LOCUS       sequence-463604-        1599 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of human RYBP NZF (20-51).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1599)
  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 1599)
  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-RYBP NZF (20-51)
FEATURES             Location/Qualifiers
     source          1..1599
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(40..57)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(177..200)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(219..872)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(239..261)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(873..890)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             903..925
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    940..964
                     /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(964..983)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(965..983)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1067..1086)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 actcagcttc tttcgggctt tgttagcagc cggatctcag tggtggtggt ggtggtgttt
       61 aaatggtcta gaaagcttta ggaggtgcct ttcctcacat cgcagatgct gcatttaaag
      121 gcttcagcac tgtttctgaa ggtgcagacg ctacaatccc aaaacccttc gtctgccggg
      181 ccctgaaaca gaacttccag accgctgctc agatccgatt ttggaggatg gtcgccacca
      241 ccaaacgtgg cttgccagcc ctgcaaaggc catgctatat acttgctgga tttcaagtac
      301 ttatcaattt gtgggatagc ttcaatacgt tttttaaaac aaactaattt tgggaacgca
      361 tccaggcaca ttgggtccat gtataaaaca acatcaagag cgtcatacaa catgaagtca
      421 ggatgggtta catgatcacc atttaaatat gttttatgac ataaacgatc ttcgaacatt
      481 ttcagcattt caggtagctt gctaagaaaa tcaactttga gagtttcaaa gtctttacta
      541 tatgcaattc tcgaaacacc gtatctaata tccaaaaccg ctccttcaag cattgaaatc
      601 tctgcacgct cttttggaca accacccaac atgttgtgct tgtcagctat ataacgtatg
      661 atggccatag actgtgttaa tttaacatca ccatcaatat aataaggaag attgggaaac
      721 tccaaaccca attcaaactt tttgtttcgc catttatcac cttcatcgcg ctcatacaaa
      781 tgctcttcat atttttcttc aagatattcc aaaagaagtc gagtgggttg cacaaggccc
      841 ttaattttcc aataacctag tataggggac atgtgatggt gatggtgatg tgccatggta
      901 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
      961 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga
     1021 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     1081 atcaccgatg gggaaaaatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt
     1141 gggtatggtg gcaggcccct tgccnggggg actgttgggn nccatctcct tgcatgcacc
     1201 atttctttnc gggggcgggg gctcaacggc ctcaacctac tactggggct gcttcctaat
     1261 gcaggaatnc cnnaannnaa aactccagaa tcccggaacc cattcaaagg gcccaaaacc
     1321 ttttcgcgna aggacttgaa agccccccga aaaagaatnn nttccggggg gggaaanngg
     1381 aaaaccntaa nngtttaaca gannccnaaa aaattgccgg nnttccttna aaannnnttt
     1441 tccccggnnn nnaaannnac ccnncnattt tttnnnnaaa accggggaaa annnnaaacg
     1501 ccgttggggg nnnnnnnntt tctcnnancc cgggnnanaa canatgggnn ngaaaanntn
     1561 tnngttaatg nnttcnnccc tcnanngttn antctnnna
//
