LOCUS       sequence-59530-e         907 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 907)
  AUTHORS   Nishi K, Nishi A, Nagasawa T, Ui-Tei K
  TITLE     Human TNRC6A is an Argonaute-navigator protein for microRNA-mediated
            gene silencing in the nucleus.
  JOURNAL   RNA. 2013 Jan;19(1):17-35. doi: 10.1261/rna.034769.112. Epub 2012 
            Nov 13.
  PUBMED    23150874
REFERENCE   2  (bases 1 to 907)
  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: 
            "10.1261/rna.034769.112"; journalName: "RNA"; date: "2013-01"; 
            volume: "19"; issue: "1"; pages: "17-35"
COMMENT     Sequence Label: pET-GST
FEATURES             Location/Qualifiers
     source          1..907
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(35..52)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(80..103)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(110..763)
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(130..152)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             complement(776..793)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(803..820)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             839..861
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    876..900
                     /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)."
ORIGIN
        1 cttcctttcg ggctttgtta gcagccggat ctcagtggtg gtggtggtgg tgctcgagtg
       61 cggccgcaag ctttcacagg ggcccctgga acagaacttc cagatccgat tttggaggat
      121 ggtcgccacc accaaacgtg gcttgccagc cctgcaaagg ccatgctata tacttgctgg
      181 atttcaagta cttatcaatt tgtgggatag cttcaatacg ttttttaaaa caaactaatt
      241 ttgggaacgc atccaggcac attgggtcca tgtataaaac aacatcaaga gcgtcataca
      301 acatgaagtc aggatgggtt acatgatcac catttaaata tgttttatga cataaacgat
      361 cttcgaacat tttcagcatt tcaggtagct tgctaagaaa atcaactttg agagtttcaa
      421 agtctttact atatgcaatt ctcgaaacac cgtatctaat atccaaaacc gctccttcaa
      481 gcattgaaat ctctgcacgc tcttttggac aaccacccaa catgttgtgc ttgtcagcta
      541 tataacgtat gatggccata gactgtgtta atttaacatc accatcaata taataaggaa
      601 gattgggaaa ctccaaaccc aattcaaact ttttgtttcg ccatttatca ccttcatcgc
      661 gctcatacaa atgctcttca tatttttctt caagatattc caaaagaagt cgagtgggtt
      721 gcacaaggcc cttaattttc caataaccta gtatagggga catgctagcc atatggctgc
      781 cgcgcggcac caggccgctg ctgtgatgat gatgatgatg gctgctgccc atggtatatc
      841 tccttcttaa agttaaacaa aattatttct agaggggaat tgttatccgc tcacaattcc
      901 cctatag
//
