LOCUS       sequence-444100-        5638 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  replicative E. coli-S. pyogenes shuttle plasmid for IPTG-inducible 
            expression of the Firefly luciferase in a strain expressing lacI 
            repressor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5638)
  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 5638)
  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: pEC3109 (pSpy1C-Plac(1)_ffluc)
FEATURES             Location/Qualifiers
     source          1..5638
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(4..20)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4..20)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2956..2967
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      complement(3722..3756)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     3792..3808
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3905..3924
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             3972..5624
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(4038..4056)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
ORIGIN
        1 tgtgtcatag ctgtttcctg caaagaatgg tgatgacaat tgtacaggtt atcaataaag
       61 ccattgatac tctcattctt atcccgtatt tattaacttc tcctagtatt agtaattatc
      121 atggctgtca tggcgcatta acggaataaa gggtgtgctt aaatcgggcc attttgcgta
      181 ataagaaaaa ggattaatta tgagcgaatt gaattaataa taaggtaata gatttacatt
      241 agaaaatgaa aggggatttt atgcgtgaga atgttacagt ctatcccggc aatagttacc
      301 cttattatca agataagaaa gaaaaggatt tttcgctacg ctcaaatcct ttaaaaaaac
      361 acaaaagacc acatttttta atgtggtctt ttattcttca actaaagcac ccattagttc
      421 aacaaacgaa aattggataa agtgggatat ttttaaaata tatatttatg ttacagtaat
      481 attgactttt aaaaaaggat tgattataat gaagaaagca gacaagtaag cctcctaaat
      541 tcactttaga taaaaattta ggaggcatat caaatgaact ttaataaaat tcactttaga
      601 taaaaattta ggaggcatat caaatgaact ttaataaaat tgatttagac aattggaaga
      661 gaaaagagat atttaatcat tatttgaacc aacaaacgac ttttagtata accacagaaa
      721 ttgatattag tgttttatac cgaaacataa aacaagaagg atataaattt taccctgcat
      781 ttattttctt agtgacaagg gtgataaact caaatacagc ttttagaact ggttacaata
      841 gcgacggaga gttaggttat tgggataagt tagagccact ttatacaatt tttgatggtg
      901 tatctaaaac attctctggt atttggactc ctgtaaagaa tgacttcaaa gagttttatg
      961 atttatacct ttctgatgta gagaaatata atggttcggg gaaattgttt cccaaaacac
     1021 ctatacctga aaatgctttt tctctttcta ttattccatg gacttcattt actgggttta
     1081 acttaaatat caataataat agtaattacc ttctacccat tattacagca ggaaaattca
     1141 ttaataaagg taattcaata tatttaccgc tatctttaca ggtacatcat tctgtttgtg
     1201 atggttatca tgcaggattg tttatgaact ctattcagga attgtcagat aggcctaatg
     1261 actggctttt ataatatgag ataatgccga ctgtactttt tacagtcggt tttctaatgt
     1321 cactaacctg ccccgttagt tgaagaaggt ttttatatta cagctccaga tccatatcct
     1381 tctttttctg aaccgacttc tcctttttcg cttctttatt ccaattgctt tattgacgtt
     1441 gagcctcgga acccttaaca atcccaaaac ttgtcgactt tcgtcagggg ggcttttatt
     1501 tattcaataa tccctcctct caataaatct attgttgtac ttaattcaac ttccatttct
     1561 ctgtatcttt caatacgctc ttttaagtcc ttaatttctt tttttaattc ctcattttca
     1621 gcaaataact ctttttcttt gtttgtcatt ttatttcccc cgtttcagca tcaagaacct
     1681 ttgcataact tgctctatat ccacactgat aattgccctc aaaccataat ctaaaggcgc
     1741 tagagtttgt tgaaacaata tcttttacat cattcgtatt taaaattcca aactccgctc
     1801 ccctaaggcg aataaaagcc attaactctt ttgtatttac caaattatag tcatccacta
     1861 tatctaaaag taaattcttc aattttcttt tttggctttc atcaagtgtt atatagcggt
     1921 caatatcaaa atcattaatg ttcaaaatat cttttttgtc gtatatatgt ttattcttag
     1981 caatagcgtc ctttgattca tgagtcaaat attcatatga acctttgata taatcaagta
     2041 tctcaacatg agcaactgaa ctattcccca attttcgctt aatcttgttc ctaacgcttt
     2101 ctattgttac aggatttcgt gcaatatata taacgtgata gtgtggtttt ttatagtgct
     2161 ttccatttcg tataacatca ctactattcc atgtatcttt atcttttttt tcgtccatat
     2221 cgtgtaaagg actgacagcc atagatacgc ccaaactctc taatttttct ttccaatcat
     2281 taggaattga gtcaggatat aataaaaatc caaaatttct agctttagta tttttaatag
     2341 ccatgatata attaccttat caaaaacaag tagcgaaaac tcgtatcctt ctaaaaacgc
