LOCUS       sequence-438730-        5268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  integrative plasmid enabling constitutive expression of mKate2 in 
            Streptococcus pyogenes, integrates into the transcriptionally silent
            locus Spy_1078 via homologous recombination.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5268)
  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 5268)
  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: pEC3115 (pSpy0K6-P23_mKate2~*ssrA)
FEATURES             Location/Qualifiers
     source          1..5268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      829..856
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     867..883
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1923..1939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1923..1939)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2499..3293
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      4415..5003
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4904..4923
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5157..5174
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tgaaaaagac tgagggtcac tttacccgtc aagttgatca ggtcaccaaa tatgacaaag
       61 attatatttg gagtttggtt cattcccaat tacgtgaagg tggtttatca caagctgcta
      121 gtgacttagt catgagctat tttgaagaac gctatgctta tgcgctggaa cacatacggt
      181 tcgcaagaac agctgaagct attgcggaat atgtctttaa tggtgtctta tccgaatgga
      241 ccaagcaact aaggtgacaa gaagcgaagt gagtaagatg ttttggtggc tattattagg
      301 tgtttgggga ctaaggacaa tttggttgat tattgaagtt atcactgcac cagaaatgga
      361 agattatagt tgagtaagga cgttacataa gcagtaaggt ccttttattt tgtcagaaag
      421 gaatgagaga tatgaagttt ttagatttgt ttgcaggaat tggaggcttt cgattgggtt
      481 taattaatca gtgccatgag tgtattggct tttgtgagat tgataagttt gcgaggcaat
      541 cttacaaagc catctatgag accgaaggag aaatagaatt tcatgacatt agacaagtca
      601 cagaccaaga ctttagacaa cttagagggc aagtggatat catctgcggt ggtttttccc
      661 ttgtcaagca ttttcactcg caggcagacg actgggattt gagggcacta ggagaactct
      721 cttctttgag attactcgag cgaccaaaca gataaaaatc atacccataa tcacatccaa
      781 tcaacgccat tgaccgtaat tcagataaaa aattgatatg gaattatggg ctcaccttcg
      841 ggtgggcctt tctgcgtttg gatatagtaa aacgacggcc agtcagcgag accagctgaa
      901 ttcctcgaaa agccctgaca acccttgttc ctaaaaagga ataagcgttc ggtcagtaaa
      961 taatagaaat aaaaaatcag acctaagact gatgacaaaa agagaaaatt ttgataaaat
     1021 agtcttagaa ttaaattaaa aagggaggcc aaatataatg aaaaatatga atgacaatga
     1081 tgttccgcgg tggcggcgat ctaaggagga ataaaaaatg gtttcagaac ttatcaaaga
     1141 aaacatgcac atgaaacttt acatggaagg tactgttaac aaccaccact tcaaatgtac
     1201 ttcagaaggt gaaggtaaac catacgaagg tactcaaact atgcgtatca aagctgttga
     1261 aggtggtcca cttccattcg ctttcgacat ccttgctact tcattcatgt acggttcaaa
     1321 aactttcatc aaccacactc aaggtatccc agacttcttc aaacaatcat tcccagaagg
     1381 tttcacttgg gaacgtgtta ctacttacga agacggtggt gttcttactg ctactcaaga
     1441 cacttcactt caagacggtt gtcttatcta caacgttaaa atccgtggtg ttaacttccc
     1501 atcaaacggt ccagttatgc aaaaaaaaac tcttggttgg gaagcttcaa ctgaaactct
     1561 ttacccagct gacggtggtc ttgaaggtcg tgctgacatg gctcttaaac ttgttggtgg
     1621 tggtcacctt atctgtaacc ttaaaactac ttaccgttca aaaaaaccag ctaaaaacct
     1681 taaaatgcca ggtgtttact acgttgaccg tcgtcttgaa cgtatcaaag aagctgacaa
     1741 agaaacttac gttgaacaac acgaagttgc tgttgctcgt tactgtgacc ttccatcaaa
     1801 acttggtcac cgtgcataaa atacaaactc ttacgcttta gctgcctaag atcgatctgg
     1861 cttcttgcgt aaaaaaggcc cacaaaagtg ggcctttttt ctcggatctg aaggtctcat
     1921 gtgtcatagc tgtttcctgc ttaacaatcc caaaacttgt cgacgataaa cccagcgaac
     1981 catttgaggt gataggtaag attataccga ggtatgaaaa cgagaattgg acctttacag
     2041 aattactcta tgaagcgcca tatttaaaaa gctaccaaga cgaagaggat gaagaggatg
