LOCUS       sequence-108955-         874 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression of HA tagged ubiquitin carrying zero lysines 
            (K0).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 874)
  AUTHORS   Lim KL, Chew KC, Tan JM, Wang C, Chung KK, Zhang Y, Tanaka Y, Smith 
            W, Engelender S, Ross CA, Dawson VL, Dawson TM
  TITLE     Parkin mediates nonclassical, proteasomal-independent ubiquitination
            of synphilin-1: implications for Lewy body formation.
  JOURNAL   J Neurosci. 2005 Feb 23. 25(8):2002-9.
  PUBMED    15728840
REFERENCE   2  (bases 1 to 874)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Neurosci.
            2005 Feb 23. 25(8):2002-9."
COMMENT     Sequence Label: pRK5-HA-Ubiquitin-KO
FEATURES             Location/Qualifiers
     source          1..874
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        228..246
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     228..245
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     259..278
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      363..372
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             375..401
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             432..659
                     /codon_start=1
                     /product="human ubiquitin variant in which all of the 
                     lysines have been mutated to arginine"
                     /label=ubiquitin-K0
                     /note="also known as ubiquitin-KO"
                     /translation="MQIFVRTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQRESTLHLVLRLRGG"
     regulatory      696..705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    709..843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(746..765)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     800..819
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ttagtgaacc gtcagatcgc ctggagacgc catccacgct gttttgacct ccatagaaga
       61 caccgggacc gatccagcct ccgcggccgg gaacggtgca ttggaacgcg gattccccgt
      121 gccaagagtg acgtaagtac cgcctataga gtctataggc ccaccccctt ggcttcgtta
      181 gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt taggtgacac
      241 tatagaataa catccacttt gcctttctct ccacaggtgt ccactcccag gtccaactgc
      301 acctcggttc tatcgattga attcccgaac cgacagtcgg tctcttcacc aaggccattc
      361 gcgccaccat gggctacccc tatgatgtgc ctgactacgc agatctcaat ggtggtggtg
      421 gtgggtcgac catgcagatc ttcgtcagaa cgttaaccgg tagaaccata actctagaag
      481 ttgaaccatc cgataccatc gaaaacgtta gagctagaat tcaagacaga gaaggcattc
      541 cacctgatca acaaagattg atctttgccg gtagacagct cgaggacggt agaacgctgt
      601 ctgattacaa cattcagaga gagtcgacct tacatcttgt cttaagacta agaggtggtt
      661 gagcggccgc taagtaagta aggatcccca gcttggccgc catggcccaa cttgtttatt
      721 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
      781 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg
      841 atcgggaatt aattcggcgc agcaccatgg cctg
//