     2401 gagctttcgc ttattttttt tgttctgatt cctttcttgc atattcttct atagctaacg
     2461 ccgcaaccgc agattttgaa aaaccttttt gtttcgccat atctgttaat tttttatctt
     2521 gctcttttgt cagagaaatc ataactcttt ttttcgattc tgaaatcacc atttaaaaaa
     2581 ctccaatcaa ataattttat aaaattagtg tatcactttg taatcataaa aacaacaata
     2641 aagctactta aatatagatt tataaaaaac gttggcgaaa acgttggcga ttcgttggcg
     2701 attgaaaaac ccctcaaacc cttgagccag ttgggataga gcgtttttgg cacaaaaatt
     2761 ggcactcggc acttaatggg gggtcgtagt acggaagcaa aattcgcttc ctttcccccc
     2821 atttttttcc aaattccaaa tttttttcaa aaattttcca gcgctaccgc tcggcaaaat
     2881 tgcaagcaat ttttaaaatc aaacccatga gggaatttca ttccctcaaa ctcccttgag
     2941 cctcctccaa ccgaaataga aggacgctgc gcttattatt tcattcagtc atcggctttc
     3001 ataatctaac agacaacatc ttcgctgcaa agccacgcta cgctcaaggg cttttacgct
     3061 acgataacgc ctgttttaac gattatgccg ataactaaac gaaataaacg ctaaaacgtc
     3121 acagaaacga ttttcagacg ttttaataaa aaatcgaccg aaaaataaat ataaatcgga
     3181 tatacaatcg gcaattgacg aaactgcaaa atatcctgta aaggatacgg attttatgac
     3241 cgatgatgaa gaaaagaatt tgaaacgttt gtctgatttg gaggaaggtt tacaccgtaa
     3301 aaggttaatc tcctatggtg gtttgttaaa agaaatacat aaaaaattaa accttgatga
     3361 cacagaagaa ggcgatttga ttcatacaga tgatgacgaa aaagccgatg aagatggatt
     3421 ttctattatt gcaatgtgga attgggaacg gaaaaattat tttattaaag agtagttcaa
     3481 caaacgggcc agtttgttga agattagatg ctataattgt tattaaaagg attgaaggat
     3541 gcttaggaag acgagttatt aatagctgaa taagaacggt gctctccaaa tattcttatt
     3601 tagaaaagca aatctaaaat tatctgaaaa gggaatgaga atagtgaatg gaccaataat
     3661 aatgactaga gaagaaagaa tgaagattgt tcatgaaatt aaggaacgaa tattggataa
     3721 cataaaaaac gcccggcggc aaccgagcgt tctgaattat ggggatgatg ttaaggctat
     3781 tggtgtttat ggtaaaacga cggccagtct tatagaattc actagtaatt gtgaggcgga
     3841 taacaattct cactcattct acagtttatt cttgacattg cactgtcccc ctggtataat
     3901 aactaattgt gagcgctcac aattaagatc cccggggcgg ccgcgaattc aaaagatcta
     3961 aggaggaaca tatggaagac gccaaaaaca taaagaaagg cccggcgcca ttctatcctc
     4021 tagaggatgg aaccgctgga gagcaactgc ataaggctat gaagagatac gccctggttc
     4081 ctggaacaat tgcttttaca gatgcacata tcgaggtgaa catcacgtac gcggaatact
     4141 tcgaaatgtc cgttcggttg gcagaagcta tgaaacgata tgggctgaat acaaatcaca
     4201 gaatcgtcgt atgcagtgaa aactctcttc aattctttat gccggtgttg ggcgcgttat
     4261 ttatcggagt tgcagttgcg cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca
     4321 gtatgaacat ttcgcagcct accgtagtgt ttgtttccaa aaaggggttg caaaaaattt
     4381 tgaacgtgca aaaaaaatta ccaataatcc agaaaattat tatcatggat tctaaaacgg
     4441 attaccaggg atttcagtcg atgtacacgt tcgtcacatc tcatctacct cccggtttta
     4501 atgaatacga ttttgtacca gagtcctttg atcgtgacaa aacaattgca ctgataatga
     4561 attcctctgg atctactggg ttacctaagg gtgtggccct tccgcataga actgcctgcg
     4621 tcagattctc gcatgccaga gatcctattt ttggcaatca aatcattccg gatactgcga
     4681 ttttaagtgt tgttccattc catcacggtt ttggaatgtt tactacactc ggatatttga
     4741 tatgtggatt tcgagtcgtc ttaatgtata gatttgaaga agagctgttt ttacgatccc
     4801 ttcaggatta caaaattcaa agtgcgttgc tagtaccaac cctattttca ttcttcgcca
     4861 aaagcactct gattgacaaa tacgatttat ctaatttaca cgaaattgct tctgggggcg
     4921 cacctctttc gaaagaagtc ggggaagcgg ttgcaaaacg cttccatctt ccagggatac
     4981 gacaaggata tgggctcact gagactacat cagctattct gattacaccc gagggggatg
     5041 ataaaccggg cgcggtcggt aaagttgttc cattttttga agcgaaggtt gtggatctgg
     5101 ataccgggaa aacgctgggc gttaatcaga gaggcgaatt atgtgtcaga ggacctatga
     5161 ttatgtccgg ttatgtaaac aatccggaag cgaccaacgc cttgattgac aaggatggat
     5221 ggctacattc tggagacata gcttactggg acgaagacga acacttcttc atagttgacc
     5281 gcttgaagtc tttaattaaa tacaaaggat atcaggtggc ccccgctgaa ttggaatcga
     5341 tattgttaca acaccccaac atcttcgacg cgggcgtggc aggtcttccc gacgatgacg
     5401 ccggtgaact tcccgccgcc gttgttgttt tggagcacgg aaagacgatg acggaaaaag
     5461 agatcgtgga ttacgtcgcc agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg
     5521 tgtttgtgga cgaagtaccg aaaggtctta ccggaaaact cgacgcaaga aaaatcagag
     5581 agatcctcat aaaggccaag aagggcggaa agtccaaatt gtaatactag tactgcag
//
