LOCUS       sequence-438731-        5599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  integrative plasmid enabling constitutive expression of lacI in 
            Streptococcus, 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 5599)
  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 5599)
  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: pEC3079 (p7INT-Pveg_lacI)
FEATURES             Location/Qualifiers
     source          1..5599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3177..3199
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3191..3208
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3192..3208
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3215..3234
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3215..3233
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4561..4579)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4562..4579)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4597..4613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4597..4613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4610..4632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4621..4637
                     /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(4645..4651,4652..4669,4670..4675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4690..4711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4828..4845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4999..5587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5079..5098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
       61 atgagattat caaaaaggat cttcacctag atccttttga tcccccacaa aaagaaaaac
      121 gaaatgatac accaatcagt gcaaaaaaag atataatggg agataagacg gttcgtgttc
      181 gtgctgactt gcaccatatc ataaaaatcg aaacagcaaa gaatggcgga aacgtaaaag
      241 aagttatgga aataagactt agaagcaaac ttaagagtgt gttgatagtg cagtatctta
      301 aaattttgta taataggaat tgaagttaaa ttagatgcta aaaatttgta attaagaagg
      361 agtgattaca tgaacaaaaa tataaaatat tctcaaaact ttttaacgag tgaaaaagta
      421 ctcaaccaaa taataaaaca attgaattta aaagaaaccg ataccgttta cgaaattgga
      481 acaggtaaag ggcatttaac gacgaaactg gctaaaataa gtaaacaggt aacgtctatt
      541 gaattagaca gtcatctatt caacttatcg tcagaaaaat taaaactgaa tactcgtgtc
      601 actttaattc accaagatat tctacagttt caattcccta acaaacagag gtataaaatt
      661 gttgggagta ttccttacca tttaagcaca caaattatta aaaaagtggt ttttgaaagc
      721 catgcgtctg acatctatct gattgttgaa gaaggattct acaagcgtac cttggatatt
      781 caccgaacac tagggttgct cttgcacact caagtctcga ttcagcaatt gcttaagctg
      841 ccagcggaat gctttcatcc taaaccaaaa gtaaacagtg tcttaataaa acttacccgc
      901 cataccacag atgttccaga taaatattgg aagctatata cgtactttgt ttcaaaatgg
      961 gtcaatcgag aatatcgtca actgtttact aaaaatcagt ttcatcaagc aatgaaacac
     1021 gccaaagtaa acaatttaag taccgttact tatgagcaag tattgtctat ttttaatagt
     1081 tatctattat ttaacgggag gaaataattc tatgagtcgc ttttgtaaat ttggaaagtt
     1141 acacgttact aaagggaatg tagataaatt attaggtaaa tctaaaacaa aaagccccac
     1201 gctctcaaag tttggcgact ctgagcgtga ggcaagacag tataagaaac aaccattaaa
     1261 aaggtcattt tcttgtacct attttatcaa attgaaagat ggtatgcaat gaaaattaaa
     1321 tcatataaaa aggaaaatgg tgaaactgct tataaatttc ttttgtatgc cggttatgtt
     1381 aatggaaaga gaaaatatat taggcgagaa ggtttcaaaa ctaagcaggc tgcaagggaa
     1441 accttaatta gtttacaagc tgaacttgat aaacctaaat caagtatgac atttggagca
     1501 ttgacagatc aatggctaaa ggaatatgaa aaaaccgttc agggcagtac ctacttaaaa
     1561 acagaaagaa atattaataa acatattttg ccaaaacttg ataaagtgaa gattggagac
     1621 atcaatccac tacttatcca gcggcttact gaagaatggt gcaacgattt aaaatatgga
     1681 ggaaaaattc ttgggcttgt taggaatatc ttaaatctag ctgttagata cggatatatc
     1741 aataacaatc cagctttgcc aattacacct ccaaaaataa aaaggaaaag aaaaatgaat
     1801 aataattttt atacacttga tcaacttaaa caattccttg aactagttga aaaaactgac
     1861 aacattgaaa aaatagcctt gtttagatta ttagcattta ctggaatacg aaaaggggag
     1921 cttctggcac taacttggga tgatttgaat ggtaatactc tatcaattaa taaagctgtc
     1981 acacgtactc aagttggact agaaatagat gttacgaaga caaaatcaag cgatagatta
     2041 atcagcttag atgatgaaac tttggaaatt ttacaagaac ttcatgaaac ttttcctact
