LOCUS       sequence-125541-        7813 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modular vector for secreted luciferase reporter expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7813)
  AUTHORS   Oliveira Paiva AM, Friggen AH, Hossein-Javaheri S, Smits WK
  TITLE     The Signal Sequence of the Abundant Extracellular Metalloprotease 
            PPEP-1 Can Be Used to Secrete Synthetic Reporter Proteins in 
            Clostridium difficile.
  JOURNAL   ACS Synth Biol. 2016 Jun 23.
  PUBMED    27333161
REFERENCE   2  (bases 1 to 7813)
  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: "ACS Synth 
            Biol. 2016 Jun 23."
COMMENT     Sequence Label: pAP24
FEATURES             Location/Qualifiers
     source          1..7813
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(982..1353)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            1386..1495
                     /note="incP origin of transfer"
     primer_bind     complement(2684..2701)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2855..3443)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2935..2954)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7019..7645)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     7550..7569
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    7752..7770
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
ORIGIN
        1 cctgcagtaa aggagaaaat tttatgagac caagtaaaaa attattaata gctataatat
       61 caatattctt aataagttca gttccagtaa gtgcacatgc agatagtgtt tttacacttg
      121 aagattttgt gggtgattgg agacaaactg ctggatataa tttagatcaa gttttagaac
      181 agggtggagt gagttctctt tttcaaaatc ttggtgtctc agttacacca atacaaagaa
      241 tagttttaag tggagaaaat ggattaaaga tcgatataca tgtaataatt ccttatgagg
      301 gactatcagg ggaccaaatg ggacaaattg agaaaatatt caaagttgta tatccagtag
      361 acgatcatca ctttaaagta atattacatt atggaacttt agtaattgat ggtgtaacac
      421 caaatatgat agattatttt ggtagaccat acgaagggat tgcagttttt gatggaaaga
      481 aaataaccgt aacaggcact ttgtggaatg gaaataaaat aatagatgaa aggttaatta
      541 atcctgatgg atctttattg tttagagtaa ctataaacgg tgttacaggt tggagacttt
      601 gtgaacgtat tctagcatag ggatcctata agttttaata aaactttaaa tagaaaaagg
      661 cttctctcat gagaagtctt ttttatttaa aataaatata aaataaaata gaggctataa
      721 atagcctcta ttttatgtga gaaatcccta aataaaaaga tgccagtgtg ctggaattcc
      781 ccttcaatat tccacaataa ggctgtaggc gctagggacc tctttagctc cttggaagct
      841 gtcagtagta ggggatcggt cttgccttgc tcgtcggtga tgtacttcac cagctccgcg
      901 aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg caatcacgcg caccccccgg
      961 ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg cggaccacgc ccatcatgac
     1021 cttgccaagc tcgtcctgct tctcttcgat cttcgccagc agggcgagga tcgtggcatc
     1081 accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc cgcccaggcg
     1141 gcccaggtcg ccattgatgc gggccagctc gcggacgtgc tcatagtcca cgacgcccgt
     1201 gattttgtag ccctggccga cggccagcag gtaggccgac aggctcatgc cggccgccgc
     1261 cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga taggtgggct
     1321 gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg ttacgccggc
     1381 ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt gcgaataagg
     1441 gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc tgcccggctg
     1501 acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc ctgtatatcg
     1561 tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc aggaagggcg
     1621 aattctgcag atctgaccgg tctctgaaaa tataaaaacc acagattgat actaaaacct
     1681 tggttgtgtt gcttttcggg gcttaaatca aggaaaaatc cttgttttaa gcctttcaaa
     1741 aagaaacaca aggtctttgt actaacctgt ggttatgtat aaaattgtag attttagggt
