LOCUS       sequence-174987-        8442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Toxin-antitoxin stabilized bioluminescent reporter plasmid for 
            Streptococcus expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8442)
  AUTHORS   Loh JM, Proft T
  TITLE     Toxin-antitoxin-stabilized reporter plasmids for biophotonic imaging
            of Group A streptococcus.
  JOURNAL   Appl Microbiol Biotechnol. 2013 Nov;97(22):9737-45. doi: 
            10.1007/s00253-013-5200-7. Epub 2013 Sep 6.
  PUBMED    24061415
REFERENCE   2  (bases 1 to 8442)
  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"; doi: 
            "10.1007/s00253-013-5200-7"; journalName: "Appl Microbiol 
            Biotechnol"; date: "2013-11"; volume: "97"; issue: "22"; pages: 
            "9737-45"
COMMENT     Sequence Label: pLZ12Km2-P23R:TA:ffluc
FEATURES             Location/Qualifiers
     source          1..8442
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     472..494
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     486..503
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     487..503
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     513..532
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        513..531
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             759..2411
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(825..843)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(4617..4633)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4618..4634)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(5456..5467)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             complement(7108..7902)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 aagttttggg attgttaagg gttccgaggc tcaacgtcaa taaagcaatt ggaataaaga
       61 agcgaaaaag gagaagtcgg ttcagaaaaa gaaggatatg gatctggagc tgtaatataa
      121 aaaccttctt caactaacgg ggcaggttag tgacattaga aaaccgactg taaaaagtac
      181 agtcggcatt atctcatatt ataaaagcca gtcattaggc ctatctgaca attcctgaat
      241 agagttcata aacaatcctg catgataacc atcacaaaca gaatgatgta cctgtaaaga
      301 tagcggtaaa tatattgaat tacctttatt aatgaatttt cctgctgtaa taatgggtag
      361 aaggtaatta ctattattat tgatatttaa gttaaaccca gtaaatgaag tccatgctgg
      421 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
      481 gacgttgtaa aacgacggcc agtgagcgcg cgtaatacga ctcactatag ggcgaattgg
      541 agctctcgaa aagccctgac aacccttgtt cctaaaaagg aataagcgtt cggtcagtaa
      601 ataatagaaa taaaaaatca gacctaagac tgatgacaaa aagagaaaat tttgataaaa
      661 tagtcttaga attaaattaa aaagggaggc caaatataat gaaaaatatg aatgacaatg
      721 atgttccgcg gtggcggcga tctaaggagg aataaaaaat ggaagacgcc aaaaacataa
      781 agaaaggccc ggcgccattc tatcctctag aggatggaac cgctggagag caactgcata
      841 aggctatgaa gagatacgcc ctggttcctg gaacaattgc ttttacagat gcacatatcg
      901 aggtgaacat cacgtacgcg gaatacttcg aaatgtccgt tcggttggca gaagctatga
      961 aacgatatgg gctgaataca aatcacagaa tcgtcgtatg cagtgaaaac tctcttcaat
     1021 tctttatgcc ggtgttgggc gcgttattta tcggagttgc agttgcgccc gcgaacgaca
