LOCUS       sequence-147069-        9160 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Modular vector for secreted amylase reporter expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9160)
  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 9160)
  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: pWKS1583
FEATURES             Location/Qualifiers
     source          1..9160
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3333..3959)
                     /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     3864..3883
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    4066..4084
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     CDS             complement(6456..6827)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            6860..6969
                     /note="incP origin of transfer"
     primer_bind     complement(8158..8175)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(8329..8917)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(8409..8428)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtgcaccaat tttttacttt tctattttag aatctagcat ttccaatgct tactaatact
       61 atttttgatt tttaacttgt tttaaaaaag cacctactaa ttaggattct agctctatat
      121 tctagttata cattctctat attctagctc tatattctag ttatacattc tctatattct
      181 agctctatat tctagttata cattctctat attctagctc tatattctag ttatacattc
      241 tctatattct agctctatat tctagttata cattctctat attctagctc tatattctag
      301 ttatacattc tctatattct agctctatat tctagttata cattctctaa aaaacatata
      361 tatatatact acatatatat gtacaattat tttaccaaaa gaaattttta aaacggctta
      421 atttcaatac tttgatgcca tttttttttt gcaattctcg tacaattatt ttaccaaaag
      481 aaatttttat taattagttt ttttgcataa aaataggact agaaaatcaa ttctaatcct
      541 atcaaaaatc tacttgttta atctatatgt atagtacata aatcctctat catttatttt
      601 agtttcccat acatctaaaa ttttagtatc taataaagat tctatcatat ttgaaattct
      661 ttgctgtgtt actcctaaat cttttgctaa ttcctcttga ttaattctta atatatttcc
      721 ttttatttta tcatctttga ccaataaatc ttgcttatat ctcatatgag tatataatct
      781 taattcttca gttgaaatta ttccccaaat tactgccatg gtaacaaagt aactaatagt
      841 atattgttta tctatattac atctaacttc ttttacattg tatgtatctt ttattcctag
      901 ctttgtaaca ccttttgttt ttgtaatata atcattatgt tgaagttctt ttaatgtttc
      961 tcttaatgtc ttttcactca taacaatgtt tttaactttc ttcttacgtt tataggttat
     1021 cagctccatt atatcgtcta tgtttaattc tctatcttta ttgttaagta acacattata
     1081 gataaacaat tgattaccag tcattatttt aacccctttc ctattcttat atttcaaatc
     1141 ctttgagtgc ttatgtggca tattgaacaa agtatcttca tcactatata taaaatctga
     1201 ttctatttta ttccaacaat cactattaaa ctcacaacct gtacaaccaa aattaaccgt
     1261 tttaagcttc tcatacatat acttgacttg atattctagc tcactatttg aaagactatt
     1321 ttgtgatata tagttccatt ctctagcaac ggatattact tgacttaaac ttttattcct
     1381 caatcttaaa gtcacaacta ttttttgcaa atctaggttt ttatgtccat ctttactacc
     1441 attttttaaa atatctacta tacaatttgg tagcttacta tctaatatat attttgtatt
     1501 agtattacct actgttttta catctttgca atatttttta gctaactttt ctatgtcata
     1561 tctatatata tttttgtctt ggtgtattat ttttacttgt ttagtagtat ttttaatatt
