LOCUS       sequence-463595-        1381 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of human CAPN15 ZF3 (141-173).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1381)
  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 1381)
  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 ZF3 (141-173)
FEATURES             Location/Qualifiers
     source          1..1381
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(36..53)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(175..198)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(217..870)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(237..259)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(871..888)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             901..923
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    938..962
                     /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(962..981)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(963..981)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1065..1084)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggnnnnctct tcgggcttgt agcagccgga tctcagtggt ggtggtggtg gtgttaaatg
       61 gtctagaaag ctttacgaga gcctgcgtgg gcccccgcag acggagcagg agctggccac
      121 gggcgtgttg tgcagcgtgc aacgcggaca cgcccagccc cctctgggct ccgccgggcc
      181 ctgaaacaga acttccagac cgctgctcag atccgatttt ggaggatggt cgccaccacc
      241 aaacgtggct tgccagccct gcaaaggcca tgctatatac ttgctggatt tcaagtactt
      301 atcaatttgt gggatagctt caatacgttt tttaaaacaa actaattttg ggaacgcatc
      361 caggcacatt gggtccatgt ataaaacaac atcaagagcg tcatacaaca tgaagtcagg
      421 atgggttaca tgatcaccat ttaaatatgt tttatgacat aaacgatctt cgaacatttt
      481 cagcatttca ggtagcttgc taagaaaatc aactttgaga gtttcaaagt ctttactata
      541 tgcaattctc gaaacaccgt atctaatatc caaaaccgct ccttcaagca ttgaaatctc
      601 tgcacgctct tttggacaac cacccaacat gttgtgcttg tcagctatat aacgtatgat
      661 ggccatagac tgtgttaatt taacatcacc atcaatataa taaggaagat tgggaaactc
      721 caaacccaat tcaaactttt tgtttcgcca tttatcacct tcatcgcgct catacaaatg
      781 ctcttcatat ttttcttcaa gatattccaa aagaagtcga gtgggttgca caaggccctt
      841 aattttccaa taacctagta taggggacat gtgatggtga tggtgatgtg ccatggtata
      901 tctccttctt aaagttaaac aaaattattt ctagagggga attgttatcc gctcacaatt
      961 cccctatagt gagtcgtatt aatttcgcgg gatcgagatc tcgatcctct acgccggacg
     1021 catcgtggcc ggcatcaccg gcgccacagg tgcggttgct ggcgcctata tcgccgacat
     1081 caccgatggg gaagatcggg ctcgccactt cgggctcatg aacgcttgtt tcggcgtggg
     1141 tatggtggca ggccccgtgc cnggngnact gttgggcgcc atctccttgc atgcaccatt
     1201 ccttgnggng ncggnggtca acggcctcaa cctactactg ggnttgcttc ctaatnnaan
     1261 natcccataa agggaaancn nncaagatcc cggacaccat ccaannggnn naaaaccttt
     1321 ccnggnnnng nnnngaaacc ccccggaaaa angtcaatnn ngggggggga anngganacc
     1381 c
//
