LOCUS       sequence-444093-        5611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  replicative E. coli-S. pyogenes shuttle plasmid for zinc-inducible 
            expression of the Firefly luciferase in S. pyogenes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5611)
  AUTHORS   Lautenschlager N, Schmidt K, Schiffer C, Wulff TF, Hahnke K, 
            Finstermeier K, Mansour M, Elsholz AKW, Charpentier E
  TITLE     Expanding the genetic toolbox for the obligate human pathogen 
            Streptococcus pyogenes.
  JOURNAL   Front Bioeng Biotechnol. 2024 Jun 7;12:1395659. doi: 
            10.3389/fbioe.2024.1395659. eCollection 2024.
  PUBMED    38911550
REFERENCE   2  (bases 1 to 5611)
  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: "Front 
            Bioeng Biotechnol."; date: "2024-06-7"; volume: "12:1395659. doi"; 
            pages: " 10.3389/fbioe.2024.1395659. eCollection 2024"
COMMENT     Sequence Label: pEC3006 (pSpy1C-PZn_ffluc)
FEATURES             Location/Qualifiers
     source          1..5611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..18)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2..18)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2954..2965
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(3720..3754)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     3790..3806
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3943..5595
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(4009..4027)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 tgtcatagct gtttcctgca aagaatggtg atgacaattg tacaggttat caataaagcc
       61 attgatactc tcattcttat cccgtattta ttaacttctc ctagtattag taattatcat
      121 ggctgtcatg gcgcattaac ggaataaagg gtgtgcttaa atcgggccat tttgcgtaat
      181 aagaaaaagg attaattatg agcgaattga attaataata aggtaataga tttacattag
      241 aaaatgaaag gggattttat gcgtgagaat gttacagtct atcccggcaa tagttaccct
      301 tattatcaag ataagaaaga aaaggatttt tcgctacgct caaatccttt aaaaaaacac
      361 aaaagaccac attttttaat gtggtctttt attcttcaac taaagcaccc attagttcaa
      421 caaacgaaaa ttggataaag tgggatattt ttaaaatata tatttatgtt acagtaatat
      481 tgacttttaa aaaaggattg attataatga agaaagcaga caagtaagcc tcctaaattc
      541 actttagata aaaatttagg aggcatatca aatgaacttt aataaaattc actttagata
      601 aaaatttagg aggcatatca aatgaacttt aataaaattg atttagacaa ttggaagaga
      661 aaagagatat ttaatcatta tttgaaccaa caaacgactt ttagtataac cacagaaatt
      721 gatattagtg ttttataccg aaacataaaa caagaaggat ataaatttta ccctgcattt
      781 attttcttag tgacaagggt gataaactca aatacagctt ttagaactgg ttacaatagc
      841 gacggagagt taggttattg ggataagtta gagccacttt atacaatttt tgatggtgta
      901 tctaaaacat tctctggtat ttggactcct gtaaagaatg acttcaaaga gttttatgat
      961 ttataccttt ctgatgtaga gaaatataat ggttcgggga aattgtttcc caaaacacct
     1021 atacctgaaa atgctttttc tctttctatt attccatgga cttcatttac tgggtttaac
