LOCUS       sequence-45489-e         937 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 937)
  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 937)
  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-Rx3(A7)
FEATURES             Location/Qualifiers
     source          1..937
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             7..29
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             48..65
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             75..92
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             358..375
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(428..446)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      442..489
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      526..906
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(613..632)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     823..844
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aataattttg tttaacttta agaaggagat ataccatgcc cagcagccat catcatcatc
       61 atcacagcag cggcctggtg ccgcgcggca gccatatgac cgaggaggag caattcgcac
      121 tcgcactgaa gatgagcgag caagaggcgg cggctgccgc tgctgccgca gaagaggaag
      181 aagaacagtt tgctctggct ctcaaaatga gtgaacagga agctgctgcc gcagcggcgg
      241 ctgccgagga ggaagaagag ctcttgagga aagccattgc agaaagtctg aatagttgcc
      301 ggccttcttg aggatccgaa ttcgagctcc gtcgacaagc ttgcggccgc actcgagcac
      361 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
      421 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
      481 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
      541 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      601 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      661 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      721 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
      781 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      841 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
      901 tttcggccta ttggttaaaa aatgagctga tttacaa
//
