LOCUS       sequence-463592-        1701 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of human CAPN15 NZF1 (1-42).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1701)
  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 1701)
  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 NZF1 (1-42)
FEATURES             Location/Qualifiers
     source          1..1701
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(32..49)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(198..221)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(240..893)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(260..282)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(894..911)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             924..946
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    961..985
                     /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(985..1004)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(986..1004)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1088..1107)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     gap             1573..1586
                     /estimated_length=14
     gap             1595..1604
                     /estimated_length=10
     gap             1607..1617
                     /estimated_length=11
ORIGIN
        1 gatnctcttn ggcttgtagc agccggatct cgtggtggtg gtggtggtgt ttaatggtct
       61 agaaagcttt agctgagccg caggatgtgg ttgaggtcgg gcttgtgccg gggagcctcg
      121 cagatggagc actggcgctg gccggccggg ttcaggaagg tgcagcgcac acaggaccac
      181 tctccgaccg tggccatcgg gccctgaaac agaacttcca gaccgctgct cagatccgat
      241 tttggaggat ggtcgccacc accaaacgtg gcttgccagc cctgcaaagg ccatgctata
      301 tacttgctgg atttcaagta cttatcaatt tgtgggatag cttcaatacg ttttttaaaa
      361 caaactaatt ttgggaacgc atccaggcac attgggtcca tgtataaaac aacatcaaga
      421 gcgtcataca acatgaagtc aggatgggtt acatgatcac catttaaata tgttttatga
      481 cataaacgat cttcgaacat tttcagcatt tcaggtagct tgctaagaaa atcaactttg
      541 agagtttcaa agtctttact atatgcaatt ctcgaaacac cgtatctaat atccaaaacc
      601 gctccttcaa gcattgaaat ctctgcacgc tcttttggac aaccacccaa catgttgtgc
      661 ttgtcagcta tataacgtat gatggccata gactgtgtta atttaacatc accatcaata
      721 taataaggaa gattgggaaa ctccaaaccc aattcaaact ttttgtttcg ccatttatca
      781 ccttcatcgc gctcatacaa atgctcttca tatttttctt caagatattc caaaagaagt
      841 cgagtgggtt gcacaaggcc cttaattttc caataaccta gtatagggga catgtgatgg
      901 tgatggtgat gtgccatggt atatctcctt cttaaagtta aacaaaatta tttctagagg
      961 ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag
     1021 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
     1081 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
     1141 atgagcgctt gtttncgcgt gggtatggtg gcaggccccg tggccggggn actgttgggc
     1201 gccanctcct tggatnnncc attccttggn ggngnngggg ntcaacggcc tcaacctaat
     1261 actggggctn nttcctaatg cannnnnccn nnaaaggaaa acntncannn nccccgaann
     1321 nnnncaanng ggccaaacct ttccnngnnn ngnnntnaaa nnncccggnn annnnnnnat
     1381 nnnggngnng gannnggaan cncnnaancg ttnncannnn ncnaaannnn nnnngtnnnc
     1441 cnnnnnaann cgttnnccnn ggnngannaa nnnncnnnnc tttntncnna aaancggnna
     1501 aaannnnnan nnngnnnggn gnannnnnnn nncnnncnnn ccgnnngnnn annannggnn
     1561 nnnnannnnn ctnnnnnnnn nnnnnngnnn ccccnnnnnn nnnnccnnnn nnnnnnnann
     1621 nnnnngnnng nnnnaantcc cnnnnnaaan annnngnnnn nnnnggnnnn nnagnnnnaa
     1681 naannnnnnn nnanacgnnn g
//
