LOCUS       sequence-123439-        6665 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 6665)
  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 6665)
  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-U
FEATURES             Location/Qualifiers
     source          1..6665
                     /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             4908..5162
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5192..5818
                     /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(5845..5876)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(5892..6551)
                     /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 gttttttggg ctaacaggag gaattaacca tgagcactgc
     2161 aattacacgc cagatcgttc tcgctaccgc aaccaccggt atgaaccaga ttggtgcgca
     2221 ctatgaaggc cacaagatca ttgagattgg tgccgttgaa gtggtgaacc gtcgcctgac
     2281 gggcaataac ttccatgttt atctcaaacc cgatcggctg gtggatccgg aagcctttgg
     2341 cgtacatggt attgccgatg aatttttgct cgataagccc acgtttgccg aagtagccga
     2401 tgagttcatg gactatattc gcggcgcgga gttggtgatc cataacgcag cgttcgatat
     2461 cggctttatg gactacgagt tttcgttgct taagcgcgat attccgaaga ccaatacttt
     2521 ctgtaaggtc accgatagcc ttgcggtggc gaggaaaatg tttcccggta agcgcaacag
     2581 cctcgatgcg ttatgtgctc gctacgaaat agataacagt aaacgaacgc tgcacggggc
     2641 attactcgat gcccagatcc ttgcggaagt ttatctggcg atgaccggtg gtcaaacgtc
     2701 gatggctttt gcgatggaag gagagacaca acagcaacaa ggtgaagcaa caattcagcg
     2761 cattgtacgt caggcaagta agttacgcgt tgtttttgcg acagatgaag agattgcagc
     2821 tcatgaagcc cgtctcgatc tggtgcagaa gaaaggcgga agttgcctct ggcgagcata
     2881 atttaatatc agtaaaccgg acataaccca tgaagaaaaa tcgcgctttt ttgaagtggg
     2941 cagggggcaa gtatcccctg cttgatgata ttaaacggca tttgcccaag ggcgaatgtc
     3001 tggttgagcc ttttgtaggt gccgggtcgg tgtttctcaa caccgacttt tctcgttata
     3061 tccttgccga tatcaatagc gacctgatca gtctctataa cattgtgaag atgcgtactg
     3121 atgagtacgt acaggccgca cgcgagctgt ttgttcccga aacaaattgc gccgaggttt
     3181 actatcagtt ccgcgaagag ttcaacaaaa gccaggatcc gttccgtcgg gcggtactgt
     3241 ttttatattt gaaccgctac ggttacaacg gcctgtgtcg ttacaatctg cgcggtgagt
     3301 ttaacgtgcc gttcggccgc tacaaaaaac cctatttccc ggaagcagag ttgtatcact
     3361 tcgctgaaaa agcgcagaat gcctttttct attgtgagtc ttacgccgat agcatggcgc
     3421 gcgcagatga tgcatccgtc gtctattgcg atccgcctta tgcaccgctg tctgcgaccg
     3481 ccaactttac ggcgtatcac acaaacagtt ttacgcttga acaacaagcg catctggcgg
     3541 agatcgccga aggtctggtt gagcgccata ttccagtgct gatctccaat cacgatacga
     3601 tgttaacgcg tgagtggtat cagcgcgcaa aattgcatgt cgtcaaagtt cgacgcagta
     3661 taagcagcaa cggcggcaca cgtaaaaagg tggacgaact gctggctttg tacaaaccag
     3721 gagtcgtttc acccgcgaaa aaataattca gctaagacac tgcactggat taagatgaaa
     3781 acgattgaag ttgatgatga actctacagc tatattgcca gccacactaa gcatatcggc
     3841 gagagcgcat ccgacatttt acggcgtatg ttgaaatttt ccgccgcatc acagcctgct
     3901 gctccggtga cgaaagaggt tcgcgttgcg tcacctgcta tcgtcgaagc gaagccggtc
     3961 aaaacgatta aagacaaggt tcgcgcaatg cgtgaacttc tgctttcgga tgaatacgca
     4021 gagcaaaagc gagcggtcaa tcgctttatg ctgctgttgt ctacactata ttctcttgac