     1081 ttaaatatca ataataatag taattacctt ctacccatta ttacagcagg aaaattcatt
     1141 aataaaggta attcaatata tttaccgcta tctttacagg tacatcattc tgtttgtgat
     1201 ggttatcatg caggattgtt tatgaactct attcaggaat tgtcagatag gcctaatgac
     1261 tggcttttat aatatgagat aatgccgact gtacttttta cagtcggttt tctaatgtca
     1321 ctaacctgcc ccgttagttg aagaaggttt ttatattaca gctccagatc catatccttc
     1381 tttttctgaa ccgacttctc ctttttcgct tctttattcc aattgcttta ttgacgttga
     1441 gcctcggaac ccttaacaat cccaaaactt gtcgactttc gtcagggggg cttttattta
     1501 ttcaataatc cctcctctca ataaatctat tgttgtactt aattcaactt ccatttctct
     1561 gtatctttca atacgctctt ttaagtcctt aatttctttt tttaattcct cattttcagc
     1621 aaataactct ttttctttgt ttgtcatttt atttcccccg tttcagcatc aagaaccttt
     1681 gcataacttg ctctatatcc acactgataa ttgccctcaa accataatct aaaggcgcta
     1741 gagtttgttg aaacaatatc ttttacatca ttcgtattta aaattccaaa ctccgctccc
     1801 ctaaggcgaa taaaagccat taactctttt gtatttacca aattatagtc atccactata
     1861 tctaaaagta aattcttcaa ttttcttttt tggctttcat caagtgttat atagcggtca
     1921 atatcaaaat cattaatgtt caaaatatct tttttgtcgt atatatgttt attcttagca
     1981 atagcgtcct ttgattcatg agtcaaatat tcatatgaac ctttgatata atcaagtatc
     2041 tcaacatgag caactgaact attccccaat tttcgcttaa tcttgttcct aacgctttct
     2101 attgttacag gatttcgtgc aatatatata acgtgatagt gtggtttttt atagtgcttt
     2161 ccatttcgta taacatcact actattccat gtatctttat cttttttttc gtccatatcg
     2221 tgtaaaggac tgacagccat agatacgccc aaactctcta atttttcttt ccaatcatta
     2281 ggaattgagt caggatataa taaaaatcca aaatttctag ctttagtatt tttaatagcc
     2341 atgatataat taccttatca aaaacaagta gcgaaaactc gtatccttct aaaaacgcga
     2401 gctttcgctt attttttttg ttctgattcc tttcttgcat attcttctat agctaacgcc
     2461 gcaaccgcag attttgaaaa acctttttgt ttcgccatat ctgttaattt tttatcttgc
     2521 tcttttgtca gagaaatcat aactcttttt ttcgattctg aaatcaccat ttaaaaaact
     2581 ccaatcaaat aattttataa aattagtgta tcactttgta atcataaaaa caacaataaa
     2641 gctacttaaa tatagattta taaaaaacgt tggcgaaaac gttggcgatt cgttggcgat
     2701 tgaaaaaccc ctcaaaccct tgagccagtt gggatagagc gtttttggca caaaaattgg
     2761 cactcggcac ttaatggggg gtcgtagtac ggaagcaaaa ttcgcttcct ttccccccat
     2821 ttttttccaa attccaaatt tttttcaaaa attttccagc gctaccgctc ggcaaaattg
     2881 caagcaattt ttaaaatcaa acccatgagg gaatttcatt ccctcaaact cccttgagcc
     2941 tcctccaacc gaaatagaag gacgctgcgc ttattatttc attcagtcat cggctttcat
     3001 aatctaacag acaacatctt cgctgcaaag ccacgctacg ctcaagggct tttacgctac
     3061 gataacgcct gttttaacga ttatgccgat aactaaacga aataaacgct aaaacgtcac
     3121 agaaacgatt ttcagacgtt ttaataaaaa atcgaccgaa aaataaatat aaatcggata
     3181 tacaatcggc aattgacgaa actgcaaaat atcctgtaaa ggatacggat tttatgaccg
     3241 atgatgaaga aaagaatttg aaacgtttgt ctgatttgga ggaaggttta caccgtaaaa
     3301 ggttaatctc ctatggtggt ttgttaaaag aaatacataa aaaattaaac cttgatgaca