     1801 aacaaaaaac accgtatttc tacgatgttt ttgcttaaat acttgttttt agttacagac
     1861 aaacctgaag ttaactattt atcaattcct gcaattcgtt tacaaaacgg caaatgtgaa
     1921 atccgtcaca tactgcgtga tgaacttgaa ttgccaaagg aagtataatt ttgttatctt
     1981 ctttataata tttccccata gtaaaaatag gaatcaaata atcatatcct ttctgcaaat
     2041 tcagattaaa gccatcgaag gttgaccacg gtatcataga tacattaaaa atgttttccg
     2101 gagcatttgg ctttccttcc attctatgat tgtttccata ccgttgcgta tcactttcat
     2161 aatctgctaa aaatgattta aagtcagact tacactcagt ccaaaggctg gaaaatgttt
     2221 cagtatcatt gtgaaatatt gtatagcttg gtatcatctc atcatatatc cccaattcac
     2281 catcttgatt gattgccgtc ctaaactctg aatggcggtt tacaatcatt gcaatataat
     2341 aaagcattgc aggatatagt ttcattccct tttcctttat ttgtgtgata tccactttaa
     2401 cggtcatgct gtaggtacaa ggtacacttg caaagtagtg gtcaaaatac tcttttctgt
     2461 tccaactatt tttatcaatt ttttcaaata ccatctaagt tccctctcaa attcaagttt
     2521 atcgctctaa tgaacaaaga tattatacca catttttgtg aatttttcaa cttgcccact
     2581 tcgactgcac tcccgactta ataacttctt gaacacttgc cgaaaaagaa aaactgccgg
     2641 gtacgtaccc gggatcgatc cccgccgagc gctcttccgc ttcctcgctc actgactcgc
     2701 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2761 tatccactga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2821 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2881 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2941 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3001 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     3061 gtaggtatca cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3121 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3181 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3241 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     3301 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3361 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3421 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     3481 agtggaacga aaactcacgt taagggattt tggtcatgaa atgcaagttt ctaactaaca
     3541 tacatcatag ttactaaact atggtaacta taattttatt aactatatta agcactgatt
     3601 agtactataa ctcaatataa gcatatcccc tgtatcgtaa ctagagaacc aaacgacgga
     3661 aaaagcgata tagataagtt tagatagtgc accaattttt tacttttcta ttttagaatc
     3721 tagcatttcc aatgcttact aatactattt ttgattttta acttgtttta aaaaagcacc
     3781 tactaattag gattctagct ctatattcta gttatacatt ctctatattc tagctctata
     3841 ttctagttat acattctcta tattctagct ctatattcta gttatacatt ctctatattc
     3901 tagctctata ttctagttat acattctcta tattctagct ctatattcta gttatacatt
     3961 ctctatattc tagctctata ttctagttat acattctcta tattctagct ctatattcta
     4021 gttatacatt ctctaaaaaa catatatata tatactacat atatatgtac aattatttta
     4081 ccaaaagaaa tttttaaaac ggcttaattt caatactttg atgccatttt ttttttgcaa
     4141 ttctcgtaca attattttac caaaagaaat ttttattaat tagttttttt gcataaaaat
     4201 aggactagaa aatcaattct aatcctatca aaaatctact tgtttaatct atatgtatag
     4261 tacataaatc ctctatcatt tattttagtt tcccatacat ctaaaatttt agtatctaat
     4321 aaagattcta tcatatttga aattctttgc tgtgttactc ctaaatcttt tgctaattcc
     4381 tcttgattaa ttcttaatat atttcctttt attttatcat ctttgaccaa taaatcttgc
     4441 ttatatctca tatgagtata taatcttaat tcttcagttg aaattattcc ccaaattact
     4501 gccatggtaa caaagtaact aatagtatat tgtttatcta tattacatct aacttctttt
     4561 acattgtatg tatcttttat tcctagcttt gtaacacctt ttgtttttgt aatataatca
     4621 ttatgttgaa gttcttttaa tgtttctctt aatgtctttt cactcataac aatgttttta
     4681 actttcttct tacgtttata ggttatcagc tccattatat cgtctatgtt taattctcta
     4741 tctttattgt taagtaacac attatagata aacaattgat taccagtcat tattttaacc
     4801 cctttcctat tcttatattt caaatccttt gagtgcttat gtggcatatt gaacaaagta
     4861 tcttcatcac tatatataaa atctgattct attttattcc aacaatcact attaaactca