     4081 gcccaggcgt ttgccgaagc aacggaatcg ttgcacggtc gtacacgcgt ttactttgcg
     4141 gcagatgaac aaacgctgct gaaaaatggt aatcagacca agccgaaaca tgtgccaggc
     4201 acgccgtatt gggtgatcac caacaccaac accggccgta aatgcagcat gatcgaacac
     4261 atcatgcagt cgatgcaatt cccggcggaa ttgattgaga aggtttgcgg aactatctaa
     4321 cggctgaaat taatgaggtc atacccaaat ggatagttcg tttacgccca ttgaacaaat
     4381 gctaaaattt cgcgccagcc gccacgaaga ttttccttat caggagatcc ttctgactcg
     4441 tctttgcatg cacatgcaaa gcaagctgct ggagaaccgc aataaaatgc tgaaggctca
     4501 gggaattaac gagacgttgt ttatggcgtt gattacgctg gagtctcagg aaaaccacag
     4561 tattcagcct tctgaattaa gttgtgctct tggatcatcc cgtaccaacg cgacgcgtat
     4621 tgccgatgaa ctggaaaaac gcggttggat cgaacgtcgt gaaagcgata acgatcgccg
     4681 ctgcctgcat ctgcaattaa cggaaaaagg tcacgagttt ttgcgcgagg ttttaccacc
     4741 gcagcataac tgcctgcatc aactctggtc cgcgctcagc acaacagaaa aagatcagct
     4801 cgagcaaatc acccgcaaat tgctctcccg tctcgaccag atggaacaag acggtgtggt
     4861 tctcgaagcg atgagctaat aatacaaaaa ttaggaggaa tttcaacatg acaaatttat
     4921 ctgacatcat tgaaaaagaa acaggaaaac aactagtgat tcaagaatca attctaatgt
     4981 taccagaaga agtagaggaa gtaattggga ataaaccaga aagtgatatt ttagttcata
     5041 ctgcttatga tgaaagtaca gatgaaaatg taatgctatt aacttcagat gctccagaat
     5101 ataaaccttg ggctttagta attcaagaca gtaatggaga aaataaaatt aaaatgttat
     5161 aagtcgagat taagtaaacc ggaatctgaa gatgaccgac gcggaatacg ttcgtatcca
     5221 cgaaaaactg gacatctaca ccttcaaaaa acagttcttc aacaacaaaa aatctgtttc
     5281 tcaccgttgc tacgttctgt tcgaactgaa acgtcgtggt gaacgtcgtg cgtgcttctg
     5341 gggttacgcg gttaacaaac cgcagtctgg taccgaacgt ggtatccacg cggaaatctt
     5401 ctctatccgt aaagttgaag aatacctgcg tgacaacccg ggtcagttca ccatcaactg
     5461 gtactcttct tggtctccgt gcgcggactg cgcggaaaaa atcctggaat ggtacaacca
     5521 ggaactgcgt ggtaacggtc acaccctgaa aatctgggcg tgcaaactgt actacgaaaa
     5581 aaacgcgcgt aaccagatcg gtctgtggaa cctgcgtgac aacggtgttg gtctgaacgt
     5641 tatggtttct gaacactacc agtgctgccg taaaatcttc atccagtctt ctcacaacca
     5701 gctgaacgaa aaccgttggc tggaaaaaac cctgaaacgt gcggaaaaac gtcgttctga
     5761 actgtctatc atgatccagg ttaaaatcct gcacaccacc aaatctccgg cggtttaaac
     5821 ttaattaacg gcactcctca gccaagtcaa aagcctccgg tcggaggctt ttgactacat
     5881 gcccatggcg tttacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     5941 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
     6001 agcaccttgt cgccttgcgt ataatatttg cccatagtga aaacgggggc gaagaagttg
     6061 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
     6121 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     6181 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     6241 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     6301 atcaccagct caccgtcttt cattgccata cggaactccg gatgagcatt catcaggcgg
     6361 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     6421 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagtaac tgactgaaat
     6481 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     6541 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     6601 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgcca agccaaatag
     6661 gccgt
//
