LOCUS       sequence-123367-        6537 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 6537)
  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 6537)
  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: MP1
FEATURES             Location/Qualifiers
     source          1..6537
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             2484..3545
                     /codon_start=1
                     /gene="recA"
                     /product="DNA repair protein from E. coli"
                     /label=RecA
                     /translation="MAIDENKQKALAAALGQIEKQFGKGSIMRLGEDRSMDVKTISTGS
                     LSLDIALGAGGLPMGRIVEIYGPESSGKTTLTLQVIAAAQREGKTCAFIDAEHALDPIY
                     ARKLGVDIDNLLCSQPDTGEQALEICDALARSGAVDVIVVDSVAALTPKAEIEGEIGDS
                     HMGLAARMMSQAMRKLAGNLKQSNTLLIFINQIRMKIGVMFGNPETTTGGNALKFYASV
                     RLDIRRIGAVKEGENVVGSETRVKVVKNKIAAPFKQAEFQILYGEGINFYGELVDLGVK
                     EKLIEKAGAWYSYKGEKIGQGKANATAWLKDNPETAKEIEKKVRELLLSNPNSTPDFSV
                     DDSEGVAETNEDF"
     terminator      3572..3603
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3619..4278)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4193..4212
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4431..5169)
                     /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      5259..5304
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5319..6197)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        6224..6508
                     /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     6431..6450
                     /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 aaaccggtat gcctcacaca ggaaacagaa
      781 ttcattacca tgggctttcc ttcaccggca gcagattacg ttgaacagcg catcgatctg
      841 aatcaactgt tgatccagca tcccagcgcg acttacttcg tcaaagcaag tggtgattct
      901 atgattgatg gtggaattag tgacggtgat ttactgattg tcgatagcgc tattaccgcc
      961 agccatggtg atattgtcat cgctgctgtt gacggcgagt ttacggtgaa aaaattgcaa
     1021 ctacgcccga cggtacagct tattcccatg aacagcgcgt actcgcccat taccatcagt
     1081 agtgaagata cgctggatgt ctttggtgtg gtgatccacg tcgttaaggc gatgcgctga
     1141 tgtttgccct ctgtgatgta aacgcgtttt atgccagctg tgagacggtg tttcgccctg
     1201 atttatgggg taaaccggtg gttgtgctat cgaataatga cggttgcgtt atcgcccgaa
     1261 acgctgaggc aaaggcgctt ggcgttaaaa tgggcgatcc ctggttcaaa caaaaagatc
     1321 tgtttcgtcg ctgtggcgtg gtttgcttta gcagcaatta tgagctttac gcagacatga
     1381 gcaatcgggt gatgtcgacg ctggaagagc tatcgccccg cgtcgagatt tacagtaccg
     1441 gtatgcctat tgatgaggca ttctgcgatc tgacaggtgt gcgtaattgt cgcgatctga
     1501 ctgattttgg cagagaaatt cgcgcaacgg tgctacaacg tacccatctt actgttggtg
     1561 tggggatcgc ccagaccaaa acgctggcta agcttgccaa tcatgcggca aaaaaatggc
     1621 agcggcagac gggtggggtg gtggatttat caaatctgga acgccagcgt aaattaatgt
     1681 ctgctctccc cgtggatgac gtctggggga ttggacggcg gatcagcaaa aaactggacg
     1741 cgatggggat caaaaccgtt ctcgatttgg cggatacaga tatccggttt atccgtaaac
     1801 attttaatgt cgtgctcgaa agaacggtgc gtgaactgcg cggcgaaccc tgtttgcaac
     1861 tggaagagtt tgcaccgacg aagcaggaaa ttatctgttc ccgctcgttt ggtgaacgca
     1921 tcacggatta tccgtcgatg cggcaggcca tttgtagtta cgctgcccgg gcggcggaaa
     1981 aacttcgcag cgagcatcaa tattgtcgat taatttccac gtttattaag acgtcaccat
     2041 ttgcgctcaa tgaaccttat tacggcaata gcgcgtcggt aaaactgctg acgcccactc
     2101 aggacagcag ggatatcatt aacgctgcta cgcgatctct ggatgccatc tggcaagcgg
     2161 gccatcgtta ccaaaaagcg ggcgtgatgc tgggggattt cttcagtcag ggagtcgcgc
     2221 agctcaattt attcgatgac aacgcaccgc gccccgggag tgagcaattg atgacggtaa
     2281 tggatacact gaatgctaaa gagggcagag gaacactcta ttttgccggg caggggatcc
     2341 agcaacaatg gcagatgaag cgagccatgc tttcaccacg ttatacaacg cgaagttctg
     2401 atttactgag ggtcaaataa atatagcggc aggaaaaaag cgatcccgca tatccggtat
     2461 tacccggcat gacaggagta aaaatggcta tcgacgaaaa caaacagaaa gcgttggcgg
     2521 cagcactggg ccagattgag aaacaatttg gtaaaggctc catcatgcgc ctgggtgaag
     2581 accgttccat ggatgtgaaa accatctcta ccggttcgct ttcactggat atcgcgcttg
     2641 gggcaggtgg tctgccgatg ggccgtatcg tcgaaatcta cggaccggaa tcttccggta
     2701 aaaccacgct gacgctgcag gtgatcgccg cagcgcagcg tgaaggtaaa acctgtgcgt
     2761 ttatcgatgc tgaacacgcg ctggacccaa tctacgcacg taaactgggc gtcgatatcg
     2821 ataacctgct gtgctcccag ccggacaccg gcgagcaggc actggaaatc tgtgacgccc
     2881 tggcgcgttc tggcgcagta gacgttatcg tcgttgactc cgtggcggca ctgacgccga
     2941 aagcggaaat cgaaggcgaa atcggcgact ctcacatggg ccttgcggca cgtatgatga
