LOCUS       sequence-164012-        1003 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 1003)
  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 1003)
  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..1003
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(31..48)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(123..350)
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             complement(363..383)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(405..422)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             442..464
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    479..503
                     /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(503..522)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(504..522)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(606..625)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        831..908
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(928..947)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ctttcgggct ttgttagcag ccggatctca gtggtggtgg tggtggtgct cgagactgca
       61 ggctctagat tcgaaagcgg ccgcgactag tgagctcgtc gacgtaggcc tttgaattct
      121 cacccacctc tgagacggag taccaggtgc aaggtggact ctttctggat gttgtagtca
      181 gacagggtac gaccatcttc cagctgtttt ccggcaaaga tcaacctctg ctgatcagga
      241 ggaatgcctt ccttatcttg gatctttgcc ttgacattct cgatggtgtc actgggctcc
      301 acctcgaggg tgatggtctt accagtcaga gtcttcacga agatctgcat ggatcccatg
      361 gcgccctgaa aatacaggtt ttcggtcgtt gggatatcgt aatcgtgatg gtgatggtga
      421 tggtagtacg acatatgtat atctccttct taaagttaaa caaaattatt tctagagggg
      481 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
      541 ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
      601 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgggctcat
      661 gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg gccgggggac tgttgggcgc
      721 catctccttg catgcaccat tccttgcggc ggcggtgctc aacggcctca acctactact
      781 gggctgcttc ctaatgcagg agtcgcataa gggagagcgt cgagatcccg gacaccatcg
      841 aatggcgcaa aacctttcgc ggtatggcat gatagcgccc ggaagagagt caattcaggg
      901 tggtgaatgt gaaaccagta acgttatacg atgtcgcaga gtatgccggt gtctcttatc
      961 agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc tgc
//