     2101 aggaggcaga ttgccttgaa tatattgaca atactgataa gataatatat cttttatata
     2161 gaagatatcg ccgtatgtaa ggatttcagg gggcaaggca taggcagcgc gcttatcaat
     2221 atatctatag aatgggcaaa gcataaaaac ttgcatggac taatgcttga aacccaggac
     2281 aataacctta tagcttgtaa attctatcat aattgtggtt tcaaaatcgg ctccgtcgat
     2341 actatgttat acgccaactt tgaaaacaac tttgaaaaag ctgttttctg gtatttaagg
     2401 ttttagaatg caaggaacag tgaattggag ttcgtcttgt tataattagc ttcttggggt
     2461 atctttaaat actgtagaaa agaggaagga aataataaat ggctaaaatg agaatatcac
     2521 cggaattgaa aaaactgatc gaaaaatacc gctgcgtaaa agatacggaa ggaatgtctc
     2581 ctgctaaggt atataagctg gtgggagaaa atgaaaacct atatttaaaa atgacggaca
     2641 gccggtataa agggaccacc tatgatgtgg aacgggaaaa ggacatgatg ctatggctgg
     2701 aaggaaagct gcctgttcca aaggtcctgc actttgaacg gcatgatggc tggagcaatc
     2761 tgctcatgag tgaggccgat ggcgtccttt gctcggaaga gtatgaagat gaacaaagcc
     2821 ctgaaaagat tatcgagctg tatgcggagt gcatcaggct ctttcactcc atcgacatat
     2881 cggattgtcc ctatacgaat agcttagaca gccgcttagc cgaattggat tacttactga
     2941 ataacgatct ggccgatgtg gattgcgaaa actgggaaga agacactcca tttaaagatc
     3001 cgcgcgagct gtatgatttt ttaaagacgg aaaagcccga agaggaactt gtcttttccc
     3061 acggcgacct gggagacagc aacatctttg tgaaagatgg caaagtaagt ggctttattg
     3121 atcttgggag aagcggcagg gcggacaagt ggtatgacat tgccttctgc gtccggtcga
     3181 tcagggagga tatcggggaa gaacagtatg tcgagctatt ttttgactta ctggggatca
     3241 agcctgattg ggagaaaata aaatattata ttttactgga tgaattgttt tagtacctag
     3301 atttagatgt ctaaaaagct ttaactacaa gctttttaga catctaatct tttctgaagt
     3361 acatccgcaa ctgtccatac tctgatgttt tatatctttt ctaaaagttc gctagatagg
     3421 gtgaatctaa ccattgactt aaaacaaaaa atgtaacttt tatcctagaa gaagatactc
     3481 aaaagataag tttaggtcat aaaaaaataa gcgataagaa attatatgat gtcaattttt
     3541 tcaagattac tttaaaaata aggaagtcgg tgtttcagat cgattagaaa gtctacagaa
     3601 acagtacatg cttttcaaga agaacgagat aaagaggagg tatccactga agagagtaag
     3661 gaagtcttca agaaatacaa ggggcaacga gatgccatcc atgagtttga agttgaactt
     3721 gtggaacatc aaattgagag gttaattgct gaggacattt atatcaaggt gtcttttagt
     3781 gtaaagcaga gtggtcttat tttcattcct aactatcaat tggatattag gaaagaagaa
     3841 aatcataaaa aatataaagt ctatattcgt gagaccgcac aattcttcat ttacaacaag
     3901 gaggcgtctg aacttaatcg ctatatgcga gggcatgagc tcatttgcca gttaaccaat
     3961 gatagcaaga gtattccaaa aagaagacga caaaccattg atacacttaa gaaaaagatt
     4021 gaagaaataa gtctcctcat tgagttagat acggagaata agccttatca agatattaaa
     4081 gatgacattg ttaaagatat ggctcaatta gatttgacca ttacagagtt gcaggaccat
     4141 attgctcatc tcaataaggt tgcggaggtc ttgcttaatc tgaacaacaa cgatatagag
     4201 aaccgccgtt tggccagata tgactatgcc aagatgaact tgacctgtca gaccaagttt
     4261 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4321 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4381 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4441 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4501 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4561 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4621 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4681 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4741 gttcgtgcac acagcccagc tttgagcgaa cgacctacac cgaactgaga tacctacagc
     4801 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4861 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4921 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4981 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5041 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5101 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5161 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5221 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaag
//
