LOCUS       sequence-434502-        2933 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GST-tag fused to PreScission Protease cleavage site for protein 
            purification and cleavage.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2933)
  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 2933)
  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-848
FEATURES             Location/Qualifiers
     source          1..2933
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     39..56
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             134..787
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             794..817
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(856..874)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        942..1046
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1268..2059
                     /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      2230..2818
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2719..2738
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
       61 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatcac
      121 tctgtggtct caaatgagcc cgatcctggg ctactggaag atcaagggcc tggtgcagcc
      181 gacgcgcctg ctgctggagt acctggagga gaagtacgag gagcacctgt acgagcgcga
      241 cgagggcgac aagtggcgca acaagaagtt cgagctgggc ctggagttcc cgaacctgcc
      301 gtactacatc gacggcgacg tgaagctgac gcagagcatg gcgatcatcc gctacatcgc
      361 ggacaagcac aacatgctgg gcggctgccc gaaggagcgc gcggagatca gcatgctgga
      421 gggcgcggtg ctggacatcc gctacggcgt gagccgcatc gcgtacagca aggacttcga
      481 gacgctgaag gtggacttcc tgagcaagct gccggagatg ctgaagatgt tcgaggaccg
      541 cctgtgccac aagacgtacc tgaacggcga ccacgtgacg cacccggact tcatgctgta
      601 cgacgcgctg gacgtggtgc tgtacatgga cccgatgtgc ctggacgcgt tcccgaagct
      661 ggtgtgcttc aagaagcgca tcgaggcgat cccgcagatc gacaagtacc tgaagagcag
      721 caagtacatc gcgtggccgc tgcagggctg gcaggcgacg ttcggcggcg gcgaccaccc
      781 gccgaagagc gacctggagg tgctgttcca gggcccaggt tgagaccacg aagtggctct
      841 tcagtggacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
      901 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      961 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1021 ttcaataata ttgaaaaagg aagagtatgc gctcacgcaa ctggtccaga accttgaccg
     1081 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttt
     1141 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     1201 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     1261 aaacatcatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
     1321 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
     1381 ggatggcggc ctgaagccac acagcgatat tgatttgctg gttacggtga ccgtaaggct
     1441 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
     1501 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
     1561 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
     1621 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     1681 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     1741 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     1801 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     1861 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     1921 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     1981 cttggcctcg cgcgcagatc agttggaaga atttgtccat tacgtaaaag gcgagatcac
     2041 caaggtagtc ggcaaataac tgtcagacca agtttactca tatatacttt agattgattt
     2101 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2161 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     2221 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     2281 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     2341 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     2401 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     2461 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     2521 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     2581 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     2641 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     2701 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     2761 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     2821 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     2881 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agt
//
