LOCUS       sequence-330952-        9357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MazF-based mutagenesis vector for C. difficile.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9357)
  AUTHORS   Fuchs M, Lamm-Schmidt V, Sulzer J, Ponath F, Jenniches L, Kirk JA, 
            Fagan RP, Barquist L, Vogel J, Faber F
  TITLE     An RNA-centric global view of Clostridioides difficile reveals broad
            activity of Hfq in a clinically important gram-positive bacterium.
  JOURNAL   Proc Natl Acad Sci U S A. 2021 Jun 22;118(25). pii: 2103579118. doi:
            10.1073/pnas.2103579118.
  PUBMED    34131082
REFERENCE   2  (bases 1 to 9357)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2021-06-22"; volume: "118(25). pii: 
            2103579118. doi"; pages: " 10.1073/pnas.2103579118"
COMMENT     Sequence Label: pJAK184
FEATURES             Location/Qualifiers
     source          1..9357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     363..382
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     616..633
                     /label=L4440
                     /note="L4440 vector, forward primer"
     oriT            783..892
                     /note="incP origin of transfer"
     CDS             925..1296
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     primer_bind     1493..1509
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1493..1509
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3344..3361)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3344..3360)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3353..3375)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(9231..9357,1..462)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
       61 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
      121 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
      181 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
      241 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
      301 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
      361 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      421 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      481 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      541 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      601 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcagg
      661 gccccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
      721 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
      781 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
      841 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc
      901 ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc
      961 ccacctatca aggtgtactg ccttccagac gaacgaagag cgattgagga aaaggcggcg
     1021 gcggccggca tgagcctgtc ggcctacctg ctggccgtcg gccagggcta caaaatcacg
     1081 ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca tcaatggcga cctgggccgc
     1141 ctgggcggcc tgctgaaact ctggctcacc gacgacccgc gcacggcgcg gttcggtgat
     1201 gccacgatcc tcgccctgct ggcgaagatc gaagagaagc aggacgagct tggcaaggtc
     1261 atgatgggcg tggtccgccc gagggcagag ccatgacttt tttagccgct aaaacggccg
     1321 gggggtgcgc gtgattgcca agcacgtccc catgcgctcc atcaagaaga gcgacttcgc
     1381 ggagctggtg aagtacatca ccgacgagca aggcaagacc gatcgggccc cctgcaggat
