LOCUS       sequence-208269-        1182 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of Met1 diUb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1182)
  AUTHORS   Swatek KN, Aumayr M, Pruneda JN, Visser LJ, Berryman S, Kueck AF, 
            Geurink PP, Ovaa H, van Kuppeveld FJM, Tuthill TJ, Skern T, Komander
            D
  TITLE     Irreversible inactivation of ISG15 by a viral leader protease 
            enables alternative infection detection strategies.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Mar 6;115(10):2371-2376. doi: 
            10.1073/pnas.1710617115. Epub 2018 Feb 20.
  PUBMED    29463763
REFERENCE   2  (bases 1 to 1182)
  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: 
            "10.1073/pnas.1710617115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-03-6- 6"; volume: "115"; issue: "10"; pages: "2371-2376"
COMMENT     Sequence Label: Met1 diUb (1-152, R42W)
FEATURES             Location/Qualifiers
     source          1..1182
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             8..30
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             49..66
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             76..99
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             100..327
                     /codon_start=1
                     /gene="Homo sapiens UBC"
                     /product="human ubiquitin C"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQWLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             328..555
                     /codon_start=1
                     /product="human ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVEPSDTIENVKAKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     CDS             582..599
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(652..670)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      666..713
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      750..1175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(837..856)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1046..1067
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gaataatttt gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat
       61 catcacagca gcggtctgga agttctgttt cagggcccga tgcagatctt cgtgaagacc
      121 ctgactggta agaccatcac tctcgaagtg gagccgagtg acaccattga gaatgtcaag
      181 gcaaagatcc aagacaagga aggcatccct cctgaccagc agtggttgat ctttgctggg
      241 aagcagctgg aagatggacg caccctgtct gactacaaca tccagaaaga gtccaccctg
      301 cacctggtcc tccgtctcag aggtgggatg caaatatttg tcaaaacact aacaggaaaa
      361 acaataacac tagaggtcga accctctgat acaatagaaa acgtgaaagc caaaatacag
      421 gataaagagg gaataccccc cgatcaacaa aggctaatat tcgccggaaa acaactagag
      481 gacgggagga cactaagtga ttataatata caaaaggaaa gtacactaca tctagtgcta
      541 aggttaagag gtgggtgata aagctttcta gaccatttaa acaccaccac caccaccact
      601 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      661 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
      721 gaggaactat atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc
      781 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
      841 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
      901 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaactt
      961 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     1021 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     1081 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     1141 aaaatgagct gatttaacaa aatttaacgc gaattttaac aa
//
