LOCUS       sequence-438746-        5391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  integrative plasmid enabling constitutive expression of a 
            destabilized mNeongreen reporter in Streptococcus pyogenes, targets 
            the attP site of phage T12 in several S. pyogenes strains via an 
            integrase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5391)
  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 5391)
  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: pEC2935 (p7INT-P23_mNeongreen~ssrA (LDD variant))
FEATURES             Location/Qualifiers
     source          1..5391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(863..881)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(864..881)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(899..915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(899..915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(912..934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    923..939
                     /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)."
     promoter        complement(join(947..953,954..971,972..977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    992..1013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1130..1147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1301..1889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1381..1400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5078..5100
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5092..5109
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5093..5109
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5116..5135
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5116..5134
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 caacatggct tcacttccag ctactcacga acttcacatc ttcggttcaa tcaacggtgt
       61 tgacttcgac atggttggtc aaggtactgg taacccaaac gacggttacg aagaacttaa
      121 ccttaaatca actaaaggtg accttcaatt ctcaccatgg atccttgttc cacacatcgg
      181 ttacggtttc caccaatacc ttccataccc agacggtatg tcaccattcc aagctgctat
      241 ggttgacggt tcaggttacc aagttcaccg tactatgcaa ttcgaagacg gtgcttcact
      301 tactgttaac taccgttaca cttacgaagg ttcacacatc aaaggtgaag ctcaagttaa
      361 aggtactggt ttcccagctg acggtccagt tatgactaac tcacttactg ctgctgactg
      421 gtgtcgttca aaaaaaactt acccaaacga caaaactatc atctcaactt tcaaatggtc
      481 atacactact ggtaacggta aacgttaccg ttcaactgct cgtactactt acactttcgc
      541 taaaccaatg gctgctaact accttaaaaa ccaaccaatg tacgttttcc gtaaaactga
      601 acttaaacac tcaaaaactg aacttaactt caaagaatgg caaaaagctt tcactgacgt
      661 tatgggtatg gacgaacttt acaaagcaaa aaatacaaac tcttacgctt tagatgatta
      721 agatcgatct ggcttcttgc gtaaaaaagg cccacaaaag tgggcctttt ttctcggatc
      781 gatcgatcct gcaggcatgc aagcttggcg taatcatggg atccggagag ctcccaacgc
      841 gttggatgca tagcttgagt attctatagt gtcacctaaa tagcttggcg taatcatggt
      901 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      961 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
     1021 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
     1081 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg
     1141 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1201 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     1321 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1741 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1981 tcttcaccta gatccttttg atcccccaca aaaagaaaaa cgaaatgata caccaatcag
     2041 tgcaaaaaaa gatataatgg gagataagac ggttcgtgtt cgtgctgact tgcaccatat
     2101 cataaaaatc gaaacagcaa agaatggcgg aaacgtaaaa gaagttatgg aaataagact
     2161 tagaagcaaa cttaagagtg tgttgatagt gcagtatctt aaaattttgt ataataggaa
     2221 ttgaagttaa attagatgct aaaaatttgt aattaagaag gagtgattac atgaacaaaa
     2281 atataaaata ttctcaaaac tttttaacga gtgaaaaagt actcaaccaa ataataaaac