     1441 aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt atcaggaaac
     1501 agctatgacc gcggccgcgc tgtatatagt cataaccctt atatttcatt aattaaagtt
     1561 aaattttgta tattcaaaaa gctcttgata tttacggcag agccaccata tagagatgcc
     1621 tcagacccta atgatgaaat ttctatattt attcctttgt taaaagagct aactaacata
     1681 tcttttgtaa tctgtaatat ctcagggata tcactaatta tttggctatt taaatatatt
     1741 atttcaggag cataagttgt aatagcatta tttatagcta ttgttaagta actacaaaac
     1801 tcatgaataa cttttttggc attctggtta tcttcataat atagttgctt tactatatca
     1861 gaatctattt taggaatatt ttctaaagaa gaaagctgtt caaatacctt tttttctgaa
     1921 caatactgtt ctaaacagcc acgatttcca caaggacata gtttaccatt aggcatgatg
     1981 atagtgtgac caatttctcc actcatacca tttctgccgc tatataattt attatttatt
     2041 ataatgcctg aaccaaatcc actgtgaata ctgagactaa gtagtgagtt atgtacagtt
     2101 gaaaaagtat tctcagctaa agctgttaag tttgcttcat tttctatatg aataggaaag
     2161 tcatactttt tgctaaggat gctatataag tcaatctcat taagattata atatggagta
     2221 aacaatactt tattttcaca agtaatccca tgtatagcca aagtaagacc aattacctta
     2281 taaggagtgt ctattttgga tatgttataa ctatttataa tttcatctat caattgtata
     2341 acattgtctt tacttacttg tatatctgtt aatttcttag aatttataat agttccatct
     2401 aagtaagata gagaagaaaa tatatagtcg tatccaatgt ccatacttaa agaaatccct
     2461 gcacatttat taaatactaa taatatgggt tttcttccgc cactatgagt actatttcca
     2521 attcctattt catgaactag agattcatca ataagttttt tggtaatagc ggatatagtt
     2581 gctttattca atcctatagt agaagctata cttgccctag aaataggacc attttttata
     2641 atttgttcaa gtaccaatct ttcattcatt tctcgaatag tgtatttatc agtaaccaat
     2701 ttgatattcc tccttaaaat aatattgtaa tactttttac acaaaaataa aaggttattt
     2761 tgcattgaca aagataatta aatattttat tattagttca taagttagtt taatatacta
     2821 acaaaaataa agcaagtaaa atatacctaa aatataaaaa aattaggata ggaaaacgat
     2881 agttatgaag tggcgcagta aaggagaaaa ttttatggta aggaaaacga tagttatgaa
     2941 gtggcgcagt aaaggagaaa attttatggt aagtagatat gtaccagata tgggagattt
     3001 aatatgggtt gattttgatc caacaaaagg tagtgaacaa gcaggacata gaccagctgt
     3061 tgtattaagt ccatttatgt ataataataa aacaggtatg tgtttatgtg ttccttgtac
     3121 aactcaatca aaaggatatc catttgaagt tgttttatca ggtcaagaaa gagatggagt
     3181 agcattagct gatcaagtaa aaagtatagc atggagagca agaggagcaa ctaaaaaagg
     3241 aacagttgca ccagaagaat tacaattaat taaagcaaaa attaatgtat taattggata
     3301 aatcaacaac tctcctcgag gcctgcagac atgcaagctt ggcactggcc gtcgttttac
     3361 aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc
     3421 ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc
     3481 gcagcctgaa tggcgaatgg cgctagcata aaaataagaa gcctgcattt gcaggcttct
     3541 tatttttatg gcgcgccgtt ctgaatcctt agctaatggt tcaacaggta actatgacga
     3601 agatagcacc ctggataagt ctgtaatgga ttctaaggca tttaatgaag acgtgtatat
     3661 aaaatgtgct aatgaaaaag aaaatgcgtt aaaagagcct aaaatgagtt caaatggttt
     3721 tgaaattgat tggtagttta atttaatata ttttttctat tggctatctc gatacctata
     3781 gaatcttctg ttcacttttg tttttgaaat ataaaaaggg gctttttagc cccttttttt
     3841 taaaactccg gaggagtttc ttcattcttg atactatacg taactatttc gatttgactt
     3901 cattgtcaat taagctagta aaatcaatgg ttaaaaaaca aaaaacttgc atttttctac
     3961 ctagtaattt ataattttaa gtgtcgagtt taaaagtata atttaccagg aaaggagcaa
     4021 gttttttaat aaggaaaaat ttttcctttt aaaattctat ttcgttatat gactaattat
