LOCUS       sequence-110890-        1403 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses K29-/K33-specific TRABID NZF1 (aa 1-33) in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1403)
  AUTHORS   Michel MA, Elliott PR, Swatek KN, Simicek M, Pruneda JN, Wagstaff 
            JL, Freund SM, Komander D
  TITLE     Assembly and specific recognition of k29- and k33-linked 
            polyubiquitin.
  JOURNAL   Mol Cell. 2015 Apr 2;58(1):95-109. doi: 
            10.1016/j.molcel.2015.01.042. Epub 2015 Mar 5.
  PUBMED    25752577
REFERENCE   2  (bases 1 to 1403)
  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"; doi: 
            "10.1016/j.molcel.2015.01.042"; journalName: "Mol Cell"; date: 
            "2015-04-2- 2"; volume: "58"; issue: "1"; pages: "95-109"
COMMENT     Sequence Label: pOPINK-TRABID (NZF1, aa 1-33)
FEATURES             Location/Qualifiers
     source          1..1403
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(29..46)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(169..192)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(211..864)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(231..253)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(865..882)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             895..917
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    932..956
                     /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(956..975)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(957..975)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1059..1078)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tcttcgggct tntagcagcc ggatctcagt ggtggtggtg gtggtgttta aatggtctag
       61 aaagctttaa ctaggtcttt gggcacgaca catagtacac ttgattgcag atggccagtt
      121 ttcatacgta caatattcac aagcccactt aattccacgt tctgacatcg ggccctgaaa
      181 cagaacttcc agaccgctgc tcagatccga ttttggagga tggtcgccac caccaaacgt
      241 ggcttgccag ccctgcaaag gccatgctat atacttgctg gatttcaagt acttatcaat
      301 ttgtgggata gcttcaatac gttttttaaa acaaactaat tttgggaacg catccaggca
      361 cattgggtcc atgtataaaa caacatcaag agcgtcatac aacatgaagt caggatgggt
      421 tacatgatca ccatttaaat atgttttatg acataaacga tcttcgaaca ttttcagcat
      481 ttcaggtagc ttgctaagaa aatcaacttt gagagtttca aagtctttac tatatgcaat
      541 tctcgaaaca ccgtatctaa tatccaaaac cgctccttca agcattgaaa tctctgcacg
      601 ctcttttgga caaccaccca acatgttgtg cttgtcagct atataacgta tgatggccat
      661 agactgtgtt aatttaacat caccatcaat ataataagga agattgggaa actccaaacc
      721 caattcaaac tttttgtttc gccatttatc accttcatcg cgctcataca aatgctcttc
      781 atatttttct tcaagatatt ccaaaagaag tcgagtgggt tgcacaaggc ccttaatttt
      841 ccaataacct agtatagggg acatgtgatg gtgatggtga tgtgccatgg tatatctcct
      901 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
      961 tagtgagtcg tattaatttc gcgggatcga gatctcgatc ctctacgccg gacgcatcgt
     1021 ggccggcatc accggcgcca caggtgcggt tgctggcgcc tatatcgccg acatcaccga
     1081 tggggaagat cgggctcgcc acttcgggct catgagcgct tgtttcggcg tgggtatggt
     1141 ggcaggcccc gtggccgggg gactgttggn nnccatctcc ttgcatgnnc cattccttgc
     1201 ggnggnggng nncaacggnc tcaacctaat actggnctgg ttcctaatgc aggnntccna
     1261 taagggaanc ggccagatcc cggaaccntn caanggncca aaaccttncc gggatggnnt
     1321 gnnanncccg gaaangatca attcaggnng ggannnggaa acnannacgt tnacaanngc
     1381 aaanttagcg ggtgtctann aaa
//
