LOCUS       sequence-123370-        6495 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 6495)
  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 6495)
  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-B4
FEATURES             Location/Qualifiers
     source          1..6495
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      3530..3561
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(3577..4236)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4151..4170
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      complement(4389..5127)
                     /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      5217..5262
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5277..6155)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        6182..6466
                     /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     6389..6408
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 gtctgacata aatgaccgct atgtctgaac cacgtttcgt acacctgcgg gtgcacagcg
       61 actactcgat gatcgatggc ctggccaaaa ccgcaccgtt ggtaaaaaag gcggcggcgt
      121 tgggtatgcc agcactggcg atcaccgatt tcaccaacct ttgtggtctg gtgaagttct
      181 acggagcggg acatggcgca gggattaagc ctatcgtcgg ggcagatttt aacgtccagt
      241 gcgacctgct gggtgatgag ttaacccacc tgacggtact ggcggcgaac aataccggct
      301 atcagaatct gacgttgctg atctcaaaag cgtatcagcg cgggtacggt gccgccgggc
      361 cgatcatcga tcgcgactgg cttatcgaat taaacgaagg gttgatcctt ctttccggcg
      421 gacgcatggg cgacgtcgga cgcagtcttt tgcgtggtaa cagcgcgctg gtagatgagt
      481 gtgtcgcgtt ttatgaagaa cacttcccgg atcgctattt tctcgagctg atccgcaccg
      541 gcaggccgga tgaagaaagc tatctgcacg cggcggtgga actggcggaa gcgcgcggtt
      601 tgcccgtcgt ggcgaccaac gacgtgcgct ttatcgacag cagcgacttt gacgcacacg
      661 aaatccgcgt cgcgatccac gacggcttta ccctcgacga tcctaaacgc ccgcgtaact
      721 attcgccgca gcaatatatg cgtagcgaag aggagatgtg tgagctgttt gccgacatcc
      781 ccgaagccct tgccaacacc gttgagatcg ccaaacgctg taacgtaacc gtgcgtcttg
      841 gtgaatactt cctgccgcag ttcccgaccg gggacatgag caccgaagat tatctggtca
      901 agcgtgcaaa agagggcctg gaagagcgtc tggccttttt attccctgat gaggaagaac
      961 gtcttaagcg ccgcccggaa tatgacgaac gtctggagac tgaacttcag gttatcaacc
     1021 agatgggctt cccgggctac ttcctcatcg ttatggaatt tatccagtgg tcgaaagata
     1081 acggcgtacc ggtagggcca ggccgtggct ccggtgcggg ttcactggtg gcctacgcgc
     1141 tgaaaatcac cgacctcgat ccgctggaat ttgacctgct gttcgaacgt ttccttaacc
     1201 cggaacgtgt ctccatgcct gacttcgacg ttgacttctg tatggagaaa cgcgatcagg
     1261 ttatcgagca cgtagcggac atgtacggtc gtgatgcggt atcgcagatc atcaccttcg
     1321 gtacaatggc ggcgaaagcg gtgatccgcg acgtaggccg cgtgctgggg catccgtacg
     1381 gctttgtcga tcgtatctcg aaactgatcc cgcccgatcc ggggatgacg ctggcgaaag
     1441 cgtttgaagc cgagccgcag ctgccggaaa tctacgaagc ggatgaagaa gttaaggcgc
     1501 tgatcgacat ggcgcgcaaa ctggaagggg tcacccgtaa cgccggtaag cacgccggtg
     1561 gggtggttat cgcgccgacc aaaattaccg attttgcgcc gctttactgc gatgaagagg
     1621 gcaaacatcc ggtcacccag tttgataaaa gcgacgttga atacgccgga ctggtgaagt
     1681 tcgacttcct tggtttgcgt acgctcacca tcatcaactg ggcgctggag atgatcaaca
     1741 agcggcgggc gaagaatggc gagccgccgc tggatatcgc tgcgatcccg ctggatgata
     1801 agaaaagctt cgacatgctg caacgctcgg aaaccacggc ggtattccag cttgaatcgc
     1861 gcggcatgaa ggacctgatc aagcgtctac aacctgactg cttcgaagat atgatcgccc
     1921 tagtggcact gttccgcccc ggtccgttgc aatcagggat ggtggataac tttatcgacc
     1981 gtaaacatgg tcgtgaagag atctcctatc cggacgtaca gtggcagcat gaaagcctga
     2041 aaccggtact ggagccaacc tacggcatta tcctgtatca ggaacaggtc atgcagattg
     2101 cgcaggtgct ttctggttat accctcggtg gcgcggatat gctgcgtcgt gcgatgggta
     2161 agaaaaagcc ggaagagatg gctaagcaac gttctgtatt tgctgaaggt gcagaaaaga
     2221 acggaatcaa cgctgaactg gcgatgaaaa tcttcgacct ggtggagaaa ttcgctggtt
     2281 acggatttaa caaatcgcac tctgcggcct atgctttggt gtcatatcaa acgttatggc
     2341 tgaaagcgca ctatcctgcg gagtttatgg cggcggtaat gaccgccgat atggacaaca
     2401 ccgagaaggt ggtgggtctg gtggatgagt gctggcggat ggggctgaaa atcctgccac
     2461 cagatataaa ctccggtctt taccatttcc acgtcaacga cgacggcgaa atcgtgtatg
     2521 gtattggcgc gatcaaaggg gtcggtgaag gtccgattga ggccatcatc gaagcccgta
     2581 ataaaggcgg ctacttccgc gaactgtttg atctctgcgc ccgtaccgac accaaaaagt
     2641 tgaaccgtcg cgtgctggaa aaactgatca tgccgggggc gtttgaccgt cttgggccac
     2701 atcgcgcagc gctgatgaac tcgctgggcg atgcgttaaa agcggcagat caacacgcga
     2761 aagcggaagc tatcggtcag gccgatatgt tcggcgtgct ggccgaagag ccggaacaaa
