LOCUS       sequence-123375-        7256 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 7256)
  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 7256)
  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-D4
FEATURES             Location/Qualifiers
     source          1..7256
                     /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      6436..6467
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(6483..7142)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     7057..7076
                     /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 agcataagtc gagattaagt aaaccggaat ctgaagatgt ctgaaccacg
     2941 tttcgtacac ctgcgggtgc acagcgacta ctcgatgatc gatggcctgg ccaaaaccgc
     3001 accgttggta aaaaaggcgg cggcgttggg tatgccagca ctggcgatca ccgatttcac
     3061 caacctttgt ggtctggtga agttctacgg agcgggacat ggcgcaggga ttaagcctat
     3121 cgtcggggca gattttaacg tccagtgcga cctgctgggt gatgagttaa cccacctgac
     3181 ggtactggcg gcgaacaata ccggctatca gaatctgacg ttgctgatct caaaagcgta
     3241 tcagcgcggg tacggtgccg ccgggccgat catcgatcgc gactggctta tcgaattaaa
     3301 cgaagggttg atccttcttt ccggcggacg catgggcgac gtcggacgca gtcttttgcg
     3361 tggtaacagc gcgctggtag atgagtgtgt cgcgttttat gaagaacact tcccggatcg
     3421 ctattttctc gagctgatcc gcaccggcag gccggatgaa gaaagctatc tgcacgcggc
     3481 ggtggaactg gcggaagcgc gcggtttgcc cgtcgtggcg accaacgacg tgcgctttat
     3541 cgacagcagc gactttgacg cacacgaaat ccgcgtcgcg atccacgacg gctttaccct
     3601 cgacgatcct aaacgcccgc gtaactattc gccgcagcaa tatatgcgta gcgaagagga
     3661 gatgtgtgag ctgtttgccg acatccccga agcccttgcc aacaccgttg agatcgccaa
     3721 acgctgtaac gtaaccgtgc gtcttggtga atacttcctg ccgcagttcc cgaccgggga
     3781 catgagcacc gaagattatc tggtcaagcg tgcaaaagag ggcctggaag agcgtctggc
     3841 ctttttattc cctgatgagg aagaacgtct taagcgccgc ccggaatatg acgaacgtct
     3901 ggagactgaa cttcaggtta tcaaccagat gggcttcccg ggctacttcc tcatcgttat
     3961 ggaatttatc cagtggtcga aagataacgg cgtaccggta gggccaggcc gtggctccgg
     4021 tgcgggttca ctggtggcct acgcgctgaa aatcaccgac ctcgatccgc tggaatttga
     4081 cctgctgttc gaacgtttcc ttaacccgga acgtgtctcc atgcctgact tcgacgttga
     4141 cttctgtatg gagaaacgcg atcaggttat cgagcacgta gcggacatgt acggtcgtga
     4201 tgcggtatcg cagatcatca ccttcggtac aatggcggcg aaagcggtga tccgcgacgt
     4261 aggccgcgtg ctggggcatc cgtacggctt tgtcgatcgt atctcgaaac tgatcccgcc
     4321 cgatccgggg atgacgctgg cgaaagcgtt tgaagccgag ccgcagctgc cggaaatcta
     4381 cgaagcggat gaagaagtta aggcgctgat cgacatggcg cgcaaactgg aaggggtcac
     4441 ccgtaacgcc ggtaagcacg ccggtggggt ggttatcgcg ccgaccaaaa ttaccgattt
     4501 tgcgccgctt tactgcgatg aagagggcaa acatccggtc acccagtttg ataaaagcga
     4561 cgttgaatac gccggactgg tgaagttcga cttccttggt ttgcgtacgc tcaccatcat
     4621 caactgggcg ctggagatga tcaacaagcg gcgggcgaag aatggcgagc cgccgctgga
     4681 tatcgctgcg atcccgctgg atgataagaa aagcttcgac atgctgcaac gctcggaaac
     4741 cacggcggta ttccagcttg aatcgcgcgg catgaaggac ctgatcaagc gtctacaacc
