LOCUS       sequence-141571-         845 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of hRPN13 in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 845)
  AUTHORS   VanderLinden RT, Hemmis CW, Yao T, Robinson H, Hill CP
  TITLE     Structure and energetics of pairwise interactions between proteasome
            subunits RPN2, RPN13, and ubiquitin clarify a substrate recruitment 
            mechanism.
  JOURNAL   J Biol Chem. 2017 Apr 25. pii: jbc.M117.785287. doi: 
            10.1074/jbc.M117.785287.
  PUBMED    28442575
REFERENCE   2  (bases 1 to 845)
  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. 2017 Apr 25. pii: jbc.M117.785287. doi: 
            10.1074/jbc.M117.785287."
COMMENT     Sequence Label: pET151-HIS-TEV-GST-PP-RPN13
FEATURES             Location/Qualifiers
     source          1..845
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             2..24
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             35..52
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             53..94
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             95..115
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             116..766
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="SPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKKF
                     ELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRYG
                     VSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVLY
                     MDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     724..746
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             773..796
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 ttttgtttaa ctttaagaag gagatataca tatgcatcat caccatcacc atggtaagcc
       61 tatccctaac cctctcctcg gtctcgattc tacggaaaac ctgtattttc agggatcccc
      121 tatactaggt tattggaaaa ttaagggcct tgtgcaaccc actcgacttc ttttggaata
      181 tcttgaagaa aaatatgaag agcatttgta tgagcgcgat gaaggtgata aatggcgaaa
      241 caaaaagttt gaattgggtt tggagtttcc caatcttcct tattatattg atggtgatgt
      301 taaattaaca cagtctatgg ccatcatacg ttatatagct gacaagcaca acatgttggg
      361 tggttgtcca aaagagcgtg cagagatttc aatgcttgaa ggagcggttt tggatattag
      421 atacggtgtt tcgagaattg catatagtaa agactttgaa actctcaaag ttgattttct
      481 tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt ttatgtcata aaacatattt
      541 aaatggtgat catgtaaccc atcctgactt catgttgtat gacgctcttg atgttgtttt
      601 atacatggac ccaatgtgcc tggatgcgtt cccaaaatta gtttgtttta aaaaacgtat
      661 tgaagctatc ccacaaattg ataagtactt gaaatccagc aagtatatag catggccttt
      721 gcagggctgg caagccacgt ttggtggtgg cgaccatcct ccaaaatcgg atctggaagt
      781 tctgttccag gggccaatga cgacctcagg cgcgctcttt ccaagcctgg tgccaggctc
      841 tcggg
//