     3361 cagaagaagg cgatttgatt catacagatg atgacgaaaa agccgatgaa gatggatttt
     3421 ctattattgc aatgtggaat tgggaacgga aaaattattt tattaaagag tagttcaaca
     3481 aacgggccag tttgttgaag attagatgct ataattgtta ttaaaaggat tgaaggatgc
     3541 ttaggaagac gagttattaa tagctgaata agaacggtgc tctccaaata ttcttattta
     3601 gaaaagcaaa tctaaaatta tctgaaaagg gaatgagaat agtgaatgga ccaataataa
     3661 tgactagaga agaaagaatg aagattgttc atgaaattaa ggaacgaata ttggataaca
     3721 taaaaaacgc ccggcggcaa ccgagcgttc tgaattatgg ggatgatgtt aaggctattg
     3781 gtgtttatgg taaaacgacg gccagtcatc ggacactttc taaatgttgt tcactattga
     3841 acaggtgctc atttttggtt atagtaattg tattttaact tttagataat agaataaagc
     3901 attctactat acatattacc agtataacaa aaggagaaaa ttatggaaga cgccaaaaac
     3961 ataaagaaag gcccggcgcc attctatcct ctagaggatg gaaccgctgg agagcaactg
     4021 cataaggcta tgaagagata cgccctggtt cctggaacaa ttgcttttac agatgcacat
     4081 atcgaggtga acatcacgta cgcggaatac ttcgaaatgt ccgttcggtt ggcagaagct
     4141 atgaaacgat atgggctgaa tacaaatcac agaatcgtcg tatgcagtga aaactctctt
     4201 caattcttta tgccggtgtt gggcgcgtta tttatcggag ttgcagttgc gcccgcgaac
     4261 gacatttata atgaacgtga attgctcaac agtatgaaca tttcgcagcc taccgtagtg
     4321 tttgtttcca aaaaggggtt gcaaaaaatt ttgaacgtgc aaaaaaaatt accaataatc
     4381 cagaaaatta ttatcatgga ttctaaaacg gattaccagg gatttcagtc gatgtacacg
     4441 ttcgtcacat ctcatctacc tcccggtttt aatgaatacg attttgtacc agagtccttt
     4501 gatcgtgaca aaacaattgc actgataatg aattcctctg gatctactgg gttacctaag
     4561 ggtgtggccc ttccgcatag aactgcctgc gtcagattct cgcatgccag agatcctatt
     4621 tttggcaatc aaatcattcc ggatactgcg attttaagtg ttgttccatt ccatcacggt
     4681 tttggaatgt ttactacact cggatatttg atatgtggat ttcgagtcgt cttaatgtat
     4741 agatttgaag aagagctgtt tttacgatcc cttcaggatt acaaaattca aagtgcgttg
     4801 ctagtaccaa ccctattttc attcttcgcc aaaagcactc tgattgacaa atacgattta
     4861 tctaatttac acgaaattgc ttctgggggc gcacctcttt cgaaagaagt cggggaagcg
     4921 gttgcaaaac gcttccatct tccagggata cgacaaggat atgggctcac tgagactaca
     4981 tcagctattc tgattacacc cgagggggat gataaaccgg gcgcggtcgg taaagttgtt
     5041 ccattttttg aagcgaaggt tgtggatctg gataccggga aaacgctggg cgttaatcag
     5101 agaggcgaat tatgtgtcag aggacctatg attatgtccg gttatgtaaa caatccggaa
     5161 gcgaccaacg ccttgattga caaggatgga tggctacatt ctggagacat agcttactgg
     5221 gacgaagacg aacacttctt catagttgac cgcttgaagt ctttaattaa atacaaagga
     5281 tatcaggtgg cccccgctga attggaatcg atattgttac aacaccccaa catcttcgac
     5341 gcgggcgtgg caggtcttcc cgacgatgac gccggtgaac ttcccgccgc cgttgttgtt
     5401 ttggagcacg gaaagacgat gacggaaaaa gagatcgtgg attacgtcgc cagtcaagta
     5461 acaaccgcga aaaagttgcg cggaggagtt gtgtttgtgg acgaagtacc gaaaggtctt
     5521 accggaaaac tcgacgcaag aaaaatcaga gagatcctca taaaggccaa gaagggcgga
     5581 aagtccaaat tgtaatacta gtactgcagt g
//
