LOCUS       sequence-123421-        8861 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 8861)
  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 8861)
  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-Q11
FEATURES             Location/Qualifiers
     source          1..8861
                     /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      8041..8072
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(8088..8747)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
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 agcataattt aatatcagta aaccggacat aacccatgaa gaaaaatcgc
     2941 gcttttttga agtgggcagg gggcaagtat cccctgcttg atgatattaa acggcatttg
     3001 cccaagggcg aatgtctggt tgagcctttt gtaggtgccg ggtcggtgtt tctcaacacc
     3061 gacttttctc gttatatcct tgccgatatc aatagcgacc tgatcagtct ctataacatt
     3121 gtgaagatgc gtactgatga gtacgtacag gccgcacgcg agctgtttgt tcccgaaaca
     3181 aattgcgccg aggtttacta tcagttccgc gaagagttca acaaaagcca ggatccgttc
     3241 cgtcgggcgg tactgttttt atatttgaac cgctacggtt acaacggcct gtgtcgttac
     3301 aatctgcgcg gtgagtttaa cgtgccgttc ggccgctaca aaaaacccta tttcccggaa
     3361 gcagagttgt atcacttcgc tgaaaaagcg cagaatgcct ttttctattg tgagtcttac
     3421 gccgatagca tggcgcgcgc agatgatgca tccgtcgtct attgcgatcc gccttatgca
     3481 ccgctgtctg cgaccgccaa ctttacggcg tatcacacaa acagttttac gcttgaacaa
     3541 caagcgcatc tggcggagat cgccgaaggt ctggttgagc gccatattcc agtgctgatc
     3601 tccaatcacg atacgatgtt aacgcgtgag tggtatcagc gcgcaaaatt gcatgtcgtc
     3661 aaagttcgac gcagtataag cagcaacggc ggcacacgta aaaaggtgga cgaactgctg
     3721 gctttgtaca aaccaggagt cgtttcaccc gcgaaaaaat aattcagcta agacactgca
     3781 ctggattaag atgaaaacga ttgaagttga tgatgaactc tacagctata ttgccagcca
     3841 cactaagcat atcggcgaga gcgcatccga cattttacgg cgtatgttga aattttccgc
     3901 cgcatcacag cctgctgctc cggtgacgaa agaggttcgc gttgcgtcac ctgctatcgt
     3961 cgaagcgaag ccggtcaaaa cgattaaaga caaggttcgc gcaatgcgtg aacttctgct
     4021 ttcggatgaa tacgcagagc aaaagcgagc ggtcaatcgc tttatgctgc tgttgtctac
     4081 actatattct cttgacgccc aggcgtttgc cgaagcaacg gaatcgttgc acggtcgtac
     4141 acgcgtttac tttgcggcag atgaacaaac gctgctgaaa aatggtaatc agaccaagcc
     4201 gaaacatgtg ccaggcacgc cgtattgggt gatcaccaac accaacaccg gccgtaaatg
     4261 cagcatgatc gaacacatca tgcagtcgat gcaattcccg gcggaattga ttgagaaggt
     4321 ttgcggaact atctaaaaga attccaaaaa caggtacgac atacatgaat cagaatctgc
     4381 tggtgacaaa gcgcgacggt agcacagagc gcatcaatct cgacaaaatc catcgcgttc
     4441 tggattgggc ggcagaagga ctgcataacg tttcgatttc ccaggtcgag ctgcgctccc
     4501 acattcagtt ttatgacggt atcaagacct ctgacatcca cgaaaccatt atcaaggctg
     4561 ccgcagacct gatctcccgt gatgcgccgg attatcagta tctcgccgcg cgcctggcga
     4621 tcttccacct gcgtaaaaaa gcctacggcc agtttgagcc gcctgcgctg tacgaccacg
     4681 tggtgaaaat ggtcgagatg ggcaaatacg ataatcatct gctggaagac tacacggaag
     4741 aagagttcaa gcagatggac acctttatcg atcacgaccg tgatatgacc ttctcttatg
     4801 ctgccgttaa gcagctggaa ggcaaatatc tggtacagaa ccgcgtgacc ggcgaaatct
     4861 atgagagcgc ccagttcctt tatattctag ttgccgcgtg cttgttctcg aactacccgc
     4921 gtgaaacgcg cctgcaatat gtgaagcgtt tttacgacgc ggtttccaca tttaaaattt
     4981 cgctgccgac gccaatcatg tccggcgtgc gtaccccgac tcgtcagttc agctcctgcg
     5041 tactgatcga gtgcggtgac agcctggatt ccatcaacgc cacctccagc gcgattgtta
     5101 aatacgtttc ccagcgtgcc gggatcggca tcaacgccgg gcgtattcgt gcgctgggta
     5161 gcccgattcg cggtggtgaa gcgttccata ccggctgcat tccgttctac aaacatttcc
     5221 agacagcggt gaaatcctgc tctcagggcg gtgtgcgcgg cggtgtggca acgctgttct
     5281 acccgatgtg gcatctggaa gtggaaagcc tgctggtgtt gaaaaacaac cgtggtgtgg
     5341 aaggcaaccg cgtgcgtcat atggactacg gggtacaaat caacaaactg atgtataccc
     5401 gtctgctgaa aggtgaagat atcaccctgt tcagcccgtc cgacgtaccg gggctgtacg
     5461 acgcgttctt cgccgatcag gaagagtttg aacgtctgta taccaaatat gagaaagacg
     5521 acagcatccg caagcagcgt gtgaaagccg ttgagctgtt ctcgctgatg atgcaggaac
     5581 gtgcgtctac cggtcgtatc tatattcaga acgttgacca ctgcaatacc catagcccgt
