LOCUS       sequence-444007-        5256 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  integrative plasmid for heterologous gene expression in S. pyogenes 
            SF370, integrates into the sagB locus via homologous recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5256)
  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 5256)
  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: pEC2796 (pSpy0C4)
FEATURES             Location/Qualifiers
     source          1..5256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(5..21)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5..21)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      2617..3205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3106..3125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3359..3376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4059..4093)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     primer_bind     4129..4145
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4255..4276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4291..4296,4297..4314,4315..4321)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4329..4345
                     /label=lac operator
                     /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)."
     RBS             4363..4374
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4381..5058
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      5103..5174
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5190..5217
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 atgtgtcata gctgtttcct gcaaagaatg gtgatgacaa ttgtacaggt tatcaataaa
       61 gccattgata ctctcattct tatcccgtat ttattaactt ctcctagtat tagtaattat
      121 catggctgtc atggcgcatt aacggaataa agggtgtgct taaatcgggc cattttgcgt
      181 aataagaaaa aggattaatt atgagcgaat tgaattaata ataaggtaat agatttacat
      241 tagaaaatga aaggggattt tatgcgtgag aatgttacag tctatcccgg caatagttac
      301 ccttattatc aagataagaa agaaaaggat ttttcgctac gctcaaatcc tttaaaaaaa
      361 cacaaaagac cacatttttt aatgtggtct tttattcttc aactaaagca cccattagtt
      421 caacaaacga aaattggata aagtgggata tttttaaaat atatatttat gttacagtaa
      481 tattgacttt taaaaaagga ttgattataa tgaagaaagc agacaagtaa gcctcctaaa
      541 ttcactttag ataaaaattt aggaggcata tcaaatgaac tttaataaaa ttcactttag
      601 ataaaaattt aggaggcata tcaaatgaac tttaataaaa ttgatttaga caattggaag
      661 agaaaagaga tatttaatca ttatttgaac caacaaacga cttttagtat aaccacagaa
      721 attgatatta gtgttttata ccgaaacata aaacaagaag gatataaatt ttaccctgca
      781 tttattttct tagtgacaag ggtgataaac tcaaatacag cttttagaac tggttacaat
      841 agcgacggag agttaggtta ttgggataag ttagagccac tttatacaat ttttgatggt
      901 gtatctaaaa cattctctgg tatttggact cctgtaaaga atgacttcaa agagttttat
      961 gatttatacc tttctgatgt agagaaatat aatggttcgg ggaaattgtt tcccaaaaca
     1021 cctatacctg aaaatgcttt ttctctttct attattccat ggacttcatt tactgggttt
     1081 aacttaaata tcaataataa tagtaattac cttctaccca ttattacagc aggaaaattc
     1141 attaataaag gtaattcaat atatttaccg ctatctttac aggtacatca ttctgtttgt
     1201 gatggttatc atgcaggatt gtttatgaac tctattcagg aattgtcaga taggcctaat
     1261 gactggcttt tataatatga gataatgccg actgtacttt ttacagtcgg ttttctaatg
     1321 tcactaacct gccccgttag ttgaagaagg tttttatatt acagctccag atctaaggag
     1381 tctcaatgaa atatcaactt aatagtaatg ttcgtgtcgt gacatttcaa gatacatttt
     1441 gttttagaaa aggaatctgg gactttaatg aagccattct agatttgaca caagaacctc
     1501 aaaatttaaa gatagtttat caagaaatag tttcacagtt ggttaaggga gtagccattg
     1561 atactgaaac ttatgaaaat actttagaac ctgagacgtt cgctaattta atggaagtca
     1621 tttcaggctt gtattacaat gatatgctga tgcttgagga tgactacaat cttgaagaaa
     1681 atgtcatgaa aattttaatg ggtaatttcc gttttatggc gcaagcaggt catatggtaa
     1741 gcaatgatcc tgttttgttt atcagtgatt cgacttatgt taatgaatct gctgagttgc
     1801 tagctgaaca actgcattta caattgcaag tggctagtga tgacttaaaa atgttaattc
