LOCUS       sequence-360486-        1104 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of His6-Ubc9-K14R.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1104)
  AUTHORS   Akimoto G, Fernandes AP, Bode JW
  TITLE     Site-Specific Protein Ubiquitylation Using an Engineered, Chimeric 
            E1 Activating Enzyme and E2 SUMO Conjugating Enzyme Ubc9.
  JOURNAL   ACS Cent Sci. 2022 Feb 23;8(2):275-281. doi: 
            10.1021/acscentsci.1c01490. Epub 2022 Feb 9.
  PUBMED    35237717
REFERENCE   2  (bases 1 to 1104)
  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: 
            "10.1021/acscentsci.1c01490"; journalName: "ACS Cent Sci"; date: 
            "2022-02-23- 23"; volume: "8"; issue: "2"; pages: "275-281"
COMMENT     Sequence Label: pET28-His6-Ubc9-K14R
FEATURES             Location/Qualifiers
     source          1..1104
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             4..26
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             45..62
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             72..89
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             579..596
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(649..667)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      663..710
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      747..1104
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(834..853)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1044..1065
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aattttgttt aactttaaga aggagatata ccatgggcag cagccatcat catcatcatc
       61 acagcagcgg cctggtgccg cgcggcagcc atatgatgtc cggaattgcc ctctctcgct
      121 tagctcagga acgccgtgcg tggcgcaaag atcacccgtt tgggtttgtt gccgtgccta
      181 ccaaaaatcc tgacggtacg atgaatctta tgaactggga atgcgcgatc cctggcaaga
      241 aaggcacacc gtgggaaggc ggcttgttca agctgcgtat gttgttcaaa gatgattacc
      301 cgtcaagtcc gcctaaatgt aaattcgagc cgccgttatt tcatccaaac gtgtatccca
      361 gcggtacggt ttgcctgagc atccttgaag aagacaaaga ttggcgtccc gcaattacca
      421 tcaagcagat tctgctcggc attcaagagc tgttaaatga accaaacatt caggacccag
      481 cgcaggcaga agcctatact atctactgtc agaaccgtgt agagtatgag aagcgcgtcc
      541 gtgcacaagc gaaaaagttt gccccgtcgt aactcgagca ccaccaccac caccactgag
      601 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
      661 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
      721 gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg
      781 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
      841 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
      901 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
      961 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1021 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1081 tatctcggtc tattcttttg attt
//
