LOCUS       sequence-164014-         939 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pHis-par2-Ub Hs.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 939)
  AUTHORS   Levin-Kravets O, Tanner N, Shohat N, Attali I, Keren-Kaplan T, 
            Shusterman A, Artzi S, Varvak A, Reshef Y, Shi X, Zucker O, Baram T,
            Katina C, Pilzer I, Ben-Aroya S, Prag G
  TITLE     A bacterial genetic selection system for ubiquitylation cascade 
            discovery.
  JOURNAL   Nat Methods. 2016 Nov;13(11):945-952. doi: 10.1038/nmeth.4003. Epub 
            2016 Oct 3.
  PUBMED    27694912
REFERENCE   2  (bases 1 to 939)
  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/nmeth.4003"; journalName: "Nat Methods"; date: "2016-11"; 
            volume: "13"; issue: "11"; pages: "945-952"
COMMENT     Sequence Label: pGEN25-Ub Hs
FEATURES             Location/Qualifiers
     source          1..939
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     266..285
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     369..388
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        369..387
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    388..412
                     /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)."
     RBS             427..449
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             469..486
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             508..528
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             541..768
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             843..860
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(913..931)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 actctcttcc gggcgctatc atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc
       61 cgggatctcg acgctctccc ttatgcgact cctgcattag gaagcagccc agtagtaggt
      121 tgaggccgtt gagcaccgcc gccgcaagga atggtgcatg caaggagatg gcgcccaaca
      181 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
      241 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
      301 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
      361 cgcgaaatta atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga
      421 aataattttg tttaacttta agaaggagat atacatatgt cgtactacca tcaccatcac
      481 catcacgatt acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catgggatcc
      541 atgcagatct tcgtgaagac tctgactggt aagaccatca ccctcgaggt ggagcccagt
      601 gacaccatcg agaatgtcaa ggcaaagatc caagataagg aaggcattcc tcctgatcag
      661 cagaggttga tctttgccgg aaaacagctg gaagatggtc gtaccctgtc tgactacaac
      721 atccagaaag agtccacctt gcacctggta ctccgtctca gaggtgggtg agaattcaaa
      781 ggcctacgtc gacgagctca ctagtcgcgg ccgctttcga atctagagcc tgcagtctcg
      841 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
      901 tggctgctgc caccgctgag caataactag cataacccc
//
