LOCUS       sequence-123373-        4944 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 4944)
  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 4944)
  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-C3
FEATURES             Location/Qualifiers
     source          1..4944
                     /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      4124..4155
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4171..4830)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4745..4764
                     /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 ttatcaggtc ttagcccgaa aatggcgccc acaaaccttt gctgacgtcg
     2221 tcggccagga acatgtgctg accgcactgg cgaacggctt gtcgttaggg cgtattcatc
     2281 atgcttatct tttttccggc acccgtggcg tcggaaaaac ctctatcgcc cgactgctgg
     2341 cgaaggggct aaactgcgaa accggcatta ccgcgacgcc gtgcggcgtg tgcgataact
     2401 gtcgtgaaat cgagcagggg cgctttgtcg atctgattga aatcgacgcc gcctcgcgca
     2461 ccaaagttga agatacccgc gacctgctgg ataacgtcca gtacgctccg gcgcgtgatc
     2521 gtttcaaagt ttatctgatc gacgaagtgc atatgctgtc gcgccacagc tttaacgcac
     2581 tgttaaaaac ccttgaagag ccgccggagc acgttaagtt tctgctggcg acgaccgatc
     2641 cacagaaatt gccggtgacg attttgtcac gctgtctgca atttcatctc aaggcgctgg
     2701 atgtcgagca aattcgccat cagcttgagc acatcctcaa cgaagaacat atcgctcacg
     2761 agccgcgggc gctgcaattg ctggcacgcg ccgctgaagg cagcctgcga gatgccttaa
     2821 gtctgaccga ccaggcgatt gccagcggtg acggccaggt ttcaacccag gcggtcagtg
     2881 cgatgctggg tacgcttgac gacgatcagg cgctgtcgct ggttgaagcg atggtcgagg
     2941 ccaacggcga gcgcgtaatg gcgctgatta atgaagccgc tgcccgtggt atcgagtggg
     3001 aagcgttgct ggtggaaatg ctcggcctgt tgcatcgtat tgcgatggta caactttcgc
     3061 ctgctgcact tggcaacgac atggccgcca tcgagctgcg gatgcgtgaa ctggcgcgca
     3121 ccataccgcc gacggatatt cagctttact atcagacgct gttgattggt cgcaaagaat
     3181 taccgtatgc gccggaccgt cgcatgggcg ttgagatgac gctgctgcgc gcgctggcat
     3241 tccatccgcg tatgccgctg cctgagccag aagtgccacg acagtccttt gcacccgtcg
     3301 cgccaacggc agtaatgacg ccaacccagg tgccgccgca accgcaatca gcgccgcagc
     3361 aggcaccgac tgtaccgctc ccggaaacca ccagccaggt gctggcggcg cgccagcagt
     3421 tgcagcgcgt gcagggagca accaaagcaa aaaagagtga accggcagcc gctacccgcg
     3481 cgcggccggt gaataacgct gcgctggaaa gactggcttc ggtcaccgat cgcgttcagg
     3541 cgcgtccggt gccatcggcg ctggaaaaag cgccagccaa aaaagaagcg tatcgctgga
     3601 aggcgaccac tccggtgatg cagcaaaaag aagtggtcgc cacgccgaag gcgctgaaaa
     3661 aagcgctgga acatgaaaaa acgccggaac tggcggcgaa gctagcggca gaagccattg
     3721 agcgcgaccc gtgggcggca caggtgagcc aactttcgct accaaaactg gtcgaacagg
     3781 tggcgttaaa tgcctggaaa gaggagagcg acaacgcagt atgtctgcat ttgcgctcct
     3841 ctcagcggca tttgaacaac cgcggtgcac agcaaaaact ggctgaagcg ttgagcatgt
     3901 taaaaggttc aacggttgaa ctgactatcg ttgaagatga taatcccgcg gtgcgtacgc
     3961 cgctggagtg gcgtcaggcg atatacgaag aaaaacttgc gcaggcgcgc gagtccatta
     4021 ttgcggataa taatattcag accctgcgtc ggttcttcga tgcggagctg gatgaagaaa
     4081 gtatccgccc catttaaact taattaacgg cactcctcag ccaagtcaaa agcctccgac
     4141 cggaggcttt tgactacatg cccatggcgt ttacgccccg ccctgccact catcgcagta
     4201 ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacaaacgg catgatgaac
     4261 ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc ccatagtgaa
     4321 aacgggggcg aagaagttgt ccatattggc cacgtttaaa tcaaaactgg tgaaactcac
     4381 ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga aataggccag
     4441 gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc ggaaatcgtc
     4501 gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa cggtgtaaca
     4561 agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac ggaactccgg
     4621 atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt
     4681 tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca
     4741 ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac
     4801 ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga
     4861 taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
     4921 tacgtgccaa gccaaatagg ccgt
//
