LOCUS       sequence-146972-         860 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacteria expression plasmid for Ubiquitin G76C with removable His6 
            tag by TEV protease.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 860)
  AUTHORS   Morgan MT, Haj-Yahya M, Ringel AE, Bandi P, Brik A, Wolberger C
  TITLE     Structural basis for histone H2B deubiquitination by the SAGA DUB 
            module.
  JOURNAL   Science. 2016 Feb 12;351(6274):725-8. doi: 10.1126/science.aac5681.
  PUBMED    26912860
REFERENCE   2  (bases 1 to 860)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2016-02-12- 12"; volume: "351"; 
            issue: "6274"; pages: "725-8"
COMMENT     Sequence Label: His6TEV-UbG76C
FEATURES             Location/Qualifiers
     source          1..860
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(19..243)
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRG"
     CDS             complement(253..273)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(295..312)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             331..353
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    368..392
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(392..411)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(393..411)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(495..514)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        720..797
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(817..836)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gcagccggat cctcaacaac cacgaagtct caacacaaga tgaagagtag actccttttg
       61 aatattgtag tcagacaaag tacgtccatc ttccagctgc ttgccagcaa agatcagtct
      121 ctgctgatca ggaggaattc cttccttatc ctggatcttg gcctttacat tttctatcgt
      181 atccgagggt tcaacctcga gggtgatggt cttccccgta agggttttca cgaaaatctg
      241 catcatatgg ctgccctgaa aatacaggtt ttcggtggtc ggaatatcat aatcgtgatg
      301 atgatgatga tggctgctgc ccatggtata tctccttctt aaagttaaac aaaattattt
      361 ctagagggga attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg
      421 gatcgagatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
      481 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
      541 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
      601 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
      661 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagatcccgg
      721 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
      781 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
      841 tctcttatca gaccgtttcc
//