     1861 agcagacaga tgttagttcg cgtttagatg ctttggagca tcgccgtaac atgaattgtt
     1921 tatcagatgc cttgtgcgat taccaaagta ttattatttg tcaagaacgc ttgaatatta
     1981 tgatgttgcg acacttgaat gaagttagcg tagctatgaa aaaacaatta gtgattggtt
     2041 ttgtagatgg accattcttg cacacttgta ctttgaatcc tccccatagt gctgactttg
     2101 atagcttaga aagaagagtt ttggcacgtt tacaagactc aactctctat caacattttg
     2161 ctaatcaggt tttaccagcc acacaagatg tgagccaggc ctatttacca ttgttaaatg
     2221 ttttaatgaa tttagtcgtt agcgaagctt tcattattgc tcaaacaggt agttctaaat
     2281 ttgagggacg tttattaagc atttacttac cgacgcttga gattcaggtc caagatattt
     2341 tgaaaatgtc taattcacag acacaaggag ctttggcaaa attaaagtat gaggatcagc
     2401 agatatcaac acgtgagatt gttaaaaaac tccttgacga gtaaccctgt cagaccaagt
     2461 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2521 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2581 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2641 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2701 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2761 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2821 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2881 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2941 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3001 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3061 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3121 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3181 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3241 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3301 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3361 cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg
     3421 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cagtggactt
     3481 cctccttggt tttaagggtt aattgttcta gtgaataaaa ggtttcttgg tcgcgcagta
     3541 gggatcaagc gagcagataa aatgcctatt ttaaatcagt tcgagcgaat tttcgtgcta
     3601 caagctcaaa agaggctact aaagtattag aatagtttat ctctgagtga agttcttgtt
     3661 ataagaagat gatgttatcg tttgaaacga ccaaaaaatc tcctaacttt tactaatttc
     3721 tacatgaaat ctagtccagt atttactcta caaccttgat tgattcgctt aataaagaca
     3781 tttctacttg attgtttaca catagttatt gatagaatct attataaaat tcagtatatt
     3841 agatcataat tgtttctata ttaatcaatt attatttttt ttgtcatttt tgataatatt
     3901 aaaaagaaag ggtttacata ttaatcattt tttactataa taaaagtgat aagaactaga
     3961 tagttgttgt gttacaacag tacaattgag ctagccttgt ccttgttgtg ttaactttat
     4021 ttttaaaata aggttaaaaa taaacgactc gcggaacgat aaaaaacgcc cggcggcaac
     4081 cgagcgttct gaattatggg gatgatgtta aggctattgg tgtttatggt aaaacgacgg
     4141 ccagtcagcg agaccagctg aattcaaaaa aaaatctaga tggccggcga ttcattaatg
     4201 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     4261 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     4321 gtgtggaatt gtgagcggat aacaatttca cacatactag agaaagagga gaaatactag
     4381 atggcttcct ccgaagacgt tatcaaagag ttcatgcgtt tcaaagttcg tatggaaggt
     4441 tccgttaacg gtcacgagtt cgaaatcgaa ggtgaaggtg aaggtcgtcc gtacgaaggt
     4501 acccagaccg ctaaactgaa agttaccaaa ggtggtccgc tgccgttcgc ttgggacatc
     4561 ctgtccccgc agttccagta cggttccaaa gcttacgtta aacacccggc tgacatcccg
     4621 gactacctga aactgtcctt cccggaaggt ttcaaatggg aacgtgttat gaacttcgaa
     4681 gacggtggtg ttgttaccgt tacccaggac tcctccctgc aagacggtga gttcatctac
     4741 aaagttaaac tgcgtggtac caacttcccg tccgatggtc cggttatgca gaaaaaaacc
     4801 atgggttggg aagcttccac cgaacgtatg tacccggaag acggtgctct gaaaggtgaa
     4861 atcaaaatgc gtctgaaact gaaagacggt ggtcactacg acgctgaagt taaaaccacc
     4921 tacatggcta aaaaaccggt tcagctgccg ggtgcttaca aaaccgacat caaactggac
     4981 atcacctccc acaacgaaga ctacaccatc gttgaacagt acgaacgtgc tgaaggtcgt
     5041 cactccaccg gtgcttaata acgctgatag tgctagtgta gatcgctact agagccaggc
     5101 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     5161 cggtgaacgc tctctactag agtcacactg gctcaccttc gggtgggcct ttctgcgttt
     5221 ataaccggtt aatactagta ctgcagtgaa ggtctc
//
