LOCUS       sequence-220327-        6681 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 6681)
  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 6681)
  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: MP6
FEATURES             Location/Qualifiers
     source          1..6681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             3093..3347
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             3377..4003
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     terminator      complement(4030..4061)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4077..4736)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     rep_origin      complement(4889..5627)
                     /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      5717..5762
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(5777..6655)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac ccaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttgga
      301 cgcgtacaac tcaagtctga cataaatgac cgctatgagc actgcaatta cacgccagat
      361 cgttctcgct accgcaacca ccggtatgaa ccagattggt gcgcactatg aaggccacaa
      421 gatcattgag attggtgccg ttgaagtggt gaaccgtcgc ctgacgggca ataacttcca
      481 tgtttatctc aaacccgatc ggctggtgga tccggaagcc tttggcgtac atggtattgc
      541 cgatgaattt ttgctcgata agcccacgtt tgccgaagta gccgatgagt tcatggacta
      601 tattcgcggc gcggagttgg tgatccataa cgcagcgttc gatatcggct ttatggacta
      661 cgagttttcg ttgcttaagc gcgatattcc gaagaccaat actttctgta aggtcaccga
      721 tagccttgcg gtggcgagga aaatgtttcc cggtaagcgc aacagcctcg atgcgttatg
      781 tgctcgctac gaaatagata acagtaaacg aacgctgcac ggggcattac tcgatgccca
      841 gatccttgcg gaagtttatc tggcgatgac cggtggtcaa acgtcgatgg cttttgcgat
      901 ggaaggagag acacaacagc aacaaggtga agcaacaatt cagcgcattg tacgtcaggc
      961 aagtaagtta cgcgttgttt ttgcgacaga tgaagagatt gcagctcatg aagcccgtct
     1021 cgatctggtg cagaagaaag gcggaagttg cctctggcga gcataattta atatcagtaa
     1081 accggacata acccatgaag aaaaatcgcg cttttttgaa gtgggcaggg ggcaagtatc
     1141 ccctgcttga tgatattaaa cggcatttgc ccaagggcga atgtctggtt gagccttttg
     1201 taggtgccgg gtcggtgttt ctcaacaccg acttttctcg ttatatcctt gccgatatca
     1261 atagcgacct gatcagtctc tataacattg tgaagatgcg tactgatgag tacgtacagg
     1321 ccgcacgcga gctgtttgtt cccgaaacaa attgcgccga ggtttactat cagttccgcg
     1381 aagagttcaa caaaagccag gatccgttcc gtcgggcggt actgttttta tatttgaacc
     1441 gctacggtta caacggcctg tgtcgttaca atctgcgcgg tgagtttaac gtgccgttcg
     1501 gccgctacaa aaaaccctat ttcccggaag cagagttgta tcacttcgct gaaaaagcgc
     1561 agaatgcctt tttctattgt gagtcttacg ccgatagcat ggcgcgcgca gatgatgcat
     1621 ccgtcgtcta ttgcgatccg ccttatgcac cgctgtctgc gaccgccaac tttacggcgt
     1681 atcacacaaa cagttttacg cttgaacaac aagcgcatct ggcggagatc gccgaaggtc
     1741 tggttgagcg ccatattcca gtgctgatct ccaatcacga tacgatgtta acgcgtgagt
     1801 ggtatcagcg cgcaaaattg catgtcgtca aagttcgacg cagtataagc agcaacggcg
     1861 gcacacgtaa aaaggtggac gaactgctgg ctttgtacaa accaggagtc gtttcacccg
     1921 cgaaaaaata attcagctaa gacactgcac tggattaaga tgaaaacgat tgaagttgat
     1981 gatgaactct acagctatat tgccagccac actaagcata tcggcgagag cgcatccgac
     2041 attttacggc gtatgttgaa attttccgcc gcatcacagc ctgctgctcc ggtgacgaaa
     2101 gaggttcgcg ttgcgtcacc tgctatcgtc gaagcgaagc cggtcaaaac gattaaagac
     2161 aaggttcgcg caatgcgtga acttctgctt tcggatgaat acgcagagca aaagcgagcg
     2221 gtcaatcgct ttatgctgct gttgtctaca ctatattctc ttgacgccca ggcgtttgcc
     2281 gaagcaacgg aatcgttgca cggtcgtaca cgcgtttact ttgcggcaga tgaacaaacg
     2341 ctgctgaaaa atggtaatca gaccaagccg aaacatgtgc caggcacgcc gtattgggtg
     2401 atcaccaaca ccaacaccgg ccgtaaatgc agcatgatcg aacacatcat gcagtcgatg
     2461 caattcccgg cggaattgat tgagaaggtt tgcggaacta tctaacggct gaaattaatg
     2521 aggtcatacc caaatggata gttcgtttac gcccattgaa caaatgctaa aatttcgcgc
     2581 cagccgccac gaagattttc cttatcagga gatccttctg actcgtcttt gcatgcacat
     2641 gcaaagcaag ctgctggaga accgcaataa aatgctgaag gctcagggaa ttaacgagac
     2701 gttgtttatg gcgttgatta cgctggagtc tcaggaaaac cacagtattc agccttctga
     2761 attaagttgt gctcttggat catcccgtac caacgcgacg cgtattgccg atgaactgga
     2821 aaaacgcggt tggatcgaac gtcgtgaaag cgataacgat cgccgctgcc tgcatctgca
     2881 attaacggaa aaaggtcacg agtttttgcg cgaggtttta ccaccgcagc ataactgcct
     2941 gcatcaactc tggtccgcgc tcagcacaac agaaaaagat cagctcgagc aaatcacccg