     2821 ttgaacaatc ctacgccagc tgccaaccgt ggccggagca ggtggtatta gatggggaac
     2881 gtgaaacgtt aggcctgtac ctgaccggac accctatcaa ccagtattta aaagagattg
     2941 agcgttatgt cggaggcgta aggctgaaag acatgcaccc gacagaacgt ggtaaagtca
     3001 tcacggctgc ggggctcgtt gttgccgcgc gggttatggt caccaagcgc ggcaatcgta
     3061 tcggtatctg cacgctggat gaccgttccg ggcggctgga agtgatgttg tttactgacg
     3121 ccctggataa ataccagcaa ttgctggaaa aagaccgcat acttatcgtc agcggacagg
     3181 tcagctttga tgacttcagc ggtgggctta aaatgaccgc tcgcgaagtg atggatattg
     3241 acgaagcccg ggaaaaatat gctcgcgggc ttgctatctc gctgacggac aggcaaattg
     3301 atgaccagct tttaaaccga ctccgtcagt ctctggaacc ccaccgctct gggacaattc
     3361 cagtacatct ctactatcag agggcggatg cacgcgcgcg gttgcgtttt ggcgcgacgt
     3421 ggcgtgtctc tccgagcgat cgtttattaa acgatctccg tggcctcatt ggttcggagc
     3481 aggtggaact ggagtttgac taaacttaat taacggcact cctcagccaa gtcaaaagcc
     3541 tccgaccgga ggcttttgac tacatgccca tggcgtttac gccccgccct gccactcatc
     3601 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     3661 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     3721 agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     3781 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     3841 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     3901 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     3961 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa
     4021 ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     4081 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     4141 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     4201 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     4261 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     4321 acctcttacg tgccaagcca aataggccgt cactcggtcg ctacgctccg ggcgtgagac
     4381 tgcggcgggc gctgcggaca catacaaagt tacccacaga ttccgtggat aagcagggga
     4441 ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg tttttccata ggctccgccc
     4501 tcctgccaga gttcacataa acagacgctt ttccggtgca tctgtgggag ccgtgaggct
     4561 caaccatgaa tctgacagta cgggcgaaac ccgacaggac ttaaagatcc ccaccgtttc
     4621 cggcgggtcg ctccctcttg cgctctcctg ttccgaccct gccgtttacc ggatacctgt
     4681 tccgcctttc tcccttacgg gaagtgtggc gctttctcat agctcacaca ctggtatctc
     4741 ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc aagaactccc cgttcagccc
     4801 gactgctgcg ccttatccgg taactgttca cttgagtcca acccggaaaa gcacggtaaa
     4861 acgccactgg cagcagccat tggtaactgg gagttcgcag aggatttgtt tagctaaaca
     4921 cgcggttgct cttgaagtgt gcgccaaagt ccggctacac tggaaggaca gatttggttg
     4981 ctgtgctctg cgaaagccag ttaccacggt taagcagttc cccaactgac ttaaccttcg
     5041 atcaaaccac ctccccaggt ggttttttcg tttacagggc aaaagattac gcgcagaaaa
     5101 aaaggatctc aagaagatcc tttgatcttt tctactgaac cgctctagat ttcagtgcaa
     5161 tttatctctt caaatgtagc acctgaagtc agcccaggag gaagaggaca tccggtcaaa
     5221 taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tagacttagg gaccctttat
     5281 gacaacttga cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg
     5341 ctggccccgg tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca
     5401 ttgcgaccga cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg
     5461 atacgttggt cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt
     5521 gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg
     5581 tctgccaggt gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga
     5641 tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc
     5701 gccagcagct ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg
     5761 tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt
     5821 gacggccagt taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga
     5881 taccattcgc gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc
     5941 aaaatatcac ccggtcggca aacaaattct cgtccctgat ttttcaccac cccctgaccg
     6001 cgaatggtga gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg
     6061 agataaccgt tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat
     6121 cccggcagca ggggatcatt ttgcgcttca gccatacttt tcatactccc accattcaga
     6181 gaagaaacca attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc
     6241 ttctcgctaa cccaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc
     6301 aaagccatga caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt
     6361 gattatttgc acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg
     6421 gatcctacct gacgcttttt atcgcaactc tctactgttt ctccataccc gtttttttgg
     6481 acgcgtacaa ctcaa
//
