LOCUS       sequence-123371-        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-B5
FEATURES             Location/Qualifiers
     source          1..6495
                     /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      5675..5706
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(5722..6381)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     6296..6315
                     /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 ccgctatgtc tgaaccacgt ttcgtacacc tgcgggtgca cagcgactac tcgatgatcg
     2221 atggcctggc caaaaccgca ccgttggtaa aaaaggcggc ggcgttgggt atgccagcac
     2281 tggcgatcac cgatttcacc aacctttgtg gtctggtgaa gttctacgga gcgggacatg
     2341 gcgcagggat taagcctatc gtcggggcag attttaacgt ccagtgcgac ctgctgggtg
     2401 atgagttaac ccacctgacg gtactggcgg cgaacaatac cggctatcag aatctgacgt
     2461 tgctgatctc aaaagcgtat cagcgcgggt acggtgccgc cgggccgatc atcgatcgcg
     2521 actggcttat cgaattaaac gaagggttga tccttctttc cggcggacgc atgggcgacg
     2581 tcggacgcag tcttttgcgt ggtaacagcg cgctggtaga tgagtgtgtc gcgttttatg
     2641 aagaacactt cccggatcgc tattttctcg agctgatccg caccggcagg ccggatgaag
     2701 aaagctatct gcacgcggcg gtggaactgg cggaagcgcg cggtttgccc gtcgtggcga
     2761 ccaacgacgt gcgctttatc gacagcagcg actttgacgc acacgaaatc cgcgtcgcga
     2821 tccacgacgg ctttaccctc gacgatccta aacgcccgcg taactattcg ccgcagcaat
     2881 atatgcgtag cgaagaggag atgtgtgagc tgtttgccga catccccgaa gcccttgcca
     2941 acaccgttga gatcgccaaa cgctgtaacg taaccgtgcg tcttggtgaa tacttcctgc
     3001 cgcagttccc gaccggggac atgagcaccg aagattatct ggtcaagcgt gcaaaagagg
     3061 gcctggaaga gcgtctggcc tttttattcc ctgatgagga agaacgtctt aagcgccgcc
     3121 cggaatatga cgaacgtctg gagactgaac ttcaggttat caaccagatg ggcttcccgg
     3181 gctacttcct catcgttatg gaatttatcc agtggtcgaa agataacggc gtaccggtag
     3241 ggccaggccg tggctccggt gcgggttcac tggtggccta cgcgctgaaa atcaccgacc
     3301 tcgatccgct ggaatttgac ctgctgttcg aacgtttcct taacccggaa cgtgtctcca
     3361 tgcctgactt cgacgttgac ttctgtatgg agaaacgcga tcaggttatc gagcacgtag
     3421 cggacatgta cggtcgtgat gcggtatcgc agatcatcac cttcggtaca atggcggcga
     3481 aagcggtgat ccgcgacgta ggccgcgtgc tggggcatcc gtacggcttt gtcgatcgta
     3541 tctcgaaact gatcccgccc gatccgggga tgacgctggc gaaagcgttt gaagccgagc
     3601 cgcagctgcc ggaaatctac gaagcggatg aagaagttaa ggcgctgatc gacatggcgc
     3661 gcaaactgga aggggtcacc cgtaacgccg gtaagcacgc cggtggggtg gttatcgcgc
     3721 cgaccaaaat taccgatttt gcgccgcttt actgcgatga agagggcaaa catccggtca
     3781 cccagtttga taaaagcgac gttgaatacg ccggactggt gaagttcgac ttccttggtt
     3841 tgcgtacgct caccatcatc aactgggcgc tggagatgat caacaagcgg cgggcgaaga
     3901 atggcgagcc gccgctggat atcgctgcga tcccgctgga tgataagaaa agcttcgaca
     3961 tgctgcaacg ctcggaaacc acggcggtat tccagcttga atcgcgcggc atgaaggacc
     4021 tgatcaagcg tctacaacct gactgcttcg aagatatgat cgccctagtg gcactgttcc
     4081 gccccggtcc gttgcaatca gggatggtgg ataactttat cgaccgtaaa catggtcgtg
     4141 aagagatctc ctatccggac gtacagtggc agcatgaaag cctgaaaccg gtactggagc
     4201 caacctacgg cattatcctg tatcaggaac aggtcatgca gattgcgcag gtgctttctg
     4261 gttataccct cggtggcgcg gatatgctgc gtcgtgcgat gggtaagaaa aagccggaag
     4321 agatggctaa gcaacgttct gtatttgctg aaggtgcaga aaagaacgga atcaacgctg
     4381 aactggcgat gaaaatcttc gacctggtgg agaaattcgc tggttacgga tttaacaaat
     4441 cgcactctgc ggcctatgct ttggtgtcat atcaaacgtt atggctgaaa gcgcactatc
     4501 ctgcggagtt tatggcggcg gtaatgaccg ccgatatgga caacaccgag aaggtggtgg
     4561 gtctggtgga tgagtgctgg cggatggggc tgaaaatcct gccaccagat ataaactccg
     4621 gtctttacca tttctatgtc aacgacgacg gcgaaatcgt gtatggtatt ggcgcgatca
     4681 aaggggtcgg tgaaggtccg attgaggcca tcatcgaagc ccgtaataaa ggcggctact
     4741 tccgcgaact gtttgatctc tgcgcccgta ccgacaccaa aaagttgaac cgtcgcgtgc
     4801 tggaaaaact gatcatgtcc ggggcgtttg accgtcttgg gccacattgc gcagcgctga
     4861 tgaactcgct gggcgatgcg ttaaaagcgg cagatcaaca cgcgaaagcg gaagctatcg
     4921 gtcaggccga tatgttcggc gtgctggccg aagagccgga acaaattgaa caatcctacg
     4981 ccagctgcca accgtggccg gagcaggtgg tattagatgg ggaacgtgaa acgttaggcc
     5041 tgtacctgac cggacaccct atcaaccagt atttaaaaga gattgagcgt tatgtcggag
     5101 gcgtaaggct gaaagacatg cacccgacag aacgtggtaa agtcatcacg gctgcggggc
     5161 tcgttgttgc cgcgcgggtt atggtcacca agcgcggcaa tcgtatcggt atctgcacgc
     5221 tggatgaccg ttccgggcgg ctggaagtga tgttgtttac tgacgccctg gataaatacc
     5281 agcaattgct ggaaaaagac cgcatactta tcgtcagcgg acaggtcagc tttgatgact
     5341 tcagcggtgg gcttaaaatg accgctcgcg aagtgatgga tattgacgaa gcccgggaaa
     5401 aatatgctcg cgggcttgct atctcgctga cggacaggca aattgatgac cagcttttaa
     5461 accgactccg tcagtctctg gaaccccacc gctctgggac aattccagta catctctact
     5521 atcagagggc ggatgcacgc gcgcggttgc gttttggcgc gacgtggcgt gtctctccga
     5581 gcgatcgttt attaaacgat ctccgtggcc tcattggttc ggagcaggtg gaactggagt
     5641 ttgactaaac ttaattaacg gcactcctca gccaagtcaa aagcctccga ccggaggctt
     5701 ttgactacat gcccatggcg tttacgcccc gccctgccac tcatcgcagt actgttgtaa
     5761 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
     5821 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatagtga aaacgggggc
     5881 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
     5941 ggctgagacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
     6001 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
     6061 actccagagc gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
     6121 actatcccat atcaccagct caccgtcttt cattgccata cggaactccg gatgagcatt
     6181 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
     6241 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
     6301 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
     6361 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
     6421 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgcca
     6481 agccaaatag gccgt
//
