LOCUS       sequence-434515-        3017 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mCerulean fluorescent protein fused to Factor Xa Protease cleavage 
            site and Strep-Tag II.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3017)
  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 3017)
  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-859
FEATURES             Location/Qualifiers
     source          1..3017
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..156
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             237..248
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             963..974
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             975..998
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     primer_bind     complement(1040..1058)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1126..1230
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1452..2243
                     /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      2414..3002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2903..2922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
       61 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
      121 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
      181 aaaccgcctc tccccgcgcg ttggccgatt cattaatcac tctgtggtct caaatgatcg
      241 agggcagggt gagcaagggc gaggagctgt tcaccggcgt ggtgcccatc ctggtggagc
      301 tggacggcga cgtgaacggc cacaagttca gcgtgagcgg cgagggcgag ggcgacgcca
      361 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      421 ccaccctggt gaccaccctg agctggggcg tgcagtgctt cgcccgctac cccgaccaca
      481 tgaagcagca cgacttcttc aagagcgcca tgcccgaggg ctacgtgcag gagcgcacca
      541 tcttcttcaa ggacgacggt aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      601 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      661 gccacaagct ggagtacaac gccatccacg gcaacgtgta catcaccgcc gacaagcaga
      721 agaacggcat caaggccaac ttcggcctga actgcaacat cgaggacggc agcgtgcagc
      781 tggccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
      841 accactacct gagcacccag agcaagctga gcaaggaccc caacgagaag cgcgaccaca
      901 tggtgctgct ggagttcgtg accgccgccg gcatcaccct gggcatggac gagctgtaca
      961 agatcgaggg caggtggagc cacccgcagt tcgagaaggg ttcgtgagac cacgaagtgg
     1021 ctcttcagtg gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     1081 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     1141 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     1201 atgcttcaat aatattgaaa aaggaagagt atgcgctcac gcaactggtc cagaaccttg
     1261 accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta tgactgtttt
     1321 tttttggggt acagtctatg cctcgggcat ccaagcagca agcgcgttac gccgtgggtc
     1381 gatgtttgat gttatggagc agcaacgatg ttacgcagca gggcagtcgc cctaaaacaa
     1441 agttaaacat catgagggaa gcggtgatcg ccgaagtatc gactcaacta tcagaggtag
     1501 ttggcgtcat cgagcgccat ctcgaaccga cgttgctggc cgtacatttg tacggctccg
     1561 cagtggatgg cggcctgaag ccacacagcg atattgattt gctggttacg gtgaccgtaa
     1621 ggcttgatga aacaacgcgg cgagctttga tcaacgacct tttggaaact tcggcttccc
     1681 ctggagagag cgagattctc cgcgctgtag aagtcaccat tgttgtgcac gacgacatca
     1741 ttccgtggcg ttatccagct aagcgcgaac tgcaatttgg agaatggcag cgcaatgaca
     1801 ttcttgcagg tatcttcgag ccagccacga tcgacattga tctggctatc ttgctgacaa
     1861 aagcaagaga acatagcgtt gccttggtag gtccagcggc ggaggaactc tttgatccgg
     1921 ttcctgaaca ggatctattt gaggcgctaa atgaaacctt aacgctatgg aactcgccgc
     1981 ccgactgggc tggcgatgag cgaaatgtag tgcttacgtt gtcccgcatt tggtacagcg
     2041 cagtaaccgg caaaatcgcg ccgaaggatg tcgctgccga ctgggcaatg gagcgcctgc
     2101 cggcccagta tcagcccgtc atacttgaag ctagacaggc ttatcttgga caagaagaag
     2161 atcgcttggc ctcgcgcgca gatcagttgg aagaatttgt ccattacgta aaaggcgaga
     2221 tcaccaaggt agtcggcaaa taactgtcag accaagttta ctcatatata ctttagattg
     2281 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2341 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2401 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2461 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2521 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2581 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     2641 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     2701 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     2761 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     2821 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     2881 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     2941 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3001 aaaacgccag caacgcg
//
