LOCUS       sequence-45166-e         685 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 685)
  AUTHORS   Sims JJ, Scavone F, Cooper EM, Kane LA, Youle RJ, Boeke JD, Cohen RE
  TITLE     Polyubiquitin-sensor proteins reveal localization and linkage-type 
            dependence of cellular ubiquitin signaling.
  JOURNAL   Nat Methods. 2012 Feb 5. doi: 10.1038/nmeth.1888.
  PUBMED    22306808
REFERENCE   2  (bases 1 to 685)
  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"; journalName: "Nat 
            Methods. 2012 Feb 5. doi: 10.1038/nmeth.1888."
COMMENT     Sequence Label: pET28a-Vx3(A7)
FEATURES             Location/Qualifiers
     source          1..685
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(28..45)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(320..337)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(347..364)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             383..405
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    420..444
                     /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(444..463)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(445..463)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(547..566)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcgggctttg ttagcagccg gatctcagtg gtggtggtgg tggtgctcga gtgcggccgc
       61 aagcttgtcg acggagctcg aattcggatc ctcaagcaga gttacggcac tccttcaggc
      121 tcagctcgat ggccttgcgg atcagctcct cctcgtcctc ggccgcggct gcagccgccg
      181 cagattcttt cagagacagt tcgatagctt tacggatcag ttcttcttcg tcttcagcgg
      241 cagccgccgc tgcggcagct tcacgcagag attcctggat agcagctttt aaatcaggat
      301 cttcttcttc ttccatatgg ctgccgcgcg gcaccaggcc gctgctgtga tgatgatgat
      361 gatggctgct gggcatggta tatctccttc ttaaagttaa acaaaattat ttctagaggg
      421 gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga
      481 tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
      541 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
      601 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
      661 ccatctcctt gcatgcacca ttcct
//
