LOCUS       sequence-463603-        1153 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of human Sharpin NZF (347-376).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1153)
  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 1153)
  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-Sharpin NZF (347-376)
FEATURES             Location/Qualifiers
     source          1..1153
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(50..67)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(178..201)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(220..873)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(240..262)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(874..891)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             904..926
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    941..965
                     /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(965..984)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(966..984)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gnnnnnnnnn actcagcttc tttcgggctt tgttagcagc cggatctcag tggtggtggt
       61 ggtggtgttt aaatggtcta gaaagcttta gggcctctgg gtgctacaca tctcacagcc
      121 agggcggtct ggggcattga tgaaggtgca ggaaggacag gaccagctgg gctggagtgg
      181 gccctgaaac agaacttcca gaccgctgct cagatccgat tttggaggat ggtcgccacc
      241 accaaacgtg gcttgccagc cctgcaaagg ccatgctata tacttgctgg atttcaagta
      301 cttatcaatt tgtgggatag cttcaatacg ttttttaaaa caaactaatt ttgggaacgc
      361 atccaggcac attgggtcca tgtataaaac aacatcaaga gcgtcataca acatgaagtc
      421 aggatgggtt acatgatcac catttaaata tgttttatga cataaacgat cttcgaacat
      481 tttcagcatt tcaggtagct tgctaagaaa atcaactttg agagtttcaa agtctttact
      541 atatgcaatt ctcgaaacac cgtatctaat atccaaaacc gctccttcaa gcattgaaat
      601 ctctgcacgc tcttttggac aaccacccaa catgttgtgc ttgtcagcta tataacgtat
      661 gatggccata gactgtgtta atttaacatc accatcaata taataaggaa gattgggaaa
      721 ctccaaaccc aattcaaact ttttgtttcg ccatttatca ccttcatcgc gctcatacaa
      781 atgctcttca tatttttctt caagatattc caaaagaagt cgagtgggtt gcacaaggcc
      841 cttaattttc caataaccta gtatagggga catgtgatgg tgatggtgat gtgccatggt
      901 atatctcctt cttaaagtta aacaaaatta tttctagagg ggaattgtta tccgctcaca
      961 attcccctat agtgagtcgt attaatttcg cgggatcgan atctcgatcc tctacgccgg
     1021 acgcatcgtg gnccggcatc accnggcgcc acagggggcg gttgctgggc gcctatatcg
     1081 cccaacatca ccgaaggggt aaaaatcggg nntcgccact ttcgggnctc atgaacctct
     1141 tgtttccggc tag
//
