LOCUS       sequence-463593-        1339 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of human CAPN15 NZF2 (34-72).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1339)
  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 1339)
  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 NZF2 (34-72)
FEATURES             Location/Qualifiers
     source          1..1339
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(39..56)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(202..225)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(244..897)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(264..286)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(898..915)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             928..950
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    965..989
                     /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(989..1008)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(990..1008)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1092..1111)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 aaggggnann cncttngggc tnntnacgcc ggatctcagt ggtggtggtg gtggtgttta
       61 atggtctaga aagctttacg caggctccgg ggtgaagccg cacacctcgc aggcctcctt
      121 gcccaggaag ttgcggaagg tgcagcgggc gcagggccat ttctgctcct ccacgctgag
      181 ccgcaggatg tggttgaggt ccgggccctg aaacagaact tccagaccgc tgctcagatc
      241 cgattttgga ggatggtcgc caccaccaaa cgtggcttgc cagccctgca aaggccatgc
      301 tatatacttg ctggatttca agtacttatc aatttgtggg atagcttcaa tacgtttttt
      361 aaaacaaact aattttggga acgcatccag gcacattggg tccatgtata aaacaacatc
      421 aagagcgtca tacaacatga agtcaggatg ggttacatga tcaccattta aatatgtttt
      481 atgacataaa cgatcttcga acattttcag catttcaggt agcttgctaa gaaaatcaac
      541 tttgagagtt tcaaagtctt tactatatgc aattctcgaa acaccgtatc taatatccaa
      601 aaccgctcct tcaagcattg aaatctctgc acgctctttt ggacaaccac ccaacatgtt
      661 gtgcttgtca gctatataac gtatgatggc catagactgt gttaatttaa catcaccatc
      721 aatataataa ggaagattgg gaaactccaa acccaattca aactttttgt ttcgccattt
      781 atcaccttca tcgcgctcat acaaatgctc ttcatatttt tcttcaagat attccaaaag
      841 aagtcgagtg ggttgcacaa ggcccttaat tttccaataa cctagtatag gggacatgtg
      901 atggtgatgg tgatgtgcca tggtatatct ccttcttaaa gttaaacaaa attatttcta
      961 gaggggaatt gttatccgct cacaattccc ctatagtgag tcgtattaat ttcgcgggat
     1021 cgagatctcg atcctctacg ccggacgcat cgtggccggc atcaccggcg ccacaggtgc
     1081 ggttgctggc gcctatatcg ccgacatcac cgatggggaa gatcgggctc gccacttcgg
     1141 gctcatgaac gcttgtttcg gcgtgggtat ggtggcaggc cccgtggccg ggggactgtt
     1201 gggcgccatc tccttggatg gcccattcct tgggggggng nnggtnaacg gcctcaacct
     1261 aatactggnn nngcttccta atgcaggant ccnataaggg aaancgtccn nnncccggaa
     1321 ccctccaang ggnnaaanc
//