     3001 gccaggcgat gcgtaagctg gcgggtaacc tgaagcagtc caacacgctg ctgatcttca
     3061 tcaaccagat ccgtatgaaa attggtgtga tgttcggtaa cccggaaacc actaccggtg
     3121 gtaacgcgct gaaattctac gcctctgttc gtctcgacat ccgtcgtatc ggcgcggtga
     3181 aagagggcga aaacgtggtg ggtagcgaaa cccgcgtgaa agtggtgaag aacaaaatcg
     3241 ctgcgccgtt taaacaggct gaattccaga tcctctacgg cgaaggtatc aacttctacg
     3301 gcgaactggt tgacctgggc gtaaaagaga agctgatcga gaaagcaggc gcgtggtaca
     3361 gctacaaagg tgagaagatc ggtcagggta aagcgaatgc gactgcctgg ctgaaagata
     3421 acccggaaac cgcgaaagag atcgagaaga aagtacgtga gttgctgctg agcaacccga
     3481 actcaacgcc ggatttctct gtagatgata gcgaaggcgt agcagaaact aacgaagatt
     3541 tttaaactta attaacggca ctcctcagcc aagtcaaaag cctccgaccg gaggcttttg
     3601 actacatgcc catggcgttt acgccccgcc ctgccactca tcgcagtact gttgtaattc
     3661 attaagcatt ctgccgacat ggaagccatc acaaacggca tgatgaacct gaatcgccag
     3721 cggcatcagc accttgtcgc cttgcgtata atatttgccc atagtgaaaa cgggggcgaa
     3781 gaagttgtcc atattggcca cgtttaaatc aaaactggtg aaactcaccc agggattggc
     3841 tgagacgaaa aacatattct caataaaccc tttagggaaa taggccaggt tttcaccgta
     3901 acacgccaca tcttgcgaat atatgtgtag aaactgccgg aaatcgtcgt ggtattcact
     3961 ccagagcgat gaaaacgttt cagtttgctc atggaaaacg gtgtaacaag ggtgaacact
     4021 atcccatatc accagctcac cgtctttcat tgccatacgg aactccggat gagcattcat
     4081 caggcgggca agaatgtgaa taaaggccgg ataaaacttg tgcttatttt tctttacggt
     4141 ctttaaaaag gccgtaatat ccagctgaac ggtctggtta taggtacatt gagcaactga
     4201 ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat atatcaacgg tggtatatcc
     4261 agtgattttt ttctccattt tagcttcctt agctcctgaa aatctcgata actcaaaaaa
     4321 tacgcccggt agtgatctta tttcattatg gtgaaagttg gaacctctta cgtgccaagc
     4381 caaataggcc gtcactcggt cgctacgctc cgggcgtgag actgcggcgg gcgctgcgga
     4441 cacatacaaa gttacccaca gattccgtgg ataagcaggg gactaacatg tgaggcaaaa
     4501 cagcagggcc gcgccggtgg cgtttttcca taggctccgc cctcctgcca gagttcacat
     4561 aaacagacgc ttttccggtg catctgtggg agccgtgagg ctcaaccatg aatctgacag
     4621 tacgggcgaa acccgacagg acttaaagat ccccaccgtt tccggcgggt cgctccctct
     4681 tgcgctctcc tgttccgacc ctgccgttta ccggatacct gttccgcctt tctcccttac
     4741 gggaagtgtg gcgctttctc atagctcaca cactggtatc tcggctcggt gtaggtcgtt
     4801 cgctccaagc tgggctgtaa gcaagaactc cccgttcagc ccgactgctg cgccttatcc
     4861 ggtaactgtt cacttgagtc caacccggaa aagcacggta aaacgccact ggcagcagcc
     4921 attggtaact gggagttcgc agaggatttg tttagctaaa cacgcggttg ctcttgaagt
     4981 gtgcgccaaa gtccggctac actggaagga cagatttggt tgctgtgctc tgcgaaagcc
     5041 agttaccacg gttaagcagt tccccaactg acttaacctt cgatcaaacc acctccccag
     5101 gtggtttttt cgtttacagg gcaaaagatt acgcgcagaa aaaaaggatc tcaagaagat
     5161 cctttgatct tttctactga accgctctag atttcagtgc aatttatctc ttcaaatgta
     5221 gcacctgaag tcagcccagg aggaagagga catccggtca aataaaacga aaggctcagt
     5281 cgaaagactg ggcctttcgt tttagactta gggacccttt atgacaactt gacggctaca
     5341 tcattcactt tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt
     5401 ttaaataccc gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg
     5461 ataggcatcc gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc
     5521 cagcttaaga cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac
     5581 aagcaaacat gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg
     5641 atgtactgac aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc
     5701 gcttccatgc gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag
     5761 cgcccttccc cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg
     5821 tgcgcttcat ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat
     5881 tcatgccagt aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc
     5941 ggatgacgac cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg
     6001 caaacaaatt ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga
     6061 atataacctt tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca
     6121 atcggcgtta aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca
     6181 ttttgcgctt cagccatact tttcatactc ccaccattca gagaagaaac caattgtcca
     6241 tattgcatca gacattgccg tcactgcgtc ttttactggc tcttctcgct aacccaaccg
     6301 gtaaccccgc ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg
     6361 cgtaacaaaa gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc
     6421 acactttgct atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt
     6481 ttatcgcaac tctctactgt ttctccatac ccgttttttt ggacgcgtac aactcaa
//
