Apg3 (ATG3) (NM_022488) Human Tagged ORF Clone

CAT#: RC203453

ATG3 (Myc-DDK-tagged)-Human ATG3 autophagy related 3 homolog (S. cerevisiae) (ATG3)

Product Images

Circular map for RC203453

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Left lane) or pCMV6-ENTRY ATG3 (RC203453, 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-ATG3 (TA503346). Positive lysates LY411559 (100ug) and LC411559 (20ug) can be purchased separately from OriGene.

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  HEK293T cells were transfected with the pCMV6-ENTRY control (Cat# PS100001, Left lane) or pCMV6-ENTRY ATG3 (Cat# RC203453, 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-ATG3(Cat# TA503161). Positive lysates LY411559 (100ug) and LC411559 (20ug) can be purchased separately from OriGene.

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  Western blot validation of overexpression lysate (Cat# LY411559) using anti-DDK antibody (Cat# TA50011-100). Left: Cell lysates from un-transfected HEK293T cells; Right: Cell lysates from HEK293T cells transfected with RC203453 using transfection reagent MegaTran 2.0 (Cat# TT210002).

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  Coomassie blue staining of purified ATG3 protein (Cat# TP303453). The protein was produced from HEK293T cells transfected with ATG3 cDNA clone (Cat# RC203453) using MegaTran 2.0 (Cat# TT210002).

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  Anti-ATG3 mouse monoclonal antibody (TA503346) immunofluorescent staining of COS7 cells transiently transfected by pCMV6-ENTRY ATG3 (RC203453).

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Specifications

Product Data

• Type: Human Tagged ORF Clone
• Tag: Myc-DDK
• Symbol: Apg3
• Synonyms: APG3; APG3-LIKE; APG3L; PC3-96
• Vector: pCMV6-Entry
• E. coli Selection: Kanamycin (25 ug/mL)
• Mammalian Cell Selection: Neomycin
• Sequence Data:
  ORF Nucleotide Sequence

  >RC203453 ORF sequence
  Red=Cloning site Blue=ORF Green=Tags(s)
  
  TTTTGTAATACGACTCACTATAGGGCGGCCGGGAATTCGTCGACTGGATCCGGTACCGAGGAGATCTGCC
  GCCGCGATCGCC
  
  ATGCAGAATGTGATTAATACTGTGAAGGGAAAGGCACTGGAAGTGGCTGAGTACCTGACCCCGGTCCTCA
  AGGAATCAAAGTTTAAGGAAACAGGTGTAATTACCCCAGAAGAGTTTGTGGCAGCTGGAGATCACCTAGT
  CCACCACTGTCCAACATGGCAATGGGCTACAGGGGAAGAATTGAAAGTGAAGGCATACCTACCAACAGGC
  AAACAATTTTTGGTAACCAAAAATGTGCCGTGCTATAAGCGGTGCAAACAGATGGAATATTCAGATGAAT
  TGGAAGCTATCATTGAAGAAGATGATGGTGATGGCGGATGGGTAGATACATATCACAACACAGGTATTAC
  AGGAATAACGGAAGCCGTTAAAGAGATCACACTGGAAAATAAGGACAATATAAGGCTTCAAGATTGCTCA
  GCACTATGTGAAGAGGAAGAAGATGAAGATGAAGGAGAAGCTGCAGATATGGAAGAATATGAAGAGAGTG
  GATTGTTGGAAACAGATGAGGCTACCCTAGATACAAGGAAAATAGTAGAAGCTTGTAAAGCCAAAACTGA
  TGCTGGCGGTGAAGATGCTATTTTGCAAACCAGAACTTATGACCTTTACATCACTTATGATAAATATTAC
  CAGACTCCACGATTATGGTTGTTTGGCTATGATGAGCAACGGCAGCCTTTAACAGTTGAGCACATGTATG
  AAGACATCAGTCAGGATCATGTGAAGAAAACAGTGACCATTGAAAATCACCCTCATCTGCCACCACCTCC
  CATGTGTTCAGTTCACCCATGCAGGCATGCTGAGGTGATGAAGAAAATCATTGAGACTGTTGCAGAAGGA
  GGGGGAGAACTTGGAGTTCATATGTATCTTCTTATTTTCTTGAAATTTGTACAAGCTGTCATTCCAACAA
  TAGAATATGACTACACAAGACACTTCACAATG
  
  ACGCGTACGCGGCCGCTCGAGCAGAAACTCATCTCAGAAGAGGATCTGGCAGCAAATGATATCCTGGATT
  ACAAGGATGACGACGATAAGGTTTAA
  Protein Sequence

  >RC203453 protein sequence
  Red=Cloning site Green=Tags(s)
  
  MQNVINTVKGKALEVAEYLTPVLKESKFKETGVITPEEFVAAGDHLVHHCPTWQWATGEELKVKAYLPTG
  KQFLVTKNVPCYKRCKQMEYSDELEAIIEEDDGDGGWVDTYHNTGITGITEAVKEITLENKDNIRLQDCS
  ALCEEEEDEDEGEAADMEEYEESGLLETDEATLDTRKIVEACKAKTDAGGEDAILQTRTYDLYITYDKYY
  QTPRLWLFGYDEQRQPLTVEHMYEDISQDHVKKTVTIENHPHLPPPPMCSVHPCRHAEVMKKIIETVAEG
  GGELGVHMYLLIFLKFVQAVIPTIEYDYTRHFTM
  
  TRTRPLEQKLISEEDLAANDILDYKDDDDKV
• Restriction Sites: SgfI-MluI Cloning Scheme for this gene Plasmid Map
• ACCN: NM_022488
• ORF Size: 942 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_022488.5
• RefSeq Size: 1572 bp
• RefSeq ORF: 945 bp
• Locus ID: 64422
• UniProt ID: Q9NT62
• Cytogenetics: 3q13.2
• Domains: Autophagy_N, Autophagy_C
• Protein Pathways: Regulation of autophagy
• MW: 35.9 kDa
• Gene Summary: This gene encodes a ubiquitin-like-conjugating enzyme and is a component of ubiquitination-like systems involved in autophagy, the process of degradation, turnover and recycling of cytoplasmic constituents in eukaryotic cells. This protein is known to play a role in regulation of autophagy during cell death. A pseudogene of this gene is located on chromosome 20. Alternative splicing results in multiple transcript variants encoding different isoforms. [provided by RefSeq, Jul 2013]