LOCUS       sequence-192859-        1695 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  GST-STh expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1695)
  AUTHORS   Zhu Y, Luo Q, Davis SM, Westra C, Vickers TJ, Fleckenstein JM
  TITLE     Molecular determinants of enterotoxigenic Escherichia coli 
            heat-stable toxin secretion and delivery.
  JOURNAL   Infect Immun. 2018 Aug 20. pii: IAI.00526-18. doi: 
            10.1128/IAI.00526-18.
  PUBMED    30126899
REFERENCE   2  (bases 1 to 1695)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Infect 
            Immun. 2018 Aug 20. pii: IAI.00526-18. doi: 10.1128/IAI.00526-18."
COMMENT     Sequence Label: pCW002
FEATURES             Location/Qualifiers
     source          1..1695
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        join(10..15,16..31,32..38)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    46..62
                     /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     51..73
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             85..738
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     696..718
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             745..762
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(924..946)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1091..1109)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1177..1281
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1500..1519)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aatgagctgt tgacaattaa tcatcggctc gtataatgtg tggaattgtg agcggataac
       61 aatttcacac aggaaacagt attcatgtcc cctatactag gttattggaa aattaagggc
      121 cttgtgcaac ccactcgact tcttttggaa tatcttgaag aaaaatatga agagcatttg
      181 tatgagcgcg atgaaggtga taaatggcga aacaaaaagt ttgaattggg tttggagttt
      241 cccaatcttc cttattatat tgatggtgat gttaaattaa cacagtctat ggccatcata
      301 cgttatatag ctgacaagca caacatgttg ggtggttgtc caaaagagcg tgcagagatt
      361 tcaatgcttg aaggagcggt tttggatatt agatacggtg tttcgagaat tgcatatagt
      421 aaagactttg aaactctcaa agttgatttt cttagcaagc tacctgaaat gctgaaaatg
      481 ttcgaagatc gtttatgtca taaaacatat ttaaatggtg atcatgtaac ccatcctgac
      541 ttcatgttgt atgacgctct tgatgttgtt ttatacatgg acccaatgtg cctggatgcg
      601 ttcccaaaat tagtttgttt taaaaaacgt attgaagcta tcccacaaat tgataagtac
      661 ttgaaatcca gcaagtatat agcatggcct ttgcagggct ggcaagccac gtttggtggt
      721 ggcgaccatc ctccaaaatc ggatctggtt ccgcgtggat ccgatccccg ggtaccgagc
      781 tcgaatagta gcaattactg ctgtgaattg tgttgtaatc ctgcttgtac cgggtgctat
      841 tgagaattcc cgggtcgact cgagcggccg catcgtgact gactgacgat ctgcctcgcg
      901 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
      961 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     1021 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga
     1081 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     1141 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     1201 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1261 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1321 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1381 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1441 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1501 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     1561 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1621 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1681 aacttacttc tgaca
//