     1621 atatgtataa ggtactctta atacttgagt agataaattt gcttgtttat ctgcacccaa
     1681 tttatttatt aatgtatttg taacttctga tactaactta atgttattag ttttattaat
     1741 tagcacataa acatggaatc catttccact atcaactagg atatgataat gtaatcctat
     1801 cttagtaaat aaattaatta tatctttata attaaaacct tgtcctaatt cttttttgtc
     1861 aaaatcaaat cctaatgcat atctatttat taagttttca ttggttcctg cttttccatc
     1921 agttgtacat aggttaaaaa atgtattttc ataataaata ttttttgcga aggtaaattc
     1981 tactatatca tctatatttt taaaaaaagt tgttttacta tattctttaa attcttctcc
     2041 gagttctaat tgtttactat tacttttatt attttgaaat actctaacat attcattatc
     2101 aataagatta tttccttgtc ctataccatc tctaaatact gccattagaa attttttcaa
     2161 cttatatttt gaatctaatt gttccatata tttattatct ccttatctag tttttttaag
     2221 agataacact acattaaata tatacttatt gactaatcaa cttttacaac tattgaataa
     2281 ttttagctta gtatgtataa taatattgtt aagatgctaa aattgatagc aatattatta
     2341 tataactaca gatacttaat cagttagacc gatattggag ttggtagctc cataaactca
     2401 attggtagtt gagttgtcat tgtctagtag tattatctca ttttgttgtt atcttaaaat
     2461 attattttaa aaatactctg cttggtcggc ttagggtatt ttttttgttt aattccaata
     2521 ttggttcaaa ctcctgtgta atacttttga cacttccatc tgtattgaat tgacagttga
     2581 tatggtatct cattattggc ttgttcttat gtttttctat gtatttattt tttactttca
     2641 aataatttgt atttccattt ttagttatgt ttaacacact atccttatta ctcaataaaa
     2701 ttattttttg ttttaacata tgatttatag aatctaaaaa ttgatattga cttataatag
     2761 tactttctac ctctctagct ttttctagtg tttcaaattc ttcaaactct gctatattgc
     2821 tatagtatag cttatcatta ataatattat ctgacaactt taattccatt ttataaaaca
     2881 attccaagtt atgtataaaa tttttaatgt tacaacatat agacttatta tttgttctta
     2941 tatggtatct ccatttacta cttctagctc tcatgttata tgcacaacta ctcccaacat
     3001 aaacatatat tgcttgttct aactcatatt tttgtaaaat ttcaacatca tatccagtca
     3061 aatcgtactc tctcttatat acaaatggtt tttctaattt tttgttcaat aaatctcctc
     3121 ctatttctta cttaatttag ctaaaaatct atttatattt cacctccttt ttgactttaa
     3181 gcctacgaat accatattta tgattataac atcttctatt tttcccaaat ccttacatct
     3241 cccctaattt ttagacttaa gggcgaattc tgcatcaagc tagcataaaa ataagaagcc
     3301 tgcatttgca ggcttcttat ttttatggta ccttaagacc cactttcaca tttaagttgt
     3361 ttttctaatc cgcatatgat caattcaagg ccgaataaga aggctggctc tgcaccttgg
     3421 tgatcaaata attcgatagc ttgtcgtaat aatggcggca tactatcagt agtaggtgtt
     3481 tccctttctt ctttagcgac ttgatgctct tgatcttcca atacgcaacc taaagtaaaa
     3541 tgccccacag cgctgagtgc atataatgca ttctctagtg aaaaaccttg ttggcataaa
     3601 aaggctaatt gattttcgag agtttcatac tgtttttctg taggccgtgt acctaaatgt
     3661 acttttgctc catcgcgatg acttagtaaa gcacatctaa aacttttagc gttattacgt
     3721 aaaaaatctt gccagctttc cccttctaaa gggcaaaagt gagtatggtg cctatctaac
     3781 atctcaatgg ctaaggcgtc gagcaaagcc cgcttatttt ttacatgcca atacaatgta
     3841 ggctgctcta cacctagctt ctgggcgagt ttacgggttg ttaaaccttc gattccgacc
     3901 tcattaagca gctctaatgc gctgttaatc actttacttt tatctaatct agacatcatt
     3961 aattcctcct ttttgttgac attatatcat tgatagagtt atttgtcaaa ctagtttttt
     4021 atttcgatgc cctggacttc atgaaaaact aaaaaaaata ttgacactct atcattgata
     4081 gagtataatt aaaataagct tgatcgtagc gttaacagat ctgagctcct gcagtaaagg