     1081 tttataatga acgtgaattg ctcaacagta tgaacatttc gcagcctacc gtagtgtttg
     1141 tttccaaaaa ggggttgcaa aaaattttga acgtgcaaaa aaaattacca ataatccaga
     1201 aaattattat catggattct aaaacggatt accagggatt tcagtcgatg tacacgttcg
     1261 tcacatctca tctacctccc ggttttaatg aatacgattt tgtaccagag tcctttgatc
     1321 gtgacaaaac aattgcactg ataatgaatt cctctggatc tactgggtta cctaagggtg
     1381 tggcccttcc gcatagaact gcctgcgtca gattctcgca tgccagagat cctatttttg
     1441 gcaatcaaat cattccggat actgcgattt taagtgttgt tccattccat cacggttttg
     1501 gaatgtttac tacactcgga tatttgatat gtggatttcg agtcgtctta atgtatagat
     1561 ttgaagaaga gctgttttta cgatcccttc aggattacaa aattcaaagt gcgttgctag
     1621 taccaaccct attttcattc ttcgccaaaa gcactctgat tgacaaatac gatttatcta
     1681 atttacacga aattgcttct gggggcgcac ctctttcgaa agaagtcggg gaagcggttg
     1741 caaaacgctt ccatcttcca gggatacgac aaggatatgg gctcactgag actacatcag
     1801 ctattctgat tacacccgag ggggatgata aaccgggcgc ggtcggtaaa gttgttccat
     1861 tttttgaagc gaaggttgtg gatctggata ccgggaaaac gctgggcgtt aatcagagag
     1921 gcgaattatg tgtcagagga cctatgatta tgtccggtta tgtaaacaat ccggaagcga
     1981 ccaacgcctt gattgacaag gatggatggc tacattctgg agacatagct tactgggacg
     2041 aagacgaaca cttcttcata gttgaccgct tgaagtcttt aattaaatac aaaggatatc
     2101 aggtggcccc cgctgaattg gaatcgatat tgttacaaca ccccaacatc ttcgacgcgg
     2161 gcgtggcagg tcttcccgac gatgacgccg gtgaacttcc cgccgccgtt gttgttttgg
     2221 agcacggaaa gacgatgacg gaaaaagaga tcgtggatta cgtcgccagt caagtaacaa
     2281 ccgcgaaaaa gttgcgcgga ggagttgtgt ttgtggacga agtaccgaaa ggtcttaccg
     2341 gaaaactcga cgcaagaaaa atcagagaga tcctcataaa ggccaagaag ggcggaaagt
     2401 ccaaattgta aggatcggcc gctctagagt cgtgttaatt tttatgaagg cttgaaaaat
     2461 ggaaacttgg cttcttctat tgttcatttg actgataatt tagacttgat ccctggcacg
     2521 tttgatttga tgttactgcc aaaattaact cgttcatgga cgtttgaaaa tgaaagtaga
     2581 ttgcttgcta ctcttttagc acctttaaaa agtgactatg atctcattat tattgatact
     2641 gtaccaacgc caagcgttta tacaaataat gcaatcgtgg cgagtgatta cgttatgatc
     2701 cctttacaag cagaagaaga aagtacaaac aacattcaaa actatatttc ctatttgatt
     2761 gatttacaag aacaatttaa ccctggacta gatatgatcg gttttgttcc ttatttagtt
     2821 gatacggaca gcgcaacgat aaaatcaaac ctggaagaac tgtataagca acataaagaa
     2881 gataacttgg ttttccaaaa tattatcaag cgaagtaata aagtaagtac ctggtctaaa
     2941 aacggcatta cagagcataa aggctatgac aaaaaagttt tatccatgta taagaacgta
     3001 ttttttgaaa tgcttgagcg aattattcaa ttagaaaacg aaaaagaata gaatcacaaa
     3061 tcacaagtga ttaatcacaa atcacttgtg atttgtgatt gttgatgata aaataagagt
     3121 aagaagaaat agaaagaagt gagtgattgt gggaaattta ggcgcacaaa aagcaaaacg
     3181 aaatgataca ccaatcagtg caaaaaaaga tataatggga gataagacgg ttcgtgttcg
     3241 tgctgacttg caccatatca taaaaatcga aacagcaaag aatggcggaa acgtaaaaga
     3301 agttatggat caagccttag aagaatatat acggaaatat ttacctgaca aactttaaaa
     3361 ggagtgaaaa tgaaatggca gttacgtatg aaaaaacatt tgaaatagag atcattaacg
     3421 aattatcggc aagcgtttat aatcgagtat taaactatgt tttgaaccat gaattaaata
     3481 aaaatgactc tcaattattg gaagtcaatt tattaaacca attaaagctt gcaaaacgtg
