LOCUS       sequence-166133-        1064 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  mammalian expression of ubiquitin-K48R-EGFP fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1064)
  AUTHORS   Tsirigotis M, Zhang M, Chiu RK, Wouters BG, Gray DA
  TITLE     Sensitivity of mammalian cells expressing mutant ubiquitin to 
            protein-damaging agents.
  JOURNAL   J Biol Chem. 2001 Dec 7;276(49):46073-8.
  PUBMED    11598140
REFERENCE   2  (bases 1 to 1064)
  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 Biol 
            Chem."; date: "2001-12-7"; volume: "276(49)"; pages: "46073-8"
COMMENT     Sequence Label: pDG279
FEATURES             Location/Qualifiers
     source          1..1064
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             135..152
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             153..380
                     /codon_start=1
                     /product="human ubiquitin variant in which Lys-48 has been 
                     mutated to arginine"
                     /label=ubiquitin-K48R
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGRQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             join(381..383,384..386,387..899)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(426..447)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(687..706)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1034..1055
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 agttttttag gcaccttttg aaatgtaatc atttgggtca atatgtaatt ttcagtgtta
       61 gactagtaaa ttgtccgcta aattctggcc gtttttggct tttttgttag acgaagctta
      121 aaatgagagg cagccaccac catcaccatc acatgcagat cttcgtgaag accctgaccg
      181 gcaagaccat cactctggag gtggagccca gtgacaccat cgaaaatgtg aaggccaaga
      241 tccaagataa agaaggcatc ccccccgacc agcagaggct catctttgca ggccggcagc
      301 tggaagatgg ccgtactctt tctgactaca acatccagaa agagtcgacc ctgcacctgg
      361 tcctgcgtct gagaggtggt atggtgagca agggcgagga gctgttcacc ggggtggtgc
      421 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
      481 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
      541 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
      601 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
      661 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
      721 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
      781 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
      841 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
      901 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      961 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     1021 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgcc
//