     2101 tctactctta tgttccaatc taaatcaggt ggaattatga cgccaagttt accacgaaaa
     2161 tggctattgc aaattatcaa agggatagac ttaccacaaa tcacaattca tggtttcagg
     2221 cacactcatg caagcttact tttcgaatca ggtctatcct tgaaacaggt gcaacataga
     2281 ttagggcatg gagatttaca gacaactatg aacgtatata ctcacatcac gcaatcggca
     2341 attgatgaca ttggaactaa attcaatcaa tttgttacta acaagcaact agattgacaa
     2401 ctaattctca acaaacgtta atttaacaac attcaagtaa ctcccaccag ctccatcaat
     2461 gcttaccgta agtaatcata acttactaaa accttgttac atcaaggttt tttctttttg
     2521 tcttgttcat gagttaccat aactttctat attattgaca actaaattga caactcttca
     2581 attatttttc tgtctactca aagttttctt catttgatat agtctaattc caccatcact
     2641 tcttccacct tctcctctgt cacaacttct ccatttctcg cttcctcatg cggcaacaca
     2701 taatcaaaaa tctgtccgtt tttacgcacg atcataactg tgtctcctaa aatatacccc
     2761 ttatcaatcg cttctttaaa ctcatctata tataacatat ttcatcctcc tacctatcta
     2821 ttcgtaaaaa gataaaaata actattgttt tgtttgttat tttataataa aattattaat
     2881 ataagttaat gttttttaaa aatatacaat tttattctat ttatagttag ctattttttc
     2941 attgttagta atattggtga attgtaataa ccttttttac tactgacagc ttccaaggag
     3001 ctaaagaggt ccctaggggt accgagctcg aattattaac gcttacaatt tccattcgcc
     3061 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     3121 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     3181 gtcacgacgt tgtaaaacga cggccagtga attgtaatac gactcactat aggggcgggt
     3241 tttttatttt acataatgat acataattta ccgaaacttg cggaacataa ttgaggaatc
     3301 atagaatttt gtcaaaataa ttttattgac aacgtcttat taacgttgat ataatttaaa
     3361 ttttatttga caaaaatggg ctcgtgttgt acaataaatg tagttaaagg aggaataaaa
     3421 aatgaaacca gtcacccttt acgatgtcgc agaatacgca ggagtctctt accaaacagt
     3481 cagtcgtgta gtcaatcagg ctagtcatgt atcagcaaaa actcgtgaaa aggttgaagc
     3541 tgcaatggct gaacttaatt acatcccaaa tcgtgttgcc caacagttgg ctggtaaaca
     3601 aagtttgctt atcggagttg ctacctcttc attggcactt catgcccctt ctcaaattgt
     3661 tgccgctatc aaatctcgtg cagatcaatt gggtgcaagt gttgtcgtat ctatggttga
     3721 acgtagtgga gtcgaagcct gtaaggcagc cgtccacaat ttattggctc aacgtgtatc
     3781 tggtttaatt atcaattacc cattggatga ccaagatgcc attgctgtcg aagctgcatg
     3841 taccaatgta ccagcattat ttttggatgt ttcagatcaa acacctatca attcaatcat
     3901 ctttagtcat gaagatggta ctcgtttggg agtcgaacat ttggtagctc ttggtcacca
     3961 acagattgct cttttagcag gaccacttag ttctgtttca gcccgtcttc gtttagctgg
     4021 ttggcacaaa tacttaactc gtaatcaaat tcagcctatc gctgaacgtg agggtgattg
     4081 gtctgcaatg tcaggatttc aacagacaat gcaaatgttg aatgaaggta ttgtaccaac
     4141 tgcaatgttg gttgccaatg accaaatggc ccttggagct atgcgtgcaa ttaccgaatc
     4201 aggtttgcgt gttggagctg atattagtgt tgtcggttac gatgatacag aagattcaag
     4261 ttgttacatt ccacctttga caactatcaa gcaagatttt cgtttgcttg gacagacaag
     4321 tgttgatcgt ttattgcaac tttctcaggg tcaggctgtc aagggaaatc agcttttacc
     4381 tgtatctctt gttaagcgta agaccacatt agcaccaaat actcaaaccg cctcacctcg
     4441 tgccttggca gatagtctta tgcagcttgc acgtcaggtc tctcgtttgg aaagtggaca
     4501 ataagatcga tcgatcaaaa aaggcccaca aaagtgggcc ttttttcata gcttgagtat
     4561 tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     4621 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     4681 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     4741 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     4801 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     4861 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4921 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4981 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5041 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5101 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     5161 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5221 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5281 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5341 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5401 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5461 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5521 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5581 atctcaagaa gatcctttg
//
