LOCUS       sequence-434859-        2937 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST-tag fused to TEV Protease cleavage site for protein purification
            and cleavage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2937)
  AUTHORS   Hieronimus K, Donauer T, Klein J, Hinkel B, Spanle JV, Probst A, 
            Niemeyer J, Kibrom S, Kiefer AM, Schneider L, Husemann B, Bischoff 
            E, Mohring S, Bayer N, Klein D, Engels A, Ziehmer BG, Stiebeta J, 
            Moroka P, Schroda M, Deponte M
  TITLE     A Modular Cloning Toolkit for the production of recombinant proteins
            in Leishmania tarentolae.
  JOURNAL   Microb Cell. 2024 Apr 30;11:128-142. doi: 10.15698/mic2024.04.821. 
            eCollection 2024.
  PUBMED    38799406
REFERENCE   2  (bases 1 to 2937)
  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: "Microb 
            Cell."; date: "2024-04-30"; volume: "11:128-142. doi"; pages: " 
            10.15698/mic2024.04.821. eCollection 2024"
COMMENT     Sequence Label: pMBS-864
FEATURES             Location/Qualifiers
     source          1..2937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     386..405
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     639..656
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             737..757
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             758..1411
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     complement(1460..1478)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1546..1650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1872..2663
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      join(2834..2937,1..485)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
       61 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      121 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      181 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      241 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      301 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      361 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      421 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      481 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
      541 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
      601 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
      661 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatcac
      721 tctgtggtct cattcggaga acctgtactt ccagagcatg agcccgatcc tgggctactg
      781 gaagatcaag ggcctggtgc agccgacgcg cctgctgctg gagtacctgg aggagaagta
      841 cgaggagcac ctgtacgagc gcgacgaggg cgacaagtgg cgcaacaaga agttcgagct
      901 gggcctggag ttcccgaacc tgccgtacta catcgacggc gacgtgaagc tgacgcagag
      961 catggcgatc atccgctaca tcgcggacaa gcacaacatg ctgggcggct gcccgaagga
     1021 gcgcgcggag atcagcatgc tggagggcgc ggtgctggac atccgctacg gcgtgagccg
     1081 catcgcgtac agcaaggact tcgagacgct gaaggtggac ttcctgagca agctgccgga
     1141 gatgctgaag atgttcgagg accgcctgtg ccacaagacg tacctgaacg gcgaccacgt
     1201 gacgcacccg gacttcatgc tgtacgacgc gctggacgtg gtgctgtaca tggacccgat
     1261 gtgcctggac gcgttcccga agctggtgtg cttcaagaag cgcatcgagg cgatcccgca
     1321 gatcgacaag tacctgaaga gcagcaagta catcgcgtgg ccgctgcagg gctggcaggc
     1381 gacgttcggc ggcggcgacc acccgccgaa gagcgactaa gctttgagac cacgaagtgg
     1441 ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1501 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1621 atgcttcaat aatattgaaa aaggaagagt atgcgctcac gcaactggtc cagaaccttg
     1681 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     1741 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     1801 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     1861 agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     1921 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     1981 cagtggatgg cggcctgaag ccacacagcg atattgattt gctggttacg gtgaccgtaa
     2041 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     2101 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     2161 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     2221 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     2281 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     2341 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     2401 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     2461 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     2521 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     2581 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccattacgta aaaggcgaga
     2641 tcaccaaggt agtcggcaaa taactgtcag accaagttta ctcatatata ctttagattg
     2701 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2761 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2821 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2881 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttc
//
