LOCUS       sequence-434498-        2360 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  3xFLAG-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 2360)
  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 2360)
  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-846
FEATURES             Location/Qualifiers
     source          1..2360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             37..108
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags"
                     /label=3xFLAG
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             118..141
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(183..201)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        269..373
                     /gene="bla"
                     /label=AmpR promoter
     CDS             595..1386
                     /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      1557..2145
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2046..2065
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2299..2316
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ttggccgatt cattaatcac tctgtggtct caaatggact acaaagacga tgatgataag
       61 gattacaagg acgacgatga caaggactac aaggatgatg atgacaagcg cggcagcctg
      121 gaggtgctgt tccagggccc ggcaggttga gaccacgaag tggctcttca gtggacgaaa
      181 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
      241 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
      301 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
      361 aaaaaggaag agtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg
      421 taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttttgg ggtacagtct
      481 atgcctcggg catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg
      541 agcagcaacg atgttacgca gcagggcagt cgccctaaaa caaagttaaa catcatgagg
      601 gaagcggtga tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc
      661 catctcgaac cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg
      721 aagccacaca gcgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg
      781 cggcgagctt tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt
      841 ctccgcgctg tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca
      901 gctaagcgcg aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc
      961 gagccagcca cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc
     1021 gttgccttgg taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta
     1081 tttgaggcgc taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat
     1141 gagcgaaatg tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc
     1201 gcgccgaagg atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc
     1261 gtcatacttg aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc
     1321 gcagatcagt tggaagaatt tgtccattac gtaaaaggcg agatcaccaa ggtagtcggc
     1381 aaataactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     1441 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     1501 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1561 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1621 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1681 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1741 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1801 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1861 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1921 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1981 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     2041 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     2101 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     2161 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     2221 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     2281 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2341 aaaccgcctc tccccgcgcg
//
