LOCUS       sequence-123425-        6185 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 6185)
  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 6185)
  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-R2
FEATURES             Location/Qualifiers
     source          1..6185
                     /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             4365..4619
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             4649..5338
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
                     GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
                     SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
                     EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
                     LWATGLK"
     terminator      5365..5396
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(5412..6071)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
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 accagatcgg tgcgcactat gaaggccaca agatcattga gattggtgcc gttgaagtgg
     2281 tgaaccgtcg cctgacgggc aacaacttcc 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 agcataattc aacatcagta aaccggacat aacccatgaa gaaaaatcgc
     2941 gcttttttga agtgggcagg gggcaagtat cccctgcttg atgatattaa acggcatttg
     3001 cccaagggcg aatgtctggt tgagcctttt gtaggtgccg ggtcggtgtt tctcaacacc
     3061 gacttttctc gttacatcct tgccgatatc aatagcgacc tgatcagtct ctataacatt
     3121 gtgaagatgc gtactgatga gtacgtacag gccgcacgcg agctgtttgt tcccgaaaca
     3181 aattgcgccg aggtttacta tcagttccgc gaagagttca acaaaagcca ggatccgttc
     3241 cgtcgggcgg tactgttctt atatttgaac cgctacggtt acaacggcct gtgtcgttac
     3301 aatctgcgcg gtgagtttaa cgtgccgttc ggccgctaca aaaaacccta tttcccggaa
     3361 gcagagttgt atcacttcgc tgaaaaagcg cagaacgcct ttttctattg tgagtcttac
     3421 gccgacagca tggcgcgcgc agatgatgca tccgtcgtct attgcgatcc gccttatgca
     3481 ccgctgtctg cgaccgccaa ctttacggcg tatcacacaa acagttttac gcttgaacaa
     3541 caagcgcatc tggcggagat cgccgaaggt ctggttgagc gccatattcc agtgctgatc
     3601 tccaatcacg atacgatgtt aacgcgtgag tggtatcagc gcgcgaagct gcatgtcgtc
     3661 aaagttcgac gcagcataag cagcaacggc ggcacacgta aaaaggtgga cgaactgctg
     3721 gctttgtaca aaccaggagt cgtttcaccc gcgaaaaaat aattcagcta agacactgca
     3781 ctggattaag atgaaaacga tcgaagttga tgacgaactc tacagctata tcgccagcca
     3841 cactaagcat atcggcgaga gcgcatccga cattttacgg cgtatgttga aattttccgc
     3901 cgcatcacag cctgctgctc cggtgacgaa agaggttcgc gttgcgtcac ctgctatcgt
     3961 cgaagcgaag ccggtcaaaa cgattaaaga caaggttcgc gcaatgcgtg aacttctgct
     4021 ttcggatgaa tacgcagagc aaaagcgagc ggtcaatcgc tttatgctgc tgttgtctac
     4081 actatattct cttgacgccc aggcgtttgc cgaagcaacg gaatcgttgc acggtcgtac
     4141 acgcgtttac tttgcggcag atgaacaaac gctgctgaaa aacggtaatc agaccaagcc
     4201 gaaacatgtg ccaggcacgc cgtattgggt gatcaccaac accaacaccg gccgtaaatg
     4261 cagcatgatc gaacacatca tgcagtcgat gcaattcccg gcggaactga ttgagaaggt
     4321 ttgcggaact atctaataat acaaaaacta ggaggaactt caacatgaca aatttatctg
     4381 acatcattga aaaagaaaca ggaaaacaac tagtgattca agaatcaatt ctaatgttac
     4441 cagaagaagt agaggaagta atcgggaata aaccagaaag tgatatttta gttcatactg
     4501 cttatgacga aagtacagat gaaaacgtaa tgctattaac ttcagatgct ccagaataca
     4561 aaccttgggc gttagtaatt caagacagta atggagaaaa caagattaaa atgttgtaag
     4621 tcgagattaa gtaaaccgga atctgaagat gtcttctgaa accggtccgg ttgcggttga
     4681 cccgaccctg cgtcgtcgta tcgaaccgca cgaattcgaa gttttcttcg acccgcgtga
     4741 actgcgtaaa gaaacctgcc tgctgtacga aatcaactgg ggtggtcgtc actctatctg
     4801 gcgtcacacc tctcagaaca ccaacaaaca cgttgaagtt aacttcatcg aaaaattcac
     4861 caccgaacgt tacttctgcc cgaacacccg ttgctctatc acctggttcc tgtcttggtc
     4921 tccgtgcggt gaatgctctc gtgcgatcac cgaattcctg tctcgttacc cgcacgttac
     4981 cctgttcatc tacatcgcgc gtctgtacca ccacgcggac ccgcgtaacc gtcagggtct
     5041 gcgtgacctg atctcttctg gtgtgaccat ccagatcatg accgaacagg aatctggtta
     5101 ctgctggcgt aacttcgtta actactctcc gtctaacgaa gcgcactggc cgcgttaccc
     5161 gcacctgtgg gttcgtctgt acgttctgga actgtactgc atcatcctgg gtctgccgcc
     5221 gtgcctgaac atcctgcgtc gtaaacagcc gcagctgacc ttcttcacca tcgcgctgca
     5281 gtcttgccac taccagcgtc tgccgccgca catcctgtgg gcgaccggtc tgaaataaac
     5341 ttaattaacg gcactcctca gccaagtcaa aagcctccga ccggaggctt ttgactacat
     5401 gcccatggcg tttacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     5461 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
     5521 agcaccttgt cgccttgcgt ataatatttg cccatagtga aaacgggggc gaagaagttg
     5581 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
     5641 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     5701 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     5761 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     5821 atcaccagct caccgtcttt cattgccata cggaactccg gatgagcatt catcaggcgg
     5881 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     5941 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagtaac tgactgaaat
     6001 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     6061 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     6121 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgcca agccaaatag
     6181 gccgt
//
