LOCUS       sequence-123400-        5364 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 5364)
  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 5364)
  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-L2
FEATURES             Location/Qualifiers
     source          1..5364
                     /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      4544..4575
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4591..5250)
                     /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 ccgctatggc gcaggcaggt tttatcttaa cccgacactg gcgggacacc ccgcaaggga
     2221 cagaagtctc cttctggctg gcgacggaca acgggccgtt gcaggttacg cttgcaccgc
     2281 aagagtccgt ggcgtttatt cccgccgatc aggttccccg cgctcagcat attttgcagg
     2341 gtgaacaagg cgttcgcctg acaccgctgg cgttaaagga ttttcaccgc cagccggtgt
     2401 atggccttta ctgtcgcgcc catcgccaat tgatgaatta cgaaaagcgc ctgcgtgaag
     2461 gtggcgttac cgtctacgag gccgatgtgc gcccgccaga acgctatctg atggagcggt
     2521 ttatcacctc accggtgtgg gtcgagggtg atatgcacaa tggcgctatc gttaatgccc
     2581 gtctgaaacc gcatcccgac tatcgtccgc cgctcaagtg ggtttctatt gcgattgaaa
     2641 ccacccgcca cggtgagctg tactgcatcg gcctggaagg ctgcgggcag cgcatcgttt
     2701 atatgctggg gccagagaat ggcgacgcct ccgcgctcga ttttgaactg gaatacgtcg
     2761 ccagccgccc gcagctactg gaaaaactca acgcctggtt tgccaactac gatcctgatg
     2821 tgatcatcgg ttggaacgtg gtgcagttcg atctgcgaat gctgcaaaaa catgccgagc
     2881 gttaccgtat tccgctgcgg ctggggcgcg ataacagtga gctggagtgg cgcgagcacg
     2941 gctttaaaaa cggcgtcttt tttgcccagg ctaaaggtcg gctaattatc gacggtatcg
     3001 aggcgctgaa atccgcgttc tggaatttct cttcattctc gctggaaacc gtcgctcagg
     3061 agttattagg cgaaggaaaa tctatcgata acccgtggga tcgaatggac gaaattgacc
     3121 gccgtttcgc cgaagataaa cctgcgctgg cgacttataa cctgaaagat tgcgagctgg
     3181 tgacgcagat tttccataaa actgaaatca tgccattttt actcgaacga gcgacggtga
     3241 acggcctgcc ggtagaccgc cacgggggat cggtggcggc gtttggtcat ctctattttc
     3301 ctcgtatgca ccgcgctggt tatgtcgcgc ctaatctcgg cgaagtgccg ccacacgcca
     3361 gccctggcgg ctacgtgatg gattcacggc cagggcttta tgattcggta ctggtgctgg
     3421 actataaaag cctgtacccg tcgatcatcc gcacctttct gattgatccc gtcgggctgg
     3481 tggaaggcat ggcgcagcct gatccagagc acagtaccga aggttttctc gatgcctggt
     3541 tctcgcgaga aaaacattgc ctgccggaga ttgtgactaa catctggcac gggcgcgatg
     3601 aagccaaacg ccagggtaac aaaccgctgt cgcaggcgct gaaaatcatc atgaatgcct
     3661 tttatggcgt gctcggcacc accgcctgcc gcttcttcga tccgcggctg gcatcgtcga
     3721 tcaccatgcg tggtcatcag atcatgcggc aaaccaaagc gttgattgaa gcacagggct
     3781 acgacgttat ctacggcgat accgactcaa cgtttgtctg gctgaaaggc gcacattcgg
     3841 aagaagaagc ggcgaaaatc ggtcgtgcac tggtgcagca cgttaacgcc tggtgggcgg
     3901 aaacgctgca aaaacaacgg ctgaccagcg cattagaact ggagtatgaa acccatttct
     3961 gccgttttct gatgccaacc attcgcggag ccgataccgg cagtaaaaag cgttatgccg
     4021 gactgattca ggagggcgac aagcagcgga tggtgtttaa agggctggaa accgtgcgca
     4081 ccgactggac gccgctggcc cagcagtttc agcaggagct atacctgcgc atcttccgca
     4141 acgagccata tcaggaatat gtacgcgaaa ccatcgacaa actgatggcg ggtgaactgg
     4201 atgcgcgact ggtttaccgt aaacgccttc gccgtccgct gagcgagtat cagcgtaatg
     4261 tgccgcctca tgtacgcgcc gctcgccttg ccgatgaaga aaaccaaaag cgtggtcgcc
     4321 ccttgcaata tcagaatcgc ggcaccatta agtacgtatg gaccaccaac ggcccggagc
     4381 cgctggacta ccaacgttca ccactggatt acgaacacta tctgacccgc cagctacaac
     4441 ccgtggcgga gggaatactc ccttttattg aggataattt tgctacactt atgaccgggc
     4501 aacttgggct attttaaact taattaacgg cactcctcag ccaagtcaaa agcctccgac
     4561 cggaggcttt tgactacatg cccatggcgt ttacgccccg ccctgccact catcgcagta
     4621 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     4681 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatagtgaa
     4741 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     4801 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     4861 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     4921 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     4981 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaactccgg
     5041 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     5101 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     5161 ttgagtaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     5221 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     5281 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     5341 tacgtgccaa gccaaatagg ccgt
//