     4801 tgactgcttc gaagatatga tcgccctagt ggcactgttc cgccccggtc cgttgcaatc
     4861 agggatggtg gataacttta tcgaccgtaa acatggtcgt gaagagatct cctatccgga
     4921 cgtacagtgg cagcatgaaa gcctgaaacc ggtactggag ccaacctacg gcattatcct
     4981 gtatcaggaa caggtcatgc agattgcgca ggtgctttct ggttataccc tcggtggcgc
     5041 ggatatgctg cgtcgtgcga tgggtaagaa aaagccggaa gagatggcta agcaacgttc
     5101 tgtatttgct gaaggtgcag aaaagaacgg aatcaacgct gaactggcga tgaaaatctt
     5161 cgacctggtg gagaaattcg ctggttacgg atttaacaaa tcgcactctg cggcctatgc
     5221 tttggtgtca tatcaaacgt tatggctgaa agcgcactat cctgcggagt ttatggcggc
     5281 ggtaatgacc gccgatatgg acaacaccga gaaggtggtg ggtctggtgg atgagtgctg
     5341 gcggatgggg ctgaaaatcc tgccaccaga tataaactcc ggtctttacc atttctatgt
     5401 caacgacgac ggcgaaatcg tgtatggtat tggcgcgatc aaaggggtcg gtgaaggtcc
     5461 gattgaggcc atcatcgaag cccgtaataa aggcggctac ttccgcgaac tgtttgatct
     5521 ctgcgcccgt accgacacca aaaagttgaa ccgtcgcgtg ctggaaaaac tgatcatgtc
     5581 cggggcgttt gaccgtcttg ggccacattg cgcagcgctg atgaactcgc tgggcgatgc
     5641 gttaaaagcg gcagatcaac acgcgaaagc ggaagctatc ggtcaggccg atatgttcgg
     5701 cgtgctggcc gaagagccgg aacaaattga acaatcctac gccagctgcc aaccgtggcc
     5761 ggagcaggtg gtattagatg gggaacgtga aacgttaggc ctgtacctga ccggacaccc
     5821 tatcaaccag tatttaaaag agattgagcg ttatgtcgga ggcgtaaggc tgaaagacat
     5881 gcacccgaca gaacgtggta aagtcatcac ggctgcgggg ctcgttgttg ccgcgcgggt
     5941 tatggtcacc aagcgcggca atcgtatcgg tatctgcacg ctggatgacc gttccgggcg
     6001 gctggaagtg atgttgttta ctgacgccct ggataaatac cagcaattgc tggaaaaaga
     6061 ccgcatactt atcgtcagcg gacaggtcag ctttgatgac ttcagcggtg ggcttaaaat
     6121 gaccgctcgc gaagtgatgg atattgacga agcccgggaa aaatatgctc gcgggcttgc
     6181 tatctcgctg acggacaggc aaattgatga ccagctttta aaccgactcc gtcagtctct
     6241 ggaaccccac cgctctggga caattccagt acatctctac tatcagaggg cggatgcacg
     6301 cgcgcggttg cgttttggcg cgacgtggcg tgtctctccg agcgatcgtt tattaaacga
     6361 tctccgtggc ctcattggtt cggagcaggt ggaactggag tttgactaaa cttaattaac
     6421 ggcactcctc agccaagtca aaagcctccg accggaggct tttgactaca tgcccatggc
     6481 gtttacgccc cgccctgcca ctcatcgcag tactgttgta attcattaag cattctgccg
     6541 acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg
     6601 tcgccttgcg tataatattt gcccatagtg aaaacggggg cgaagaagtt gtccatattg
     6661 gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata
     6721 ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc
     6781 gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac
     6841 gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc
     6901 tcaccgtctt tcattgccat acggaactcc ggatgagcat tcatcaggcg ggcaagaatg
     6961 tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta
     7021 atatccagct gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa
     7081 tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc
     7141 attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc cggtagtgat
     7201 cttatttcat tatggtgaaa gttggaacct cttacgtgcc aagccaaata ggccgt
//