     2341 aattgaattt aaaagaaacc gataccgttt acgaaattgg aacaggtaaa gggcatttaa
     2401 cgacgaaact ggctaaaata agtaaacagg taacgtctat tgaattagac agtcatctat
     2461 tcaacttatc gtcagaaaaa ttaaaactga atactcgtgt cactttaatt caccaagata
     2521 ttctacagtt tcaattccct aacaaacaga ggtataaaat tgttgggagt attccttacc
     2581 atttaagcac acaaattatt aaaaaagtgg tttttgaaag tcatgcgtct gacatctatc
     2641 tgattgttga agaaggattc tacaagcgta ccttggatat tcaccgaaca ctagggttgc
     2701 tcttgcacac tcaagtctcg attcagcaat tgcttaagct gccagcggaa tgctttcatc
     2761 ctaaaccaaa agtaaacagt gtcttaataa aacttacccg ccataccaca gatgttccag
     2821 ataaatattg gaagctatat acgtactttg tttcaaaatg ggtcaatcga gaatatcgtc
     2881 aactgtttac taaaaatcag tttcatcaag caatgaaaca cgccaaagta aacaatttaa
     2941 gtaccgttac ttatgagcaa gtattgtcta tttttaatag ttatctatta tttaacggga
     3001 ggaaataatt ctatgagtcg cttttgtaaa tttggaaagt tacacgttac taaagggaat
     3061 gtagataaat tattaggtaa atctaaaaca aaaagcccca cgctctcaaa gtttggcgac
     3121 tctgagcgtg aggcaagaca gtataagaaa caaccattaa aaaggtcatt ttcttgtacc
     3181 tattttatca aattgaaaga tggtatgcaa tgaaaattaa atcatataaa aaggaaaatg
     3241 gtgaaactgc ttataaattt cttttgtatg ccggttatgt taatggaaag agaaaatata
     3301 ttaggcgaga aggtttcaaa actaagcagg ctgcaaggga aaccttaatt agtttacaag
     3361 ctgaacttga taaacctaaa tcaagtatga catttggagc attgacagat caatggctaa
     3421 aggaatatga aaaaaccgtt cagggcagta cctacttaaa aacagaaaga aatattaata
     3481 aacatatttt gccaaaactt gataaagtga agattggaga catcaatcca ctacttatcc
     3541 agcggcttac tgaagaatgg tgcaacgatt taaaatatgg aggaaaaatt cttgggcttg
     3601 ttaggaatat cttaaatcta gctgttagat acggatatat caataacaat ccagctttgc
     3661 caattacacc tccaaaaata aaaaggaaaa gaaaaatgaa taataatttt tatacacttg
     3721 atcaacttaa acaattcctt gaactagttg aaaaaactga caacattgaa aaaatagcct
     3781 tgtttagatt attagcattt actggaatac gaaaagggga gcttctggca ctaacttggg
     3841 atgatttgaa tggtaatact ctatcaatta ataaagctgt cacacgtact caagttggac
     3901 tagaaataga tgttacgaag acaaaatcaa gcgatagatt aatcagctta gatgatgaaa
     3961 ctttggaaat tttacaagaa cttcatgaaa cttttcctac ttctactctt atgttccaat
     4021 ctaaatcagg tggaattatg acgccaagtt taccacgaaa atggctattg caaattatca
     4081 aagggataga cttaccacaa atcacaattc atggtttcag gcacactcat gcaagcttac
     4141 ttttcgaatc aggtctatcc ttgaaacagg tgcaacatag attagggcat ggagatttac
     4201 agacaactat gaacgtatat actcacatca cgcaatcggc aattgatgac attggaacta
     4261 aattcaatca atttgttact aacaagcaac tagattgaca actaattctc aacaaacgtt
     4321 aatttaacaa cattcaagta actcccacca gctccatcaa tgcttaccgt aagtaatcat
     4381 aacttactaa aaccttgtta catcaaggtt ttttcttttt gtcttgttca tgagttacca
     4441 taactttcta tattattgac aactaaattg acaactcttc aattattttt ctgtctactc
     4501 aaagttttct tcatttgata tagtctaatt ccaccatcac ttcttccacc ttctcctctg
     4561 tcacaacttc tccatttctc gcttcctcat gcggcaacac ataatcaaaa atctgtccgt
     4621 ttttacgcac gatcataact gtgtctccta aaatataccc cttatcaatc gcttctttaa
     4681 actcatctat atataacata tttcatcctc ctacctatct attcgtaaaa agataaaaat
     4741 aactattgtt ttgtttgtta ttttataata aaattattaa tataagttaa tgttttttaa
     4801 aaatatacaa ttttattcta tttatagtta gctatttttt cattgttagt aatattggtg
     4861 aattgtaata acctttttta ctactgacag cttccaagga gctaaagagg tccctagggg
     4921 taccgagctc gaattattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt
     4981 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     5041 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     5101 acggccagtg aattgtaata cgactcacta tagggcgaat tgcgaattgg agctctcgaa
     5161 aagccctgac aacccttgtt cctaaaaagg aataagcgtt cggtcagtaa ataatagaaa
     5221 taaaaaatca gacctaagac tgatgacaaa aagagaaaat tttgataaaa tagtcttaga
     5281 attaaattaa aaagggaggc caaatataat gaaaaatatg aatgacaatg atgttccgcg
     5341 gtggcggcga tctaaggagg aataaaaaat ggtttcaaaa ggtgaagaag a
//
