LOCUS       sequence-262727-        1570 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  bacterial expression of GST-p18.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1570)
  AUTHORS   Shen K, Sabatini DM
  TITLE     Ragulator and SLC38A9 activate the Rag GTPases through noncanonical 
            GEF mechanisms.
  JOURNAL   Proc Natl Acad Sci U S A. 2018 Sep 18;115(38):9545-9550. doi: 
            10.1073/pnas.1811727115. Epub 2018 Sep 4.
  PUBMED    30181260
REFERENCE   2  (bases 1 to 1570)
  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: 
            "10.1073/pnas.1811727115"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2018-09-18- 18"; volume: "115"; issue: "38"; pages: 
            "9545-9550"
COMMENT     Sequence Label: [pSK122] pETDuet GST-p18(delta5)
FEATURES             Location/Qualifiers
     source          1..1570
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    7..31
                     /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)."
     RBS             46..68
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             99..752
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     710..732
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             759..776
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     1327..1346
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1327..1345
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1346..1370
                     /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)."
     CDS             1479..1523
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
ORIGIN
        1 tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa
       61 gaaggagata taccatgggc agcaacacta gtagcaatat gtcccctata ctaggttatt
      121 ggaaaattaa gggccttgtg caacccactc gacttctttt ggaatatctt gaagaaaaat
      181 atgaagagca tttgtatgag cgcgatgaag gtgataaatg gcgaaacaaa aagtttgaat
      241 tgggtttgga gtttcccaat cttccttatt atattgatgg tgatgttaaa ttaacacagt
      301 ctatggccat catacgttat atagctgaca agcacaacat gttgggtggt tgtccaaaag
      361 agcgtgcaga gatttcaatg cttgaaggag cggttttgga tattagatac ggtgtttcga
      421 gaattgcata tagtaaagac tttgaaactc tcaaagttga ttttcttagc aagctacctg
      481 aaatgctgaa aatgttcgaa gatcgtttat gtcataaaac atatttaaat ggtgatcatg
      541 taacccatcc tgacttcatg ttgtatgacg ctcttgatgt tgttttatac atggacccaa
      601 tgtgcctgga tgcgttccca aaattagttt gttttaaaaa acgtattgaa gctatcccac
      661 aaattgataa gtacttgaaa tccagcaagt atatagcatg gcctttgcag ggctggcaag
      721 ccacgtttgg tggtggcgac catcctccaa aatcggatct ggttccgcgt ggatccccga
      781 attcgagcag cgagaacgag gactcggacc aggaccgaga ggagcggaag ctgctgctgg
      841 accctagcag cccccctacc aaagctctca atggagccga gcccaactac cacagcctgc
      901 cttccgctcg cactgatgag caggccctgc tctcttccat ccttgccaag acagccagca
      961 acatcattga tgtgtctgct gcagactcac agggcatgga gcagcatgag tacatggacc
     1021 gtgccaggca gtacagcacc cgcttggctg tgctgagcag cagcctgacc cattggaaga
     1081 agctgccacc gctgccgtct cttaccagcc agccccacca agtgctggcc agtgagccca
     1141 tcccgttctc tgatttgcag caggtctcca ggatagctgc ttatgcctac agtgcacttt
     1201 ctcagatccg tgtggacgca aaagaggagc tggttgtaca gtttgggatc ccatgactcg
     1261 agcggccgca taatgcttaa gtcgaacaga aagtaatcgt attgtacacg gccgcataat
     1321 cgaaattaat acgactcact ataggggaat tgtgagcgga taacaattcc ccatcttagt
     1381 atattagtta agtataagaa ggagatatac atatggcaga tctcaattgg atatcggccg
     1441 gccacgcgat cgctgacgtc ggtaccctcg agtctggtaa agaaaccgct gctgcgaaat
     1501 ttgaacgcca gcacatggac tcgtctacta gcgcagctta attaacctag gctgctgcca
     1561 ccgctgagca
//
