LOCUS       sequence-444097-        4919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  replicative E. coli-S. pyogenes shuttle plasmid for heterologous 
            gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4919)
  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 4919)
  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: pEC2483 (pSpy1C)
FEATURES             Location/Qualifiers
     source          1..4919
                     /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"
     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    3918..3939
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3954..3959,3960..3977,3978..3984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3992..4008
                     /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             4026..4037
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             4044..4721
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      4766..4837
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4853..4880
                     /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 atccatatcc
     1381 ttctttttct gaaccgactt ctcctttttc gcttctttat tccaattgct ttattgacgt
     1441 tgagcctcgg aacccttaac aatcccaaaa cttgtcgact ttcgtcaggg gggcttttat
     1501 ttattcaata atccctcctc tcaataaatc tattgttgta cttaattcaa cttccatttc
     1561 tctgtatctt tcaatacgct cttttaagtc cttaatttct ttttttaatt cctcattttc
     1621 agcaaataac tctttttctt tgtttgtcat tttatttccc ccgtttcagc atcaagaacc
     1681 tttgcataac ttgctctata tccacactga taattgccct caaaccataa tctaaaggcg
     1741 ctagagtttg ttgaaacaat atcttttaca tcattcgtat ttaaaattcc aaactccgct
     1801 cccctaaggc gaataaaagc cattaactct tttgtattta ccaaattata gtcatccact
     1861 atatctaaaa gtaaattctt caattttctt ttttggcttt catcaagtgt tatatagcgg
     1921 tcaatatcaa aatcattaat gttcaaaata tcttttttgt cgtatatatg tttattctta
     1981 gcaatagcgt cctttgattc atgagtcaaa tattcatatg aacctttgat ataatcaagt
     2041 atctcaacat gagcaactga actattcccc aattttcgct taatcttgtt cctaacgctt
     2101 tctattgtta caggatttcg tgcaatatat ataacgtgat agtgtggttt tttatagtgc
     2161 tttccatttc gtataacatc actactattc catgtatctt tatctttttt ttcgtccata
     2221 tcgtgtaaag gactgacagc catagatacg cccaaactct ctaatttttc tttccaatca
     2281 ttaggaattg agtcaggata taataaaaat ccaaaatttc tagctttagt atttttaata
     2341 gccatgatat aattacctta tcaaaaacaa gtagcgaaaa ctcgtatcct tctaaaaacg
     2401 cgagctttcg cttatttttt ttgttctgat tcctttcttg catattcttc tatagctaac
     2461 gccgcaaccg cagattttga aaaacctttt tgtttcgcca tatctgttaa ttttttatct
     2521 tgctcttttg tcagagaaat cataactctt tttttcgatt ctgaaatcac catttaaaaa
     2581 actccaatca aataatttta taaaattagt gtatcacttt gtaatcataa aaacaacaat
     2641 aaagctactt aaatatagat ttataaaaaa cgttggcgaa aacgttggcg attcgttggc
     2701 gattgaaaac 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 cggccagtca gcgagaccag ctgaattcaa aaaaaaatct
     3841 agatggccgg cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     3901 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     3961 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacatac
     4021 tagagaaaga ggagaaatac tagatggctt cctccgaaga cgttatcaaa gagttcatgc
     4081 gtttcaaagt tcgtatggaa ggttccgtta acggtcacga gttcgaaatc gaaggtgaag
     4141 gtgaaggtcg tccgtacgaa ggtacccaga ccgctaaact gaaagttacc aaaggtggtc
     4201 cgctgccgtt cgcttgggac atcctgtccc cgcagttcca gtacggttcc aaagcttacg
     4261 ttaaacaccc ggctgacatc ccggactacc tgaaactgtc cttcccggaa ggtttcaaat
     4321 gggaacgtgt tatgaacttc gaagacggtg gtgttgttac cgttacccag gactcctccc
     4381 tgcaagacgg tgagttcatc tacaaagtta aactgcgtgg taccaacttc ccgtccgatg
     4441 gtccggttat gcagaaaaaa accatgggtt gggaagcttc caccgaacgt atgtacccgg
     4501 aagacggtgc tctgaaaggt gaaatcaaaa tgcgtctgaa actgaaagac ggtggtcact
     4561 acgacgctga agttaaaacc acctacatgg ctaaaaaacc ggttcagctg ccgggtgctt
     4621 acaaaaccga catcaaactg gacatcacct cccacaacga agactacacc atcgttgaac
     4681 agtacgaacg tgctgaaggt cgtcactcca ccggtgctta ataacgctga tagtgctagt
     4741 gtagatcgct actagagcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc
     4801 ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca ctggctcacc
     4861 ttcgggtggg cctttctgcg tttataaccg gttaatacta gtactgcagt gaaggtctc
//