     3541 taaatctttt tgattattct ttagaagaat tacaagccgt tcatgagtat tggcggtcaa
     3601 tgaatcgtta ctcaaaacaa gttttgaata aagagaaagt ggcttaatat ggcaaatata
     3661 gtcaatttta ctgacaaaca atttgagaat cgcttaaatg ataatttaga agaattgatt
     3721 caaggaaaaa aagcggttga atcgccaacc gcttttttac ttggtgggca accagggtca
     3781 gggaaaacca gtttgcgatc agcaattttt gaagaaacac aagggaatgt tattgtcatt
     3841 gataatgata cctttaaaca acagcaccct aattttgatg aactagtgaa actttatgaa
     3901 aaagacgtag taaaacacgt taccccttat tctaatcgca tgacagaagc gatcataagc
     3961 cgtttgagcg atcaagggta taatttggtg atcgaaggta caggacgaac aacagacgtt
     4021 cctattcaaa ccgcaacaat gcttcaagcc aaaggttatg aaacaaaaat gtatgtcatg
     4081 gcagtaccta aaatcaactc atatttagga acaattgaac gatatgaaac catgtatgca
     4141 gatgatccaa tgacagccag ggcaacacca aaacaagcgc atgatattgt tgtcaaaaac
     4201 ttaccgacca atttagaaac ccttcataaa acgggcttat ttagcgatat aaggctatat
     4261 aacagagaag gagtaaaact ctattcaagt ttggaaacac cttctattag tccaaaagaa
     4321 accttagaaa aagaattgaa tcgtaaagta tcagggaaag aaattcaacc gactttagaa
     4381 cgaatagagc aaaaaatggt tctaaataaa caccaagaga cacctgaatt taaagcaatt
     4441 caacaaaaat tggaaagctt gcagccacct acaccaccaa tacccaaaac acctaaactt
     4501 ccaggtattt aaaccttaaa aaaggaaaag agtagttacc aaaaaacggt aactactctt
     4561 tttttataaa aacatttcct catgttggac ctgcaggaat tcgatatcaa gcttatcgat
     4621 accgtcgacc tcgagggggg gcccatcagt ctgacgacca agagagccat aaacaccaat
     4681 agccttaaca tcatccccat atttatccaa tattcgttcc ttaatttcat gaacaatctt
     4741 cattctttct tctctagtca ttattattgg tccattcact attctcattc ccttttcaga
     4801 taattttaga tttgcttttc taaataagaa tatttggaga gcaccgttct tattcagcta
     4861 ttaataactc gtcttcctaa gcatccttca atccttttaa taacaattat agcatctaat
     4921 cttcaacaaa ctggcccgtt tgttgaacta ctctttaata aaataatttt tccgttccca
     4981 attccacatt gcaataatag aaaatccatc ttcatcggct ttttcgtcat catctgtatg
     5041 aatcaaatcg ccttcttctg tgtcatcaag gtttaatttt ttatgtattt cttttaacaa
     5101 accaccatag gagattaacc ttttacggtg taaaccttcc tccaaatcag acaaacgttt
     5161 caaattcttt tcttcatcat cggtcataaa atccgtatcc tttacaggat attttgcagt
     5221 ttcgtcaatt gccgattgta tatccgattt atatttattt ttcggtcgat tttttattaa
     5281 aacgtctcaa aatcgtttct gagacgtttt agcgtttatt tcgtttagtt atcggcataa
     5341 tcgttaaaac aggcgttatc gtagcgtaaa agcccttgag cgtagcgtgg ctttgcagcg
     5401 aagatgttgt ctgttagatt atgaaagccg atgactgaat gaaataataa gcgcagcgtc
     5461 cttctatttc ggttggagga ggctcaaggg agtttgaggg aatgaaattc cctcatgggt
     5521 ttgattttaa aaattgcttg caattttgcc gagcggtagc gctggaaaat ttttgaaaaa
     5581 aatttggaat ttggaaaaaa atggggggaa aggaagcgaa ttttgcttcc gtactacgac
     5641 cccccattaa gtgccgagtg ccaatttttg tgccaaaaac gctctatccc aactggctca
     5701 agggtttgag gggtttttca atcgccaacg aatcgccaac gttttcgcca acgtttttta
     5761 taaatctata tttaagtagc tttattgttg tttttatgat tacaaagtga tacactaatt
     5821 ttataaaatt atttgattgg agttttttaa atggtgattt cagaatcgaa aaaaagagtt
     5881 atgatttctc tgacaaaaga gcaagataaa aaattaacag atatggcgaa acaaaaaggt
     5941 ttttcaaaat ctgcggttgc ggcgttagct atagaagaat atgcaagaaa ggaatcagaa
