LOCUS       sequence-463601-        1545 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of human CAPN15 ZF5 (411-444).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1545)
  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 1545)
  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 ZF5 (411-444)
FEATURES             Location/Qualifiers
     source          1..1545
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(34..51)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(176..199)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(218..871)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(238..260)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(872..889)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             902..924
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    939..963
                     /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(963..982)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(964..982)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1066..1085)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggatgctctt cgggcttgta gcagccggat ctcgtggtgg tggtggtggt gtttaatggt
       61 ctagaaagct ttactgggcc accaggagct gaggcgtgtg gcaggcggcg cagtgcttgg
      121 cccgcagtgc gttgagcagg gtacaggcag ggcaggccca ctggcccggg cgctccgggc
      181 cctgaaacag aacttccaga ccgctgctca gatccgattt tggaggatgg tcgccaccac
      241 caaacgtggc ttgccagccc tgcaaaggcc atgctatata cttgctggat ttcaagtact
      301 tatcaatttg tgggatagct tcaatacgtt ttttaaaaca aactaatttt gggaacgcat
      361 ccaggcacat tgggtccatg tataaaacaa catcaagagc gtcatacaac atgaagtcag
      421 gatgggttac atgatcacca tttaaatatg ttttatgaca taaacgatct tcgaacattt
      481 tcagcatttc aggtagcttg ctaagaaaat caactttgag agtttcaaag tctttactat
      541 atgcaattct cgaaacaccg tatctaatat ccaaaaccgc tccttcaagc attgaaatct
      601 ctgcacgctc ttttggacaa ccacccaaca tgttgtgctt gtcagctata taacgtatga
      661 tggccataga ctgtgttaat ttaacatcac catcaatata ataaggaaga ttgggaaact
      721 ccaaacccaa ttcaaacttt ttgtttcgcc atttatcacc ttcatcgcgc tcatacaaat
      781 gctcttcata tttttcttca agatattcca aaagaagtcg agtgggttgc acaaggccct
      841 taattttcca ataacctagt ataggggaca tgtgatggtg atggtgatgt gccatggtat
      901 atctccttct taaagttaaa caaaattatt tctagagggg aattgttatc cgctcacaat
      961 tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac
     1021 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     1081 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gaacgcttgt tncggcgtgg
     1141 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgnnccat
     1201 tccttnnngn ggcggggntc aacggcctca acctactact gggcnnnttc ctaatgcagn
     1261 aatccnnnaa agggaaancg tccannnncc cggaaccnnn ngaannggnn naaaaccttt
     1321 nnnngnnnng gnnngaaaac cccccgnaaa aannnnannn nngggnnnng gnannggaaa
     1381 ccntnanncg tnnnccaann nnccnaaann nngnnggggn tcnnttnnaa nnnnttnccc
     1441 ccgnnngnaa annnnnnnnn cnnttttnnc gnaannnnng nnaaaannnn nnnacnnnnn
     1501 nngnnnnnan antntnntcn nnnancnnnn nggaannann nnngg
//