     4141 agaaaatttt atgagaccaa gtaaaaaatt attaatagct ataatatcaa tattcttaat
     4201 aagttcagtt ccagtaagtg cacatgcaga tagtgcaaac aaaagtaatg aattaacagc
     4261 acctagtatc aaaagtggaa caattctaca tgcttggaac tggtcattta acactttgaa
     4321 acacaatatg aaagatatac atgatgctgg atatacagca atacagactt ctccaataaa
     4381 ccaggtaaaa gagggaaatc aaggggataa atcaatgtct aattggtatt ggttatatca
     4441 accaacatca tatcaaatag gaaataggta tcttggtact gaacaagagt ttaaagaaat
     4501 gtgtgctgca gctgaagaat atggaataaa ggttattgta gatgctgtca taaatcatac
     4561 aacttcagac tatgctgcta tatctaacga agtaaagtct attccaaatt ggacacatgg
     4621 taatacacag attaagaatt ggtctgatcg atgggatgtg acacaaaact ctttactagg
     4681 tctttatgat tggaatactc aaaatactca agtacaatca tatcttaaaa gatttttgga
     4741 cagggctctt aacgatggag cagatggttt tagattcgat gcagcaaaac acatagaatt
     4801 gccagacgat ggatcatatg gtagtcagtt ttggccaaac ataacaaata ctagtgctga
     4861 atttcaatat ggtgagattt tacaagattc tgcaagtaga gatgcagcat atgcgaacta
     4921 tatggatgtt actgctagta attacggtca tagtattaga tcagctctaa agaataggaa
     4981 tttaggagtt tctaatatct ctcattatgc ctctgatgtt tcagcagata aactagtaac
     5041 ttgggttgaa agccatgata catatgcaaa cgacgatgaa gaatctacat ggatgtctga
     5101 tgatgatata agattaggtt gggcagttat agcatctaga agtggttcta caccattatt
     5161 cttttcaaga cctgaaggtg gaggaaatgg agtgcgattt ccaggaaaaa gtcaaattgg
     5221 tgatagaggt tcagccttat ttgaggacca agcaattact gctgtaaata gatttcataa
     5281 tgtaatggca gggcaacctg aagaattaag taatccaaat gggaacaatc aaatctttat
     5341 gaatcaacgt ggtagtcacg gagtagtttt agcaaatgca ggaagttcat ccgtttcaat
     5401 aaacactgca acaaagttac ctgatggtag atatgacaat aaagcaggag cagggagttt
     5461 tcaagtaaat gatgggaaac taacaggtac aattaacgca agatcagtag ctgttctata
     5521 tcctgatgat atcgcaaaag ccccacatgt attcttggag aattacaaaa caggtgtgac
     5581 gcattccttt aatgatcaat taactataac attaagagct gatgctaata caactaaagc
     5641 tgtctaccaa atcaataatg gtccagatga tagaagattg agaatggaga ttaacagtca
     5701 atcagaaaaa gaaattcaat ttggtaaaac atatacaata atgttaaagg gaactaactc
     5761 tgatggagta acaaggacgg aaaagtattc ttttgtcaaa agagatccag cttcagctaa
     5821 gacgatagga tatcagaacc ctaatcattg gtcacaagtg aatgcttata tttataaaca
     5881 tgatggatct agagttatag aattaaccgg atcatggcca ggtaaaccta tgactaaaaa
     5941 cgctgatgga atatacactt taacactacc agcagacaca gatactacta atgctaaagt
     6001 gatattcaat aacggaagtg cacaagtacc tggtcagaac caacctggtt ttgactacgt
     6061 acttaatgga ttatacaatg atagcggatt atctggaagt ttaccacatt aaggatccta
     6121 taagttttaa taaaacttta aatagaaaaa ggcttctctc atgagaagtc ttttttattt
     6181 aaaataaata taaaataaaa tagaggctat aaatagcctc tattttatgt gagaaatccc
     6241 taaataaaaa gatgccagtg tgctggaatt cgcccttagg ctgcgctagg gacctcttta
     6301 gctccttgga agctgtcagt agtaggggat cggtcttgcc ttgctcgtcg gtgatgtact
     6361 tcaccagctc cgcgaagtcg ctcttcttga tggagcgcat ggggacgtgc ttggcaatca
     6421 cgcgcacccc ccggccgttt tagcggctaa aaaagtcatg gctctgccct cgggcggacc
     6481 acgcccatca tgaccttgcc aagctcgtcc tgcttctctt cgatcttcgc cagcagggcg
     6541 aggatcgtgg catcaccgaa ccgcgccgtg cgcgggtcgt cggtgagcca gagtttcagc
