LOCUS       sequence-257443-        1099 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of His6-Ubc9-C93A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1099)
  AUTHORS   Hofmann R, Akimoto G, Wucherpfennig TG, Zeymer C, Bode JW
  TITLE     Lysine acylation using conjugating enzymes for site-specific 
            modification and ubiquitination of recombinant proteins.
  JOURNAL   Nat Chem. 2020 Nov;12(11):1008-1015. doi: 10.1038/s41557-020-0528-y.
            Epub 2020 Sep 14.
  PUBMED    32929246
REFERENCE   2  (bases 1 to 1099)
  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"; doi: 
            "10.1038/s41557-020-0528-y"; journalName: "Nat Chem"; date: 
            "2020-11"; volume: "12"; issue: "11"; pages: "1008-1015"
COMMENT     Sequence Label: pET28-His6-Ubc9-C93A
FEATURES             Location/Qualifiers
     source          1..1099
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             3..25
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             44..61
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             71..88
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             578..595
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(648..666)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      662..709
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(833..852)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1043..1064
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 attttgttta actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca
       61 cagcagcggc ctggtgccgc gcggcagcca tatgatgtcc ggaattgccc tctctcgctt
      121 agctcaggaa cgcaaagcgt ggcgcaaaga tcacccgttt gggtttgttg ccgtgcctac
      181 caaaaatcct gacggtacga tgaatcttat gaactgggaa tgcgcgatcc ctggcaagaa
      241 aggcacaccg tgggaaggcg gcttgttcaa gctgcgtatg ttgttcaaag atgattaccc
      301 gtcaagtccg cctaaatgta aattcgagcc gccgttattt catccaaacg tgtatcccag
      361 cggtacggtt gccctgagca tccttgaaga agacaaagat tggcgtcccg caattaccat
      421 caagcagatt ctgctcggca ttcaagagct gttaaatgaa ccaaacattc aggacccagc
      481 gcaggcagaa gcctatacta tctactgtca gaaccgtgta gagtatgaga agcgcgtccg
      541 tgcacaagcg aaaaagtttg ccccgtcgta actcgagcac caccaccacc accactgaga
      601 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
      661 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
      721 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      781 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      841 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      901 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      961 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
     1021 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
     1081 atctcggyct attcttttg
//
