LOCUS       sequence-463600-        1662 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of human CAPN15 ZF4 (337-369).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1662)
  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 1662)
  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-CAPN15 ZF4 (337-369)
FEATURES             Location/Qualifiers
     source          1..1662
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(33..50)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(172..195)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(214..867)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(234..256)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(868..885)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             898..920
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    935..959
                     /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(959..978)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(960..978)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1062..1081)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     gap             1476..1496
                     /estimated_length=21
     gap             1553..1564
                     /estimated_length=12
     gap             1623..1641
                     /estimated_length=19
ORIGIN
        1 gnngctcttn gggctngtag cagccggatc tcgtggtggt ggtggtggtg tttaatggtc
       61 tagaaagctt tagtgcagtt tggagcagcc gcaggccgag cacctggggg ccactgtggg
      121 gtttctgagc gtgcacttgg cacatgacca ggtggtgaag tcggggctgg acgggccctg
      181 aaacagaact tccagaccgc tgctcagatc cgattttgga ggatggtcgc caccaccaaa
      241 cgtggcttgc cagccctgca aaggccatgc tatatacttg ctggatttca agtacttatc
      301 aatttgtggg atagcttcaa tacgtttttt aaaacaaact aattttggga acgcatccag
      361 gcacattggg tccatgtata aaacaacatc aagagcgtca tacaacatga agtcaggatg
      421 ggttacatga tcaccattta aatatgtttt atgacataaa cgatcttcga acattttcag
      481 catttcaggt agcttgctaa gaaaatcaac tttgagagtt tcaaagtctt tactatatgc
      541 aattctcgaa acaccgtatc taatatccaa aaccgctcct tcaagcattg aaatctctgc
      601 acgctctttt ggacaaccac ccaacatgtt gtgcttgtca gctatataac gtatgatggc
      661 catagactgt gttaatttaa catcaccatc aatataataa ggaagattgg gaaactccaa
      721 acccaattca aactttttgt ttcgccattt atcaccttca tcgcgctcat acaaatgctc
      781 ttcatatttt tcttcaagat attccaaaag aagtcgagtg ggttgcacaa ggcccttaat
      841 tttccaataa cctagtatag gggacatgtg atggtgatgg tgatgtgcca tggtatatct
      901 ccttcttaaa gttaaacaaa attatttcta gaggggaatt gttatccgct cacaattccc
      961 ctatagtgag tcgtattaat ttcgcgggat cgagatctcg atcctctacg ccggacgcat
     1021 cgtggccggc atcaccggcg ccacaggtgc ggttgctggc gcctatatcg ccgacatcac
     1081 cgatggggga agatcgggct cgccacttcg ggctcatgaa cgcttggttc ggcgtgggta
     1141 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgnnt gnnccattcc
     1201 ttnnngggnc ggtgctcaan ggcctcaacc tannactggg nctgnttcct aatgnaggnn
     1261 nccnnnaaag ggaaannnnn caannncccc ggaaccnanc caantggncn naaanccttt
     1321 nnncgnnnnn nggcntnnan acnnncccgg naaaaaannn nccattnnng ggnngnnnnn
     1381 nnngggaann nccnnaancg ttnnnnnaag nnccccaaaa nnnnnngnnn gnnnctcnnn
     1441 nnnnaannnn gttnnccnnn gnnnnngann caangnnnnn nnnnnnnnnn nnnnnnanaa
     1501 nnnnnnnnna naaannnnna nnnnnnnaan ggnnnnannn naaannnnnn ncnnnnnnnn
     1561 nnnngnnaaa aannnggnnn nnnnanaanc tnnnnnnnnt aagnnnnnnn tcccccnnnn
     1621 ntnnnnnnnn nnnnnnnnnn ncnnnnnaan nngnnnnnnn na
//