     6001 caaaaaaaat aagcgaaagc tcgcgttttt agaaggatac gagttttcgc tacttgtttt
     6061 tgataaggta attatatcat ggctattaaa aatactaaag ctagaaattt tggattttta
     6121 ttatatcctg actcaattcc taatgattgg aaagaaaaat tagagagttt gggcgtatct
     6181 atggctgtca gtcctttaca cgatatggac gaaaaaaaag ataaagatac atggaatagt
     6241 agtgatgtta tacgaaatgg aaagcactat aaaaaaccac actatcacgt tatatatatt
     6301 gcacgaaatc ctgtaacaat agaaagcgtt aggaacaaga ttaagcgaaa attggggaat
     6361 agttcagttg ctcatgttga gatacttgat tatatcaaag gttcatatga atatttgact
     6421 catgaatcaa aggacgctat tgctaagaat aaacatatat acgacaaaaa agatattttg
     6481 aacattaatg attttgatat tgaccgctat ataacacttg atgaaagcca aaaaagaaaa
     6541 ttgaagaatt tacttttaga tatagtggat gactataatt tggtaaatac aaaagattta
     6601 atggctttta ttcgccttag gggagcggag tttggaattt taaatacgaa tgatgtaaaa
     6661 gatattgttt caacaaactc tagcgccttt agattatggt ttgagggcaa ttatcagtgt
     6721 ggatatagag caagttatgc aaaggttctt gatgctgaaa cgggggaaat aaaatgacaa
     6781 acaaagaaaa agagttattt gctgaaaatg aggaattaaa aaaagaaatt aaggacttaa
     6841 aagagcgtat tgaaagatac agagaaatgg aagttgaatt aagtacaaca atagatttat
     6901 tgagaggagg gattattgaa taaataaaag cccccctgac gaaagtcgac gatacaaatt
     6961 cctcgtaggc gctcgggacc cctatctagc gaacttttag aaaagatata aaacatcaga
     7021 gtatggacag ttgcggatgt acttcagaaa agattagatg tctaaaaagc ttgtagttaa
     7081 agctttttag acatctaaat ctaggtacta aaacaattca tccagtaaaa tataatattt
     7141 tattttctcc caatcaggct tgatccccag taagtcaaaa aatagctcga catactgttc
     7201 ttccccgata tcctccctga tcgaccggac gcagaaggca atgtcatacc acttgtccgc
     7261 cctgccgctt ctcccaagat caataaagcc acttactttg ccatctttca caaagatgtt
     7321 gctgtctccc aggtcgccgt gggaaaagac aagttcctct tcgggctttt ccgtctttaa
     7381 aaaatcatac agctcgcgcg gatctttaaa tggagtgtct tcttcccagt tttcgcaatc
     7441 cacatcggcc agatcgttat tcagtaagta atccaattcg gctaagcggc tgtctaagct
     7501 attcgtatag ggacaatccg atatgtcgat ggagtgaaag agcctgatgc actccgcata
     7561 cagctcgata atcttttcag ggctttgttc atcttcatac tcttccgagc aaaggacgcc
     7621 atcggcctca ctcatgagca gattgctcca gccatcatgc cgttcaaagt gcaggacctt
     7681 tggaacaggc agctttcctt ccagccatag catcatgtcc ttttcccgtt ccacatcata
     7741 ggtggtccct ttataccggc tgtccgtcat ttttaaatat aggttttcat tttctcccac
     7801 cagcttatat accttagcag gagacattcc ttccgtatct tttacgcagc ggtatttttc
     7861 gatcagtttt ttcaattccg gtgatattct cattttagcc atttattatt tccttcctct
     7921 tttctacagt atttaaagat accccaagaa gctaattata acaagacgaa ctccaattca
     7981 ctgttccttg cattctaaaa ccttaaatac cagaaaacag ctttttcaaa gttgttttca
     8041 aagttggcgt ataacatagt atcgacggag ccgattttga aaccacaatt atgatagaat
     8101 ttacaagcta taaggttatt gtcctgggtt tcaagcatta gtccatgcaa gtttttatgc
     8161 tttgcccatt ctatagatat attgataagc gcgctgccta tgccttgccc cctgaaatcc
     8221 ttacatacgg cgatatcttc tatataaaag atatattatc ttatcagtat tgtcaatata
     8281 ttcaaggcaa tctgcctcct catcctcttc atcctcttcg tcttggtagc tttttaaata
     8341 tggcgcttca tagagtaatt ctgtaaaggt ccaattctcg ttttcatacc tcggtataat
     8401 cttacctatc acctcaaatg gttcgctggg tttatcgtcg ac
//