     5641 ttgatccggc catcgcgcca gtgcgtcagt ctaacctgtg cctggagata gccctgccga
     5701 ccaaaccgct gaacgacgtc aacgacgaga acggtgaaat cgcgctgtgt acgctgtctg
     5761 ctttcaacct gggcgcaatt aataacctgg atgaactgga agagctggca attctggcgg
     5821 ttcgtgcact tgacgcgctg ctggattatc aggattaccc gatcccggcc gccaaacgtg
     5881 gagcgatggg tcgtcgtacg ctgggtattg gtgtgatcaa cttcgcttac tacctggcga
     5941 agcacggtaa acgctactcc gacggcagcg ccaacaacct gacgcataaa accttcgaag
     6001 ccattcagta ttacctgctg aaagcctcta atgagctggc gaaagagcaa ggcgcgtgcc
     6061 cgtggtttaa cgaaaccact tacgcgaaag ggatcctgcc gatcgatacc tataagaaag
     6121 atctggatac catcgctaat gagccgctgc attacgactg ggaagctctg cgtgagtcaa
     6181 tcaaaacgca cggtctgcgt aactccacgc tttctgctct gatgccgtcc gagacttctt
     6241 cgcagatctc taacgccact aacggtattg aaccgccgcg cggttacgtc agcatcaaag
     6301 cgtcgaaaga cggtattttg cgccaggtgg tgccggacta cgagcacctg cacgacgcct
     6361 atgagctgct gtgggaaatg ccgggtaacg atggttatct gcaactggtg ggtatcatgc
     6421 agaaatttat cgatcagtcg atctctgcca acaccaacta cgatccgtca cgcttcccgt
     6481 caggaaaagt gccgatgcag cagttgctga aagacctgct caccgcctac aaattcgggg
     6541 tcaaaacact gtattatcag aacacccgtg acggcgctga agacgcacaa gacgatctgg
     6601 tgccgtcaat ccaggacgat ggctgcgaaa gcggcgcatg taagatctga tattgagatg
     6661 ccggatgcgg cgtaaacgcc ttatccggcc tacggctcgg tttgtaggcc tgataagacg
     6721 cgccagcgtc gcatcaggct ccgggtgccg gatgcagcgt gaacgcctta tccggcctac
     6781 ggctcggatt tgtaggcctg ataagacgcg ccagcgtcgc atcaggcaca ggatgcggcg
     6841 taaaatgcct tatccggcat taaactccca acaggacaca ctcatggcat ataccacctt
     6901 ttcacagacg aaaaatgatc agctcaaaga accgatgttc tttggtcagc cggtcaacgt
     6961 ggctcgctac gatcagcaaa aatatgacat cttcgaaaag ctgatcgaaa agcagctctc
     7021 tttcttctgg cgtccggaag aagttgacgt ctcccgcgac cgtatagatt accaggcgct
     7081 gccggagcac gaaaaacaca tctttatcag caacctgaaa tatcagacgc tgctggattc
     7141 cattcagggt cgtagcccga acgtggcgct attgccgctt atttctattc cggaactgga
     7201 aacctgggtc gaaacctggg cgttctcaga aacgattcat tcccgttcct atactcatat
     7261 cattcgtaat atcgttaacg atccgtctgt tgtgtttgac gatatcgtca ccaacgagca
     7321 gatccagaaa cgtgcggaag ggatctccag ctattacgat gagctgatcg aaatgaccag
     7381 ctactggcat ctgctgggcg aaggtaccca caccgttaac ggtaaaactg tgaccgttag
     7441 cctgcgcgag ctgaagaaaa aactgtatct ctgcctgatg agcgttaacg cgctggaagc
     7501 gattcgtttc tacgtcagct ttgcttgttc cttcgcattt gcagaacgcg aattgatgga
     7561 aggcaacgcc aaaattattc gcctgattgc ccgcgacgaa gccctgcacc tgaccggcac
     7621 ccagcatatg ctgaatctgc tgcgcagcgg cgcggacgat cctgagatgg cggaaattgc
     7681 cgaagagtgt aagcaggagt gctatgacct gtttgttcag gcagctcaac aggagaaaga
     7741 ctgggcggat tatctgttcc gcgacggttc gatgattggt ctgaataaag acattctctg
     7801 ccagtacgtt gaatacatca ccaatatccg tatgcaggca gtcggtttgg atctgccgtt
     7861 ccagacgcgc tccaacccga tcccgtggat caacacttgg ctggtgtctg ataacgtgca
     7921 ggttgctccg caggaagtgg aagtcagttc ttatctggtc gggcagattg actcggaagt
     7981 ggacaccgac gatttgagta acttccagct ctgaacttaa ttaacggcac tcctcagcca
     8041 agtcaaaagc ctccgaccgg aggcttttga ctacatgccc atggcgttta cgccccgccc
     8101 tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca
     8161 caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa
     8221 tatttgccca tagtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca
     8281 aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct
     8341 ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga
     8401 aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca
     8461 tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt
     8521 gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga
     8581 taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg
     8641 gtctggttat aggtacattg agtaactgac tgaaatgcct caaaatgttc tttacgatgc
     8701 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
     8761 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     8821 tgaaagttgg aacctcttac gtgccaagcc aaataggccg t
//
