LOCUS       sequence-123407-        6651 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 6651)
  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 6651)
  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-P8
FEATURES             Location/Qualifiers
     source          1..6651
                     /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"
     CDS             4924..5178
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5208..5804
                     /codon_start=1
                     /gene="AICDA"
                     /product="human activation-induced cytidine deaminase 
                     (Dickerson et al., 2003)"
                     /label=AID
                     /translation="MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLD
                     FGYLRNKNGCHVELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNL
                     SLRIFTARLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWE
                     GLHENSVRLSRQLRRILLPLYEVDDLRDAFRTLGL"
     terminator      5831..5862
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(5878..6537)
                     /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 atctaacggc tgaaattaat gaggtcatac ccaaatggat agttcgttta
     4381 cgcccattga acaaatgcta aaatttcgcg ccagccgcca cgaagatttt ccttatcagg
     4441 agatccttct gactcgtctt tgcatgcaca tgcaaagcaa gctgctggag aaccgcaata
     4501 aaatgctgaa ggctcaggga attaacgaga cgttgtttat ggcgttgatt acgctggagt
     4561 ctcaggaaaa ccacagtatt cagccttctg aattaagttg tgctcttgga tcatcccgta
     4621 ccaacgcgac gcgtattgcc gatgaactgg aaaaacgcgg ttggatcgaa cgtcgtgaaa
     4681 gcgataacga tcgccgctgc ctgcatctgc aattaacgga aaaaggtcac gagtttttgc
     4741 gcgaggtttt accaccgcag cataactgcc tgcatcaact ctggtccgcg ctcagcacaa
     4801 cagaaaaaga tcagctcgag caaatcaccc gcaaattgct ctcccgtctc gaccagatgg
     4861 aacaagacgg tgtggttctc gaagcgatga gctaataata caaaaattag gaggaatttc
     4921 aacatgacaa atttatctga catcattgaa aaagaaacag gaaaacaact agtgattcaa
     4981 gaatcaattc taatgttacc agaagaagta gaggaagtaa ttgggaataa accagaaagt
     5041 gatattttag ttcatactgc ttatgatgaa agtacagatg aaaatgtaat gctattaact
     5101 tcagatgctc cagaatataa accttgggct ttagtaattc aagacagtaa tggagaaaat
     5161 aaaattaaaa tgttataagt cgagattaag taaaccggaa tctgaagatg gactctctgc
     5221 tgatgaaccg tcgtaaattc ctgtaccagt tcaaaaacgt tcgttgggcg aaaggtcgtc
     5281 gtgaaaccta cctgtgctac gttgttaaac gtcgtgactc tgcgacctct ttctctctgg
     5341 acttcggtta cctgcgtaac aaaaacggtt gccacgttga actgctgttc ctgcgttaca
     5401 tctctgactg ggacctggac ccgggtcgtt gctaccgtgt tacctggttc acctcttggt
     5461 ctccgtgcta cgactgcgcg cgtcacgttg cggacttcct gcgtggtaac ccgaacctgt
     5521 ctctgcgtat cttcaccgcg cgtctgtact tctgcgaaga ccgtaaagcg gaaccggaag
     5581 gtctgcgtcg tctgcaccgt gcgggtgttc agatcgcgat catgaccttc aaagactact
     5641 tctactgctg gaacaccttc gttgaaaacc acgaacgtac cttcaaagcg tgggaaggtc
     5701 tgcacgaaaa ctctgttcgt ctgtctcgtc agctgcgtcg tatcctgctg ccgctgtacg
     5761 aagttgacga cctgcgtgac gcgttccgta ccctgggtct gtaaacttaa ttaacggcac
     5821 tcctcagcca agtcaaaagc ctccgaccgg aggcttttga ctacatgccc atggcgttta
     5881 cgccccgccc tgccactcat cgcagtactg ttgtaattca ttaagcattc tgccgacatg
     5941 gaagccatca caaacggcat gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc
     6001 ttgcgtataa tatttgccca tagtgaaaac gggggcgaag aagttgtcca tattggccac
     6061 gtttaaatca aaactggtga aactcaccca gggattggct gagacgaaaa acatattctc
     6121 aataaaccct ttagggaaat aggccaggtt ttcaccgtaa cacgccacat cttgcgaata
     6181 tatgtgtaga aactgccgga aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc
     6241 agtttgctca tggaaaacgg tgtaacaagg gtgaacacta tcccatatca ccagctcacc
     6301 gtctttcatt gccatacgga actccggatg agcattcatc aggcgggcaa gaatgtgaat
     6361 aaaggccgga taaaacttgt gcttattttt ctttacggtc tttaaaaagg ccgtaatatc
     6421 cagctgaacg gtctggttat aggtacattg agtaactgac tgaaatgcct caaaatgttc
     6481 tttacgatgc cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt
     6541 agcttcctta gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat
     6601 ttcattatgg tgaaagttgg aacctcttac gtgccaagcc aaataggccg t
//