     4921 caacctgtac aaccaaaatt aaccgtttta agcttctcat acatatactt gacttgatat
     4981 tctagctcac tatttgaaag actattttgt gatatatagt tccattctct agcaacggat
     5041 attacttgac ttaaactttt attcctcaat cttaaagtca caactatttt ttgcaaatct
     5101 aggtttttat gtccatcttt actaccattt tttaaaatat ctactataca atttggtagc
     5161 ttactatcta atatatattt tgtattagta ttacctactg tttttacatc tttgcaatat
     5221 tttttagcta acttttctat gtcatatcta tatatatttt tgtcttggtg tattattttt
     5281 acttgtttag tagtattttt aatattatat gtataaggta ctcttaatac ttgagtagat
     5341 aaatttgctt gtttatctgc acccaattta tttattaatg tatttgtaac ttctgatact
     5401 aacttaatgt tattagtttt attaattagc acataaacat ggaatccatt tccactatca
     5461 actaggatat gataatgtaa tcctatctta gtaaataaat taattatatc tttataatta
     5521 aaaccttgtc ctaattcttt tttgtcaaaa tcaaatccta atgcatatct atttattaag
     5581 ttttcattgg ttcctgcttt tccatcagtt gtacataggt taaaaaatgt attttcataa
     5641 taaatatttt ttgcgaaggt aaattctact atatcatcta tatttttaaa aaaagttgtt
     5701 ttactatatt ctttaaattc ttctccgagt tctaattgtt tactattact tttattattt
     5761 tgaaatactc taacatattc attatcaata agattatttc cttgtcctat accatctcta
     5821 aatactgcca ttagaaattt tttcaactta tattttgaat ctaattgttc catatattta
     5881 ttatctcctt atctagtttt tttaagagat aacactacat taaatatata cttattgact
     5941 aatcaacttt tacaactatt gaataatttt agcttagtat gtataataat attgttaaga
     6001 tgctaaaatt gatagcaata ttattatata actacagata cttaatcagt tagaccgata
     6061 ttggagttgg tagctccata aactcaattg gtagttgagt tgtcattgtc tagtagtatt
     6121 atctcatttt gttgttatct taaaatatta ttttaaaaat actctgcttg gtcggcttag
     6181 ggtatttttt ttgtttaatt ccaatattgg ttcaaactcc tgtgtaatac ttttgacact
     6241 tccatctgta ttgaattgac agttgatatg gtatctcatt attggcttgt tcttatgttt
     6301 ttctatgtat ttatttttta ctttcaaata atttgtattt ccatttttag ttatgtttaa
     6361 cacactatcc ttattactca ataaaattat tttttgtttt aacatatgat ttatagaatc
     6421 taaaaattga tattgactta taatagtact ttctacctct ctagcttttt ctagtgtttc
     6481 aaattcttca aactctgcta tattgctata gtatagctta tcattaataa tattatctga
     6541 caactttaat tccattttat aaaacaattc caagttatgt ataaaatttt taatgttaca
     6601 acatatagac ttattatttg ttcttatatg gtatctccat ttactacttc tagctctcat
     6661 gttatatgca caactactcc caacataaac atatattgct tgttctaact catatttttg
     6721 taaaatttca acatcatatc cagtcaaatc gtactctctc ttatatacaa atggtttttc
     6781 taattttttg ttcaataaat ctcctcctat ttcttactta atttagctaa aaatctattt
     6841 atatttcacc tcctttttga ctttaagcct acgaatacca tatttatgat tataacatct
     6901 tctatttttc ccaaatcctt acatctcccc taatttttag acttaagggc gaattctgca
     6961 tcaagctagc ataaaaataa gaagcctgca tttgcaggct tcttattttt atggtacctt
     7021 aagacccact ttcacattta agttgttttt ctaatccgca tatgatcaat tcaaggccga
     7081 ataagaaggc tggctctgca ccttggtgat caaataattc gatagcttgt cgtaataatg
     7141 gcggcatact atcagtagta ggtgtttccc tttcttcttt agcgacttga tgctcttgat
     7201 cttccaatac gcaacctaaa gtaaaatgcc ccacagcgct gagtgcatat aatgcattct
     7261 ctagtgaaaa accttgttgg cataaaaagg ctaattgatt ttcgagagtt tcatactgtt
     7321 tttctgtagg ccgtgtacct aaatgtactt ttgctccatc gcgatgactt agtaaagcac
     7381 atctaaaact tttagcgtta ttacgtaaaa aatcttgcca gctttcccct tctaaagggc
     7441 aaaagtgagt atggtgccta tctaacatct caatggctaa ggcgtcgagc aaagcccgct
     7501 tattttttac atgccaatac aatgtaggct gctctacacc tagcttctgg gcgagtttac
     7561 gggttgttaa accttcgatt ccgacctcat taagcagctc taatgcgctg ttaatcactt
     7621 tacttttatc taatctagac atcattaatt cctccttttt gttgacatta tatcattgat
     7681 agagttattt gtcaaactag ttttttattt cgatgccctg gacttcatga aaaactaaaa
     7741 aaaatattga cactctatca ttgatagagt ataattaaaa taagcttgat cgtagcgtta
     7801 acagatctga gct
//