     6601 aggccgccca ggcggcccag gtcgccattg atgcgggcca gctcgcggac gtgctcatag
     6661 tccacgacgc ccgtgatttt gtagccctgg ccgacggcca gcaggtaggc cgacaggctc
     6721 atgccggccg ccgccgcctt ttcctcaatc gctcttcgtt cgtctggaag gcagtacacc
     6781 ttgataggtg ggctgccctt cctggttggc ttggtttcat cagccatccg cttgccctca
     6841 tctgttacgc cggcggtagc cggccagcct cgcagagcag gattcccgtt gagcaccgcc
     6901 aggtgcgaat aagggacagt gaagaaggaa cacccgctcg cgggtgggcc tacttcacct
     6961 atcctgcccg gctgacgccg ttggatacac caaggaaagt ctacacgaac cctttggcaa
     7021 aatcctgtat atcgtgcgaa aaaggatgga tataccgaaa aaatcgctat aatgaccccg
     7081 aagcaggaag ggcgaattct gcagatctga ccggtctctg aaaatataaa aaccacagat
     7141 tgatactaaa accttggttg tgttgctttt cggggcttaa atcaaggaaa aatccttgtt
     7201 ttaagccttt caaaaagaaa cacaaggtct ttgtactaac ctgtggttat gtataaaatt
     7261 gtagatttta gggtaacaaa aaacaccgta tttctacgat gtttttgctt aaatacttgt
     7321 ttttagttac agacaaacct gaagttaact atttatcaat tcctgcaatt cgtttacaaa
     7381 acggcaaatg tgaaatccgt cacatactgc gtgatgaact tgaattgcca aaggaagtat
     7441 aattttgtta tcttctttat aatatttccc catagtaaaa ataggaatca aataatcata
     7501 tcctttctgc aaattcagat taaagccatc gaaggttgac cacggtatca tagatacatt
     7561 aaaaatgttt tccggagcat ttggctttcc ttccattcta tgattgtttc cataccgttg
     7621 cgtatcactt tcataatctg ctaaaaatga tttaaagtca gacttacact cagtccaaag
     7681 gctggaaaat gtttcagtat cattgtgaaa tattgtatag cttggtatca tctcatcata
     7741 tatccccaat tcaccatctt gattgattgc cgtcctaaac tctgaatggc ggtttacaat
     7801 cattgcaata taataaagca ttgcaggata tagtttcatt cccttttcct ttatttgtgt
     7861 gatatccact ttaacggtca tgctgtaggt acaaggtaca cttgcaaagt agtggtcaaa
     7921 atactctttt ctgttccaac tatttttatc aattttttca aataccatct aagttccctc
     7981 tcaaattcaa gtttatcgct ctaatgaaca aagatattat accacatttt tgtgaatttt
     8041 tcaacttgcc cacttcgact gcactcccga cttaataact tcttgaacac ttgccgaaaa
     8101 agaaaaactg ccgggtacgt acccgggatc gatccccgcc gagcgctctt ccgcttcctc
     8161 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     8221 ggcggtaata cggttatcca ctgaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     8281 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     8341 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     8401 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     8461 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     8521 tcaatgctca cgctgtaggt atcacagttc ggtgtaggtc gttcgctcca agctgggctg
     8581 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     8641 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     8701 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     8761 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     8821 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     8881 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     8941 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgaaatgcaa
     9001 gtttctaact aacatacatc atagttacta aactatggta actataattt tattaactat
     9061 attaagcact gattagtact ataactcaat ataagcatat cccctgtatc gtaactagag
     9121 aaccaaacga cggaaaaagc gatatagata agtttagata
//
