AAATGGAAAAAAAAGCTGCAAAAGAAGGAAATCGCAAAGAACGTGTTTGTGCAGAACATTTGAGGAAGTA
  CAATGAGGCCCTACAAATTAATGACACAATTCGAATGATAGATGCGTATACTCATCTTGAAACTTTCTAT
  AATGAAGAGAAAGATAAGAAGTTTGCAGTCATAGAAGATGATAGTGATGAGGGTGGTGATGATGAGTATT
  GTGATGGTGATGAAGATGAGGATGATTTAAAGAAACCTTTGAAACTGGATGAAACAGATAGATTTCTCAT
  GACTTTATTTTTTGAAAACAATAAAATGTTGAAAAGGCTGGCTGAAAACCCAGAATATGAAAATGAAAAG
  CTGACCAAATTAAGAAATACCATAATGGAGCAATATACTAGGACTGAGGAATCAGCACGAGGAATAATCT
  TTACAAAAACACGACAGAGTGCATATGCGCTTTCCCAGTGGATTACTGAAAATGAAAAATTTGCTGAAGT
  AGGAGTCAAAGCCCACCATCTGATTGGAGCTGGACACAGCAGTGAGTTCAAACCCATGACACAGAATGAA
  CAAAAAGAAGTCATTAGTAAATTTCGCACTGGAAAAATAAATCTGCTTATCGCTACCACAGTGGCAGAAG
  AAGGTCTGGATATTAAAGAATGTAACATTGTTATCCGTTATGGTCTCGTCACCAATGAAATAGCCATGGT
  CCAGGCCCGTGGTCGAGCCAGAGCTGATGAGAGCACCTACGTCCTGGTTGCTCACAGTGGTTCAGGAGTT
  ATCGAACGTGAGACAGTTAATGATTTCCGAGAGAAGATGATGTATAAAGCTATACATTGTGTTCAAAATA
  TGAAACCAGAGGAGTATGCTCATAAGATTTTGGAATTACAGATGCAAAGTATAATGGAAAAGAAAATGAA
  AACCAAGAGAAATATTGCCAAGCATTACAAGAATAACCTATCACTAATAACTTTCCTTTGCAAAAACTGC
  AGTGTGCTAGCCTGTTCTGGGGAAGATATCCATGTAATTGAGAAAATGCATCACGTCAATATGACCCCAG
  AATTCAAGGAACTTTACATTGTAAGAGAAAACAAAACACTGCAAAAGAAGTGTGCCGACTATCAAATAAA
  TGGTGAAATCATCTGCAAATGTGGCCAGGCTTGGGGAACAATGATGGTGCACAAAGGCTTAGATTTGCCT
  TGTCTCAAAATAAGGAATTTTGTAGTGGTTTTCAAAAATAATTCAACAAAGAAACAATACAAAAAGTGGG
  TAGAATTACCTATCACATTTCCCAATCTTGACTATTCAGAATGCTGTTTATTTAGTGATGAGGAT
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC215661 representing NM_022168
  Red=Cloning site Green=Tags(s)
  
  MSNGYSTDENFRYLISCFRARVKMYIQVEPVLDYLTFLPAEVKEQIQRTVATSGNMQAVELLLSTLEKGV
  WHLGWTREFVEALRRTGSPLAARYMNPELTDLPSPSFENAHDEYLQLLNLLQPTLVDKLLVRDVLDKCME
  EELLTIEDRNRIAAAENNGNESGVRELLKRIVQKENWFSAFLNVLRQTGNNELVQELTGSDCSESNAEIE
  NLSQVDGPQVEEQLLSTTVQPNLEKEVWGMENNSSESSFADSSVVSESDTSLAEGSVSCLDESLGHNSNM
  GSDSGTMGSDSDEENVAARASPEPELQLRPYQMEVAQPALEGKNIIICLPTGSGKTRVAVYIAKDHLDKK
  KKASEPGKVIVLVNKVLLVEQLFRKEFQPFLKKWYRVIGLSGDTQLKISFPEVVKSCDIIISTAQILENS
  LLNLENGEDAGVQLSDFSLIIIDECHHTNKEAVYNNIMRHYLMQKLKNNRLKKENKPVIPLPQILGLTAS
  PGVGGATKQAKAEEHILKLCANLDAFTIKTVKENLDQLKNQIQEPCKKFAIADATREDPFKEKLLEIMTR
  IQTYCQMSPMSDFGTQPYEQWAIQMEKKAAKEGNRKERVCAEHLRKYNEALQINDTIRMIDAYTHLETFY
  NEEKDKKFAVIEDDSDEGGDDEYCDGDEDEDDLKKPLKLDETDRFLMTLFFENNKMLKRLAENPEYENEK
  LTKLRNTIMEQYTRTEESARGIIFTKTRQSAYALSQWITENEKFAEVGVKAHHLIGAGHSSEFKPMTQNE
  QKEVISKFRTGKINLLIATTVAEEGLDIKECNIVIRYGLVTNEIAMVQARGRARADESTYVLVAHSGSGV
  IERETVNDFREKMMYKAIHCVQNMKPEEYAHKILELQMQSIMEKKMKTKRNIAKHYKNNLSLITFLCKNC
  SVLACSGEDIHVIEKMHHVNMTPEFKELYIVRENKTLQKKCADYQINGEIICKCGQAWGTMMVHKGLDLP
  CLKIRNFVVVFKNNSTKKQYKKWVELPITFPNLDYSECCLFSDED
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_022168
• ORF Size: 3075 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_022168.4
• RefSeq Size: 3434 bp
• RefSeq ORF: 3078 bp
• Locus ID: 64135
• UniProt ID: Q9BYX4
• Cytogenetics: 2q24.2
• Domains: DEAD, helicase_C
• Protein Pathways: RIG-I-like receptor signaling pathway
• MW: 116.5 kDa
• Gene Summary: IFIH1 encodes MDA5 which is an intracellular sensor of viral RNA that triggers the innate immune response. Sensing RNA length and secondary structure, MDA5 binds dsRNA oligonucleotides with a modified DExD/H-box helicase core and a C-terminal domain, thus leading to a proinflammatory response that includes interferons. It has been shown that Coronaviruses (CoVs) as well as various other virus families, are capable of evading the MDA5-dependent interferon response, thus impeding the activation of the innate immune response to infection. MDA5 has also been shown to play an important role in enhancing natural killer cell function in malaria infection. In addition to its protective role in antiviral responses, MDA5 has been implicated in autoimmune and autoinflammatory diseases such as type 1 diabetes, systemic lupus erythematosus, and Aicardi-Goutieres syndrome[provided by RefSeq, Jul 2020]