LOCUS       sequence-463605-        1123 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of human NPL4 NZF (574-608).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1123)
  AUTHORS   Michel MA, Scutts S, Komander D
  TITLE     Secondary interactions in ubiquitin-binding domains achieve linkage 
            or substrate specificity.
  JOURNAL   Cell Rep. 2024 Aug 27;43(8):114545. doi: 
            10.1016/j.celrep.2024.114545. Epub 2024 Jul 23.
  PUBMED    39052481
REFERENCE   2  (bases 1 to 1123)
  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: "Cell Rep.";
            date: "2024-08-27"; volume: "43(8):114545. doi"; pages: " 
            10.1016/j.celrep.2024.114545. Epub 2024 Jul 23"
COMMENT     Sequence Label: pOPINK-NPL4 NZF (574-608)
FEATURES             Location/Qualifiers
     source          1..1123
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(27..44)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(173..196)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(215..868)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(235..257)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(869..886)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    936..960
                     /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)."
     gap             1111..1120
                     /estimated_length=10
ORIGIN
        1 cnngctttgt tagcagccgg atctcagtgg tggtggtggt ggtgtttaaa tggtctagaa
       61 agctttaggt ccggggaagg ctgcacatct cgcagtggcc tgtgccaggc tggttcatga
      121 aggtgcagtg ctgacaagcc cacatggcag aggtagcagc gcgtgcagag cccgggccct
      181 gaaacagaac ttccagaccg ctgctcagat ccgattttgg aggatggtcg ccaccaccaa
      241 acgtggcttg ccagccctgc aaaggccatg ctatatactt gctggatttc aagtacttat
      301 caatttgtgg gatagcttca atacgttttt taaaacaaac taattttggg aacgcatcca
      361 ggcacattgg gtccatgtat aaaacaacat caagagcgtc atacaacatg aagtcaggat
      421 gggttacatg atcaccattt aaatatgttt tatgacataa acgatcttcg aacattttca
      481 gcatttcagg tagcttgcta agaaaatcaa ctttgagagt ttcaaagtct ttactatatg
      541 caattctcga aacaccgtat ctaatatcca aaaccgctcc ttcaagcatt gaaatctctg
      601 cacgctcttt tggacaacca cccaacatgt tgtgcttgtc agctatataa cgtatgatgg
      661 ccatagactg tgttaattta acatcaccat caatataata aggaagattg ggaaactcca
      721 aacccaattc aaactttttg tttcgccatt tatcaccttc atcgcgctca tacaaatgct
      781 cttcatattt ttcttcaaga tattccaaaa gaagtcgagt gggttgcaca aggcccttaa
      841 ttttccaata acctagtata ggggacatgt gatggtgatg gtgatgtgcc atggtatatc
      901 tccttcttaa agntaaacaa aattatttct agaggggaat tgttatccgc tcacaattcc
      961 cctatagtga gtcnnnnnna atttcnccgg gnncgannat ctcgatcctc tacgncngga
     1021 ngcatcgntg nnnggnntca cnnnngcnnn nnntgcggnt tgctngnnnc cnntntcnnn
     1081 nanntcncnn annnnggnng aatnggnntc nnnnnnnnnn ggg
//
