LOCUS       sequence-123378-        6335 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 6335)
  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 6335)
  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-F2
FEATURES             Location/Qualifiers
     source          1..6335
                     /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             3494..3505
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     terminator      5515..5546
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(5562..6221)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6136..6155
                     /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 agcataattt aatatcaggg aaccggacat aaccccatga gtgcaataga
     2941 aaatttcgac gcccatacgc ccatgatgca gcagtatctc aggctgaaag cccagcatcc
     3001 cgagatcctg ctgttttacc ggatgggtga tttttatgaa ctgttttatg acgacgcaaa
     3061 acgcgcgtcg caactgctgg atatttcact gaccaaacgc ggtgcttcgg cgggagagcc
     3121 gatcccgatg gcggggattc cctaccatgc ggtggaaaac tatctcgcca aactggtgaa
     3181 tcagggagag tccgttgcca tctgcgaaca aattggcgat ccggcgacca gcaaaggtcc
     3241 ggttgagcgc aaagttgtgc gtatcgttac gccaggcacc atcagcgatg aagccctgtt
     3301 gcaggagcgt caggacaacc tgctggcggc tatctggcag gacagcaaag gtttcggcta
     3361 cgcgacgctg gatatcagtt ccgggcgttt tcgcctgagc gaaccggctg accgcgaaac
     3421 gatggcggca gaactgcaac gcactaatcc tgcggaactg ctgtatgcag aagattttgc
     3481 tgaaatgtcg ttaattgaag gccgtcgcgg cctgcgccgt cgcccgctgt gggagtttga
     3541 aatcgacacc gcgcgccagc agttgaatct gcaatttggg acccgcgatc tggtcggttt
     3601 tggcgtcgag aacgcgccgc gcggactttg tgctgccggt tgtctgttgc agtatgcgaa
     3661 agatacccaa cgtacgactc tgccgcatat tcgttccatc accatggaac gtgagcagga
     3721 cagcatcatt atggatgccg cgacgcgtcg taatctggaa atcacccaga acctggcggg
     3781 tggtgcggaa aatacgctgg cttctgtgct cgactgcacc gtcacgccga tgggcagccg
     3841 tatgctgaaa cgctggctgc atatgccagt gcgcgatacc cgcgtgttgc ttgagcgcca
     3901 gcaaactatt ggcgcattgc aggatttcac cgccgggcta cagccggtac tgcgtcaggt
     3961 cggcgacctg gaacgtattc tggcacgtct ggctttacga actgctcgcc cacgcgatct
     4021 ggcccgtatg cgccacgctt tccagcaact gccggagctg cgtgcgcagt tagaaactgt
     4081 cgatagtgca ccggtacagg cgctacgtga gaagatgggc gagtttgccg agctgcgcga
     4141 tctgctggag cgagcaatca tcgacacacc gccggtgctg gtacgcgacg gtggtgttat
     4201 cgcatcgggc tataacgaag agctggatga gtggcgcgcg ctggctgacg gcgcgaccga
     4261 ttatctggag cgtctggaag tccgcgagcg tgaacgtacc ggcctggaca cgctgaaagt
     4321 tggctttaat gcggtgcacg gctactacat tcaaatcagc cgtgggcaaa gccatctggc
     4381 acccatcaac tacatgcgtc gccagacgct gaaaaacgcc gagcgctaca tcattccaga
     4441 gctaaaagag tacgaagata aagttctcac ctcaaaaggc aaagcactgg cactggaaaa
     4501 acagctttat gaagagctgt tcgacctgct gttgccgcat ctggaagcgt tgcaacagag
     4561 cgcgagcgcg ctggcggaac tcgacgtgct ggttaacctg gcggaacggg cctataccct
     4621 gaactacacc tgcccgacct tcattgataa accgggcatt cgcattaccg aaggtcgcca
     4681 tccggtagtt gaacaagtac tgaatgagcc atttatcgcc aacccgctga atctgtcgcc
     4741 gcagcgccgc atgttgatca tcaccggtcc gaacatgggc ggtaaaagta cctatatgcg
     4801 ccagaccgca ctgattgcgc tgatggccta catcggcagc tatgtaccgg cacaaaaagt
     4861 cgagattgga cctatcgatc gcatctttac ccgcgtaggc gcggcagatg acctggcgtc
     4921 cgggcgctca acctttatgg tggagatgac tgaaaccgcc aatattttac ataacgccac
     4981 cgaatacagt ctggtgttaa tggtggagat cgggcgtgga acgtccacct acgatggtct
     5041 gtcgctggcg tgggcgtgcg cggaaaatct ggcgaataag attaaggcat tgacgttatt
     5101 tgctacccac tatttcgagc tgacccagtt accggagaaa atggaaggcg tcgctaacgt
     5161 gcatctcgat gcactggagc acggcgacac cattgccttt atgcacagcg tgcaggatgg
     5221 cgcggcgagc aaaagctacg gcctggcggt tgcagctctg gcaggcgtgc caaaagaggt
     5281 tattaagcgc gcacggcaaa agctgcgtga gctggaaagc atttcgccga acgccgccgc
     5341 tacgcaagtg gatggtacgc aaatgtcttt gctgtcagta ccagaagaaa cttcgcctgc
     5401 ggtcgaagct ctggaaaatc ttgatccgga ttcactcacc ccgcgtcagg cgctggagtg
     5461 gatttatcgc ttgaagagcc tggtgtaaac ttaattaacg gcactcctca gccaagtcaa
     5521 aagcctccga ccggaggctt ttgactacat gcccatggcg tttacgcccc gccctgccac
     5581 tcatcgcagt actgttgtaa ttcattaagc attctgccga catggaagcc atcacaaacg
     5641 gcatgatgaa cctgaatcgc cagcggcatc agcaccttgt cgccttgcgt ataatatttg
     5701 cccatagtga aaacgggggc gaagaagttg tccatattgg ccacgtttaa atcaaaactg
     5761 gtgaaactca cccagggatt ggctgagacg aaaaacatat tctcaataaa ccctttaggg
     5821 aaataggcca ggttttcacc gtaacacgcc acatcttgcg aatatatgtg tagaaactgc
     5881 cggaaatcgt cgtggtattc actccagagc gatgaaaacg tttcagtttg ctcatggaaa
     5941 acggtgtaac aagggtgaac actatcccat atcaccagct caccgtcttt cattgccata
     6001 cggaactccg gatgagcatt catcaggcgg gcaagaatgt gaataaaggc cggataaaac
     6061 ttgtgcttat ttttctttac ggtctttaaa aaggccgtaa tatccagctg aacggtctgg
     6121 ttataggtac attgagcaac tgactgaaat gcctcaaaat gttctttacg atgccattgg
     6181 gatatatcaa cggtggtata tccagtgatt tttttctcca ttttagcttc cttagctcct
     6241 gaaaatctcg ataactcaaa aaatacgccc ggtagtgatc ttatttcatt atggtgaaag
     6301 ttggaacctc ttacgtgcca agccaaatag gccgt
//