     4081 aatcaaaaaa atgaaaataa acaagaggta aaaactgctt tagagaaatg tactgataaa
     4141 aaaagaaaaa atcctagatt tacgtcatac atagcacctt taactactaa gaaaaatatt
     4201 gaaaggactt ccacttgtgg agattatttg tttatgttga gtgatgcaga cttagaacat
     4261 tttaaattac ataaaggtaa tttttgcggt aatagatttt gtccaatgtg tagttggcga
     4321 cttgcttgta aggatagttt agaaatatct attcttatgg agcatttaag aaaagaagaa
     4381 aataaagagt ttatattttt aactcttaca actccaaatg taaaaagtta tgatcttaat
     4441 tattctatta aacaatataa taaatctttt aaaaaattaa tggagcgtaa ggaagttaag
     4501 gatataacta aaggttatat aagaaaatta gaagtaactt accaaaagga aaaatacata
     4561 acaaaggatt tatggaaaat aaaaaaagat tattatcaaa aaaaaggact tgaaattggt
     4621 gatttagaac ctaattttga tacttataat cctcattttc atgtagttat tgcagttaat
     4681 aaaagttatt ttacagataa aaattattat ataaatcgag aaagatggtt ggaattatgg
     4741 aagtttgcta ctaaggatga ttctataact caagttgatg ttagaaaagc aaaaattaat
     4801 gattataaag aggtttacga acttgcgaaa tattcagcta aagacactga ttatttaata
     4861 tcgaggccag tatttgaaat tttttataaa gcattaaaag gcaagcaggt attagttttt
     4921 agtggatttt ttaaagatgc acacaaattg tacaagcaag gaaaacttga tgtttataaa
     4981 aagaaagatg aaattaaata tgtctatata gtttattata attggtgcaa aaaacaatat
     5041 gaaaaaacta gaataaggga acttacggaa gatgaaaaag aagaattaaa tcaagattta
     5101 atagatggaa tagaaataga ttaaagtgta actatacttt atatatatat gattaaaaaa
     5161 ataaaaaaca acagcctatt aggttgttgt tttttatttt ctttattaat ttttttaatt
     5221 tttagttttt agttcttttt taaaataagt ttcagcctct ttttcaatat tttttaaaga
     5281 aggagtattt gcatgaattg ccttttttct aacagactta ggaaatattt taacagtatc
     5341 ttcttgcgcc ggtgattttg gaacttcata acttactaat ttataattat tattttcttt
     5401 tttaattgta acagttgcaa aagaagctga acctgttcct tcaactagtt tatcatcttc
     5461 aatataatat tcttgaccta tatagtataa atatattttt attatatttt tacttttttc
     5521 tgaatctatt attttataat cataaaaagt tttaccacca aaagaaggtt gtactccttc
     5581 tggtccaaca tattttttta ctatattatc taaataattt ttgggaactg gtgttgtaat
     5641 ttgattaatc gaacaaccag ttatacttaa aggaattata actataaaaa tatataggat
     5701 tatcttttta aatttcatta ttggcctcct ttttattaaa tttatgttac cataaaaagg
     5761 acataacggg aatatgtaga atatttttaa tgtagacaaa attttacata aatataaaga
     5821 aaggaagtgt ttgtttaaat tttatagcaa actatcaaaa attaggggga taaaaattta
     5881 tgaaaaaaag gttttcgatg ttatttttat gtttaacttt aatagtttgt ggtttattta
     5941 caaattcggc cggccagtgg gcaagttgaa aaattcacaa aaatgtggta taatatcttt
     6001 gttcattaga gcgataaact tgaatttgag agggaactta gatggtattt gaaaaaattg
     6061 ataaaaatag ttggaacaga aaagagtatt ttgaccacta ctttgcaagt gtaccttgta
     6121 cctacagcat gaccgttaaa gtggatatca cacaaataaa ggaaaaggga atgaaactat
     6181 atcctgcaat gctttattat attgcaatga ttgtaaaccg ccattcagag tttaggacgg
     6241 caatcaatca agatggtgaa ttggggatat atgatgagat gataccaagc tatacaatat
     6301 ttcacaatga tactgaaaca ttttccagcc tttggactga gtgtaagtct gactttaaat
     6361 catttttagc agattatgaa agtgatacgc aacggtatgg aaacaatcat agaatggaag
     6421 gaaagccaaa tgctccggaa aacattttta atgtatctat gataccgtgg tcaaccttcg
     6481 atggctttaa tctgaatttg cagaaaggat atgattattt gattcctatt tttactatgg
     6541 ggaaatatta taaagaagat aacaaaatta tacttccttt ggcaattcaa gttcatcacg
     6601 cagtatgtga cggatttcac atttgccgtt ttgtaaacga attgcaggaa ttgataaata
     6661 gttaacttca ggtttgtctg taactaaaaa caagtattta agcaaaaaca tcgtagaaat
