LOCUS       sequence-123368-        3744 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Untargetted Bacterial Mutagenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3744)
  AUTHORS   Badran AH, Liu DR
  TITLE     Development of potent in vivo mutagenesis plasmids with broad 
            mutational spectra.
  JOURNAL   Nat Commun. 2015 Oct 7;6:8425. doi: 10.1038/ncomms9425.
  PUBMED    26443021
REFERENCE   2  (bases 1 to 3744)
  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: "Nat 
            Commun."; date: "2015-10-7"; volume: "6:8425. doi"; pages: " 
            10.1038/ncomms9425"
COMMENT     Sequence Label: MP2
FEATURES             Location/Qualifiers
     source          1..3744
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      779..810
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(826..1485)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1400..1419
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(1638..2376)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     terminator      2466..2511
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2526..3404)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        3431..3715
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     3638..3657
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 gtctgacata aatgaccgct atgagcactg caattacacg ccagatcgtt ctcgctaccg
       61 caaccaccgg tatgaaccag attggtgcgc actatgaagg ccacaagatc attgagattg
      121 gtgccgttga agtggtgaac cgtcgcctga cgggcaataa cttccatgtt tatctcaaac
      181 ccgatcggct ggtggatccg gaagcctttg gcgtacatgg tattgccgat gaatttttgc
      241 tcgataagcc cacgtttgcc gaagtagccg atgagttcat ggactatatt cgcggcgcgg
      301 agttggtgat ccataacgca gcgttcgata tcggctttat ggactacgag ttttcgttgc
      361 ttaagcgcga tattccgaag accaatactt tctgtaaggt caccgatagc cttgcggtgg
      421 cgaggaaaat gtttcccggt aagcgcaaca gcctcgatgc gttatgtgct cgctacgaaa
      481 tagataacag taaacgaacg ctgcacgggg cattactcga tgcccagatc cttgcggaag
      541 tttatctggc gatgaccggt ggtcaaacgt cgatggcttt tgcgatggaa ggagagacac
      601 aacagcaaca aggtgaagca acaattcagc gcattgtacg tcaggcaagt aagttacgcg
      661 ttgtttttgc gacagatgaa gagattgcag ctcatgaagc ccgtctcgat ctggtgcaga
      721 agaaaggcgg aagttgcctc tggcgagcat aaacttaatt aacggcactc ctcagccaag
      781 tcaaaagcct ccgaccggag gcttttgact acatgcccat ggcgtttacg ccccgccctg
      841 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
      901 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
      961 tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
     1021 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
     1081 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
     1141 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
     1201 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
     1261 catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
     1321 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
     1381 ctggttatag gtacattgag caactgactg aaatgcctca aaatgttctt tacgatgcca
     1441 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
     1501 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
     1561 aaagttggaa cctcttacgt gccaagccaa ataggccgtc actcggtcgc tacgctccgg
     1621 gcgtgagact gcggcgggcg ctgcggacac atacaaagtt acccacagat tccgtggata
     1681 agcaggggac taacatgtga ggcaaaacag cagggccgcg ccggtggcgt ttttccatag
     1741 gctccgccct cctgccagag ttcacataaa cagacgcttt tccggtgcat ctgtgggagc
     1801 cgtgaggctc aaccatgaat ctgacagtac gggcgaaacc cgacaggact taaagatccc
     1861 caccgtttcc ggcgggtcgc tccctcttgc gctctcctgt tccgaccctg ccgtttaccg
     1921 gatacctgtt ccgcctttct cccttacggg aagtgtggcg ctttctcata gctcacacac
     1981 tggtatctcg gctcggtgta ggtcgttcgc tccaagctgg gctgtaagca agaactcccc
     2041 gttcagcccg actgctgcgc cttatccggt aactgttcac ttgagtccaa cccggaaaag
     2101 cacggtaaaa cgccactggc agcagccatt ggtaactggg agttcgcaga ggatttgttt
     2161 agctaaacac gcggttgctc ttgaagtgtg cgccaaagtc cggctacact ggaaggacag
     2221 atttggttgc tgtgctctgc gaaagccagt taccacggtt aagcagttcc ccaactgact
     2281 taaccttcga tcaaaccacc tccccaggtg gttttttcgt ttacagggca aaagattacg
     2341 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctactgaacc gctctagatt
     2401 tcagtgcaat ttatctcttc aaatgtagca cctgaagtca gcccaggagg aagaggacat
     2461 ccggtcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt agacttaggg
     2521 accctttatg acaacttgac ggctacatca ttcacttttt cttcacaacc ggcacggaac
     2581 tcgctcgggc tggccccggt gcatttttta aatacccgcg agaaatagag ttgatcgtca
     2641 aaaccaacat tgcgaccgac ggtggcgata ggcatccggg tggtgctcaa aagcagcttc
     2701 gcctggctga tacgttggtc ctcgcgccag cttaagacgc taatccctaa ctgctggcgg
     2761 aaaagatgtg acagacgcga cggcgacaag caaacatgct gtgcgacgct ggcgatatca
     2821 aaattgctgt ctgccaggtg atcgctgatg tactgacaag cctcgcgtac ccgattatcc
     2881 atcggtggat ggagcgactc gttaatcgct tccatgcgcc gcagtaacaa ttgctcaagc
     2941 agatttatcg ccagcagctc cgaatagcgc ccttcccctt gcccggcgtt aatgatttgc
     3001 ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg ggcgaaagaa ccccgtattg
     3061 gcaaatattg acggccagtt aagccattca tgccagtagg cgcgcggacg aaagtaaacc
     3121 cactggtgat accattcgcg agcctccgga tgacgaccgt agtgatgaat ctctcctggc
     3181 gggaacagca aaatatcacc cggtcggcaa acaaattctc gtccctgatt tttcaccacc
     3241 ccctgaccgc gaatggtgag attgagaata taacctttca ttcccagcgg tcggtcgata
     3301 aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac ccgccaccag atgggcatta
     3361 aacgagtatc ccggcagcag gggatcattt tgcgcttcag ccatactttt catactccca
     3421 ccattcagag aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt
     3481 tactggctct tctcgctaac ccaaccggta accccgctta ttaaaagcat tctgtaacaa
     3541 agcgggacca aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa
     3601 gtccacattg attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata
     3661 agattagcgg atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg
     3721 tttttttgga cgcgtacaac tcaa
//
