LOCUS       sequence-123376-        5705 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 5705)
  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 5705)
  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: MP-E
FEATURES             Location/Qualifiers
     source          1..5705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(39..777)
                     /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      867..912
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(927..1805)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1832..2116
                     /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     2039..2058
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     terminator      4885..4916
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4932..5591)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5506..5525
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
       61 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
      121 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
      181 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
      241 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
      301 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
      361 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
      421 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
      481 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
      541 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
      601 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
      661 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
      721 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
      781 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
      841 agcccaggag gaagaggaca tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg
      901 cctttcgttt tagacttagg gaccctttat gacaacttga cggctacatc attcactttt
      961 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
     1021 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
     1081 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
     1141 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
     1201 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
     1261 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
     1321 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
     1381 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
     1441 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
     1501 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
     1561 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
     1621 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
     1681 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
     1741 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
     1801 gccatacttt tcatactccc accattcaga gaagaaacca attgtccata ttgcatcaga
     1861 cattgccgtc actgcgtctt ttactggctc ttctcgctaa cccaaccggt aaccccgctt
     1921 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
     1981 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
     2041 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
     2101 tctactgttt ctccataccc gtttttttgg acgcgtacaa ctcaagtctg acataaatga
     2161 ccgctatgag cactgcaatt acacgccaga tcgttctcgc taccgcaacc accggtatga
     2221 accagattgg tgcgcactat gaaggccaca agatcattga gattggtgcc gttgaagtgg
     2281 tgaaccgtcg cctgacgggc aataacttcc atgtttatct caaacccgat cggctggtgg
     2341 atccggaagc ctttggcgta catggtattg ccgatgaatt tttgctcgat aagcccacgt
     2401 ttgccgaagt agccgatgag ttcatggact atattcgcgg cgcggagttg gtgatccata
     2461 acgcagcgtt cgatatcggc tttatggact acgagttttc gttgcttaag cgcgatattc
     2521 cgaagaccaa tactttctgt aaggtcaccg atagccttgc ggtggcgagg aaaatgtttc
     2581 ccggtaagcg caacagcctc gatgcgttat gtgctcgcta cgaaatagat aacagtaaac
     2641 gaacgctgca cggggcatta ctcgatgccc agatccttgc ggaagtttat ctggcgatga
     2701 ccggtggtca aacgtcgatg gcttttgcga tggaaggaga gacacaacag caacaaggtg
     2761 aagcaacaat tcagcgcatt gtacgtcagg caagtaagtt acgcgttgtt tttgcgacag
     2821 atgaagagat tgcagctcat gaagcccgtc tcgatctggt gcagaagaaa ggcggaagtt
     2881 gcctctggcg agcataatcc accttccagc gtttcagagc ctgccaatga gttatcaggt
     2941 cttagcccga aaatggcgcc cacaaacctt tgctgacgtc gtcggccagg aacatgtgct
     3001 gaccgcactg gcgaacggct tgtcgttagg gcgtattcat catgcttatc ttttttccgg
     3061 cacccgtggc gtcggaaaaa cctctatcgc ccgactgctg gcgaaggggc taaactgcga
     3121 aaccggcatt accgcgacgc cgtgcggcgt gtgcgataac tgtcgtgaaa tcgagcaggg
     3181 gcgctttgtc gatctgattg aaatcgacgc cgcctcgcgc accaaagttg aagatacccg
     3241 cgacctgctg gataacgtcc agtacgctcc ggcgcgtggt cgtttcaaag tttatctgat
     3301 cgacgaagtg catatgctgt cgcgccacag ctttaacgca ctgttaaaaa cccttgaaga
     3361 gccgccggag cacgttaagt ttctgctggc gacgaccgat ccacagaaat tgccggtgac
     3421 gattttgtca cgctgtctgc aatttcatct caaggcgctg gatgtcgagc aaattcgcca
     3481 tcagcttgag cacatcctca acgaagaaca tatcgctcac gagccgcggg cgctgcaatt
     3541 gctggcacgc gccgctgaag gcagcctgcg agatgcctta agtctgaccg accaggcgat
     3601 tgccagcggt gacggccagg tttcaaccca ggcggtcagt gcgatgctgg gtacgcttga
     3661 cgacgatcag gcgctgtcgc tggttgaagc gatggtcgag gccaacggcg agcgcgtaat
     3721 ggcgctgatt aatgaagccg ctgcccgtgg tatcgagtgg gaagcgttgc tggtggaaat
     3781 gctcggcctg ttgcatcgta ttgcgatggt acaactttcg cctgctgcac ttggcaacga
     3841 catggccgcc atcgagctgc ggatgcgtga actggcgcgc accataccgc cgacggatat
     3901 tcagctttac tatcagacgc tgttgattgg tcgcaaagaa ttaccgtatg cgccggaccg
     3961 tcgcatgggc gttgagatga cgctgctgcg cgcgctggca ttccatccgc gtatgccgct
     4021 gcctgagcca gaagtgccac gacagtcctt tgcacccgtc gcgccaacgg cagtaatgac
     4081 gccaacccag gtgccgccgc aaccgcaatc agcgccgcag caggcaccga ctgtaccgct
     4141 cccggaaacc accagccagg tgctggcggc gcgccagcag ttgcagcgcg tgcagggagc
     4201 aaccaaagca aaaaagagtg aaccggcagc cgctacccgc gcgcggccgg tgaataacgc
     4261 tgcgctggaa agactggctt cggtcaccga tcgcgttcag gcgcgtccgg tgccatcggc
     4321 gctggaaaaa gcgccagcca aaaaagaagc gtatcgctgg aaggcgacca ctccggtgat
     4381 gcagcaaaaa gaagtggtcg ccacgccgaa ggcgctgaaa aaagcgctgg aacatgaaaa
     4441 aacgccggaa ctggcggcga agctagcggc agaagccatt gagcgcgacc cgtgggcggc
     4501 acaggtgagc caactttcgc taccaaaact ggtcgaacag gtggcgttaa atgcctggaa
     4561 agaggagagc gacaacgcag tatgtctgca tttgcgctcc tctcagcggc atttgaacaa
     4621 ccgcggtgca cagcaaaaac tggctgaagc gttgagcatg ttaaaaggtt caacggttga
     4681 actgactatc gttgaagatg ataatcccgc ggtgcgtacg ccgctgaaat ggcgtcaggc
     4741 gatatacgaa gaaaaacttg cgcaggcgcg cgagtccatt attgcggata ataatattca
     4801 gaccctgcgt cggttcttcg atgcggagct ggatgaagaa agtatccgcc ccatttaaac
     4861 ttaattaacg gcactcctca gccaagtcaa aagcctccga ccggaggctt ttgactacat
     4921 gcccatggcg tttacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     4981 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
     5041 agcaccttgt cgccttgcgt ataatatttg cccatagtga aaacgggggc gaagaagttg
     5101 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
     5161 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     5221 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     5281 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     5341 atcaccagct caccgtcttt cattgccata cggaactccg gatgagcatt catcaggcgg
     5401 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     5461 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     5521 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     5581 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     5641 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgcca agccaaatag
     5701 gccgt
//