     6721 acggtgtttt ttgttaccct agagctctat acaggtgaag cagatgctat gagtgcgtca
     6781 tcagtagcat caagagatgg atcacctata attttaacta atggtaaaaa tgttccattt
     6841 gagaaaaaag aaggtgtgca atgttatgct cttggttcag aggagataat aagtaatgat
     6901 ttagttaaaa aaactaatgc agttagatta gctggagaag atagatttga gactaataaa
     6961 aaagttataa aacattttta tagttcggca aaagaatttt atctttctaa aggttatcaa
     7021 ttagtagatg cagttgctgg ttcttctata gcaaaaaatg cacctattgt attagttgac
     7081 ggaaatagtg ataagtctgt acttagaagt gctgataaga ttacagcact aggaggaata
     7141 gatgaaaaga cactagaaca atgtttaagt gcatcatcac tggatgcaag tgcaccaaca
     7201 ataactgttg gaaatttaaa tatttatcaa ggtgataagt ttgatataag taaactgaat
     7261 atcgtagcaa aggatagcaa tggtaacgat ttgacaccag agcttatagg aaatattgat
     7321 actgataagg tgggtaaata caaagttact ataaaagcta ctgatattgg tggaaagact
     7381 acatcaataa gtgtagaagt aaatgtactt gaatataaga ctaatgatat gaatagtagt
     7441 gagtttaaaa gaatggtttc tagtgaaatg tatagtttgg taaattcata tagaaaagaa
     7501 aaaggtaaag agccattaca agtttctaaa aacttagaag gactttctaa ttcctggtct
     7561 aagtacatgg cagataaagg tcagttttca catgtaataa atggtaagaa atctgttgaa
     7621 gtatttactg gatatggctt gagaagtgaa gaaaatatag cttttgtacc acttgttaca
     7681 aaaagtactt atacaacaaa ggatgctaga gaagtagcaa atgtaatatt tacagtttgg
     7741 aaaaagtcag ataaatataa tgaaaatatg ttaaatagtg attttgtata tactgggttt
     7801 ggattatata tattgcctaa tggagaagtg tatgctacac aagaattttt aaataagtaa
     7861 tttaaataag taaaatcaaa ttttttaagt tattaaatat tggagtttaa gggtataaaa
     7921 actcatatgc ttgggtataa atataataat ataaaaaggc ttcttatatg agaagtcttt
     7981 ttatattatg ttttcgaata caaaaaaaga gactaaattt agtctctttt ttatgtgaga
     8041 aatacttaaa tatataaaga tgtaatttat ttaagtatga ttattatact atacaataaa
     8101 ataattgtct actattctat aattttaagg tataataata attgtatgat aaaagaggag
     8161 gtcaaaaaat atgtcagttt tagatgcaat cttagataaa gcggatgaac aagaaattaa
     8221 aaaattaaat ttaatagtag ataaaataga atctttagaa aaagatatgg aaatcaaaga
     8281 gaatgaacaa ttaaaagaaa tgacaaatac ttttagactt agattagaca aaggtgaaag
     8341 tttggatgat atacttccag aagcatttgc tgttgttaga gaggtctcaa aaagagtttt
     8401 aggaatgcgt caatataaag ttcagttaat tggaggtata gtaattcatc aaggaaagat
     8461 agctgagatg aagactggtg aggggaaaac tttagttgag gtagctccag tatatctaaa
     8521 tgctttaaca ggaaaaggtg tacatgtaat tacagtaaat gattatttgg cagaacgtga
     8581 taaagaactt atgcgtccag tatatgaatt tttaggattg actgtaggtg taattttatc
     8641 taaccaagac ccaagtataa gaaaagaaca atataaatgt gatataactt atggaacaaa
     8701 tagtgaattt ggatttgact atttaagaga taatatggtg ccagatttgt ctcataaagt
     8761 acaaagggaa ctaaactttg ccatagtaga tgagatagac tcaatattaa tagatgaagc
     8821 tagaactcca cttattattg caggagatgg tgatgaagat ttaaaacttt atgagttagc
     8881 aaataacttt gtaaaaactg ttaaagaaga agactttgag atggatagaa aagataaaac
     8941 tatagcatta acagcaagtg gtataagcaa agctgagtca ttttttggta taacaaacct
     9001 tactgatata aagaatatag aattatatca tcatataaat caagctttaa gaggtcataa
     9061 gcttatggaa aaagatgttg actatgttat ctcaaatgga gaagtaatga tagttgacga
     9121 atttacagga agagggatcc cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     9181 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     9241 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     9301 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcaga
//