     3001 caaattgctc tcccgtctcg accagatgga acaagacggt gtggttctcg aagcgatgag
     3061 ctaataatac aaaaattagg aggaatttca acatgacaaa tttatctgac atcattgaaa
     3121 aagaaacagg aaaacaacta gtgattcaag aatcaattct aatgttacca gaagaagtag
     3181 aggaagtaat tgggaataaa ccagaaagtg atattttagt tcatactgct tatgatgaaa
     3241 gtacagatga aaatgtaatg ctattaactt cagatgctcc agaatataaa ccttgggctt
     3301 tagtaattca agacagtaat ggagaaaata aaattaaaat gttataagtc gagattaagt
     3361 aaaccggaat ctgaagatga ccgacgcgga atacgttcgt atccacgaaa aactggacat
     3421 ctacaccttc aaaaaacagt tcttcaacaa caaaaaatct gtttctcacc gttgctacgt
     3481 tctgttcgaa ctgaaacgtc gtggtgaacg tcgtgcgtgc ttctggggtt acgcggttaa
     3541 caaaccgcag tctggtaccg aacgtggtat ccacgcggaa atcttctcta tccgtaaagt
     3601 tgaagaatac ctgcgtgaca acccgggtca gttcaccatc aactggtact cttcttggtc
     3661 tccgtgcgcg gactgcgcgg aaaaaatcct ggaatggtac aaccaggaac tgcgtggtaa
     3721 cggtcacacc ctgaaaatct gggcgtgcaa actgtactac gaaaaaaacg cgcgtaacca
     3781 gatcggtctg tggaacctgc gtgacaacgg tgttggtctg aacgttatgg tttctgaaca
     3841 ctaccagtgc tgccgtaaaa tcttcatcca gtcttctcac aaccagctga acgaaaaccg
     3901 ttggctggaa aaaaccctga aacgtgcgga aaaacgtcgt tctgaactgt ctatcatgat
     3961 ccaggttaaa atcctgcaca ccaccaaatc tccggcggtt taaacttaat taacggcact
     4021 cctcagccaa gtcaaaagcc tccggtcgga ggcttttgac tacatgccca tggcgtttac
     4081 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     4141 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct
     4201 tgcgtataat atttgcccat agtgaaaacg ggggcgaaga agttgtccat attggccacg
     4261 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     4321 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     4381 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     4441 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     4501 tctttcattg ccatacggaa ctccggatga gcattcatca ggcgggcaag aatgtgaata
     4561 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     4621 agctgaacgg tctggttata ggtacattga gtaactgact gaaatgcctc aaaatgttct
     4681 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     4741 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     4801 tcattatggt gaaagttgga acctcttacg tgccaagcca aataggccgt cactcggtcg
     4861 ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt tacccacaga
     4921 ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg
     4981 tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt ttccggtgca
     5041 tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac ccgacaggac
     5101 ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg ttccgaccct
     5161 gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc gctttctcat
     5221 agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc
     5281 aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca cttgagtcca
     5341 acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg gagttcgcag
     5401 aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt ccggctacac
     5461 tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt taagcagttc
     5521 cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg tttacagggc
     5581 aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctactgaac
     5641 cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc agcccaggag
     5701 gaagaggaca tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt
     5761 tagacttagg gaccctttat gacaacttga cggctacatc attcactttt tcttcacaac
     5821 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
     5881 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     5941 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     6001 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
     6061 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     6121 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
     6181 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     6241 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     6301 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     6361 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     6421 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     6481 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     6541 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     6601 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     6661 tcatactccc accattcaga g
//
