LOCUS       sequence-446226-        8705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Untargetted bacterial drift.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8705)
  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 8705)
  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: DP6
FEATURES             Location/Qualifiers
     source          1..8705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..879
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     primer_bind     complement(929..946)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1099..1185
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1277..1304
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     protein_bind    1468..1486
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1556..2830
                     /codon_start=1
                     /gene="III"
                     /product="pIII"
                     /label=M13 gene III
                     /note="minor coat protein"
                     /translation="MKKLLFAIPLVVPFYSHSAETVESCLAKPHTENSFTNVWKDDKTL
                     DRYANYEGCLWNATGVVVCTGDETQCYGTWVPIGLAIPENEGGGSEGGGSEGGGSEGGG
                     TKPPEYGDTPIPGYTYINPLDGTYPPGTEQNPANPNPSLEESQPLNTFMFQNNRFRNRQ
                     GALTVYTGTVTQGTDPVKTYYQYTPVSSKAMYDAYWNGKFRDCAFHSGFNEDPFVCEYQ
                     GQSSDLPQPPVNAGGGSGGGSGGGSEGGGSEGGGSEGGGSEGGGSGGGSGSGDFDYEKM
                     ANANKGAMTENADENALQSDAKGKLDSVATDYGAAIDGFIGDVSGLANGNGATGDFAGS
                     NSQMAQVGDGDNSPLMNNFRQYLPSLPQSVECRPFVFGAGKPYEFSIDCDKINLFRGVF
                     AFLLYVATFMYVFSTFANILRNKES"
     CDS             3943..4602
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAKCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4009..4028)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      complement(4618..4649)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     CDS             complement(4676..5302)
                     /codon_start=1
                     /product="cytidine deaminase from the sea lamprey 
                     Petromyzon marinus (Rogozin et al., 2007)"
                     /label=PmCDA1
                     /translation="MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER
                     RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKI
                     LEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIF
                     IQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
     CDS             complement(5332..5586)
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
                     AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     promoter        complement(8395..8679)
                     /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     complement(8453..8472)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 atggctgaag cgcaaaatga tcccctgctg ccgggatact cgtttaatgc ccatctggtg
       61 gcgggtttaa cgccgattga ggccaacggt tatctcgatt tttttatcga ccgaccgctg
      121 ggaatgaaag gttatattct caatctcacc attcgcggtc agggggtggt gaaaaatcag
      181 ggacgagaat ttgtttgccg accgggtgat attttgctgt tcccgccagg agagattcat
      241 cactacggtc gtcatccgga ggctcgcgaa tggtatcacc agtgggttta ctttcgtccg
      301 cgcgcctact ggcatgaatg gcttaactgg ccgtcaatat ttgccaatac ggggttcttt
      361 cgcccggatg aagcgcacca gccgcatttc agcgacctgt ttgggcaaat cattaacgcc
      421 gggcaagggg aagggcgcta ttcggagctg ctggcgataa atctgcttga gcaattgtta
      481 ctgcggcgca tggaagcgat taacgagtcg ctccatccac cgatggataa tcgggtacgc
      541 gaggcttgtc agtacatcag cgatcacctg gcagacagca attttgatat cgccagcgtc
      601 gcacagcatg tttgcttgtc gccgtcgcgt ctgtcacatc ttttccgcca gcagttaggg
      661 attagcgtct taagctggcg cgaggaccaa cgtatcagcc aggcgaagct gcttttgagc
      721 accacccgga tgcctatcgc caccgtcggt cgcaatgttg gttttgacga tcaactctat
      781 ttctcgcggg tatttaaaaa atgcaccggg gccagcccga gcgagttccg tgccggttgt
      841 gaagaaaaag tgaatgatgt agccgtcaag ttgtcataaa cagtcagtag ggccctggct
      901 gttttggcgg atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg
      961 gtctgataaa acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc
     1021 cgaactcaga agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag
     1081 tagggaactg ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     1141 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat
     1201 ttgaacgttg cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc
     1261 aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact
     1321 cgctgaggac attggctccg tgatgaaatt cgccacttgt tagtgtaatt cgctaactca
     1381 tcctggcatg ttgctgttga ttcttcaatc agatctttat aaatcaaaaa gataaaaaat
     1441 tggcacgcaa attgtattaa cagttcatcc ctatcagtga tagagagcag gacaatcctg
     1501 aacgcagaaa tcaagaggac aacattacct gcaggtgcag taaaggaaaa aaaaaatgaa
     1561 aaaattatta ttcgcaattc ctttagttgt tcctttctat tctcactccg ctgaaactgt
     1621 tgaaagttgt ttagcaaaac cccatacaga aaattcattt actaacgtct ggaaagacga
     1681 caaaacttta gatcgttacg ctaactatga gggctgtctg tggaatgcta caggcgttgt
     1741 agtttgtact ggtgacgaaa ctcagtgtta cggtacatgg gttcctattg ggcttgctat
     1801 ccctgaaaat gagggtggtg gctctgaggg tggcggttct gagggtggcg gttctgaggg
     1861 tggcggtact aaacctcctg agtacggtga tacacctatt ccgggctata cttatatcaa
     1921 ccctctcgac ggcacttatc cgcctggtac tgagcaaaac cccgctaatc ctaatccttc
     1981 tcttgaggag tctcagcctc ttaatacttt catgtttcag aataataggt tccgaaatag
     2041 gcagggggca ttaactgttt atacgggcac tgttactcaa ggcactgacc ccgttaaaac
     2101 ttattaccag tacactcctg tatcatcaaa agccatgtat gacgcttact ggaacggtaa
     2161 attcagagac tgcgctttcc attctggctt taatgaggat ccattcgttt gtgaatatca
     2221 aggccaatcg tctgacctgc ctcaacctcc tgtcaatgct ggcggcggct ctggtggtgg
     2281 ttctggtggc ggctctgagg gtggtggctc tgagggtggc ggttctgagg gtggcggctc
     2341 tgagggaggc ggttccggtg gtggctctgg ttccggtgat tttgattatg aaaagatggc
     2401 aaacgctaat aagggggcta tgaccgaaaa tgccgatgaa aacgcgctac agtctgacgc
     2461 taaaggcaaa cttgattctg tcgctactga ttacggtgct gctatcgatg gtttcattgg
     2521 tgacgtttcc ggccttgcta atggtaatgg tgctactggt gattttgctg gctctaattc
     2581 ccaaatggct caagtcggtg acggtgataa ttcaccttta atgaataatt tccgtcaata
     2641 tttaccttcc ctccctcaat cggttgaatg tcgccctttt gtctttggcg ctggtaaacc
     2701 ttacgagttc agtatcgact gcgataagat caacctgttc cgcggtgtct ttgcgtttct
     2761 tttatatgtt gccaccttta tgtatgtatt ttctacgttt gctaacatac tgcgtaataa
     2821 ggagtcttaa agggtcccta agtctcctca gcaaaacgaa aggcccagtc tttcgactga
     2881 gcctttcgtt ttatttgacc ggatgtcctc ttcctcctgg gctgacttca ggtgctacat
     2941 ttgaagagat aaattgcact gaaatctaga gcggttcagt agaaaagatc aaaggatctt
     3001 cttgagatcc tttttttctg cgcgtaatct tttgccctgt aaacgaaaaa accacctggg
     3061 gaggtggttt gatcgaaggt taagcagttc accaactgac ttaaccttcg aaaaaaccac
     3121 ctggggaggt ggtttgatcg aaggttaagt cagttgggga actgcttaac cgtggtaact
     3181 ggctttcgca gagcacagca accaaatctg tccttccagt gtagccggac tttggcgcac
     3241 acttcaagag caaccgcgtg tttagctaaa caaatcctct gcgaactccc agttaccaat
     3301 ggctgctgcc agtggcgttt taccgtgctt ttccgggttg gactcaagtg aacagttacc
     3361 ggataaggcg cagcagtcgg gctgaacggg gagttcttgc ttacagccca gcttggagcg
     3421 aacgacctac accgagccga gataccagtg tgtgagctat gagaaagcgc cacacttccc
     3481 gtaagggaga aaggcggaac aggtatccgg taaacggcag ggtcggaaca ggagagcgca
     3541 agagggagcg acccgccgga aacggtgggg atctttaagt cctgtcgggt ttcgcccgta
     3601 ctgtcagatt catggttgag cctcacggct cccacagatg caccggaaaa gcgtctgttt
     3661 atgtgaactc tggcaggagg gcggagccta tggaaaaacg ccaccggcgc ggccctgctg
     3721 ttttgcctca catgttagtc ccctgcttat ccacggaatc tgtgggtaac tttgtatgtg
     3781 tccgcagcgc ccgccgcagt ctcacgcccg gagcgtagcg accgagtgac ggcctatttg
     3841 gcttggcacg taagaggttc caactttcac cataatgaaa taagatcact accgggcgta
     3901 ttttttgagt tatcgagatt ttcaggagct aaggaagcta aaatggagaa aaaaatcact
     3961 ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga ggcatttcag
     4021 tcagttgcta aatgtaccta taaccagacc gttcagctgg atattacggc ctttttaaag
     4081 accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct tgcccgcctg
     4141 atgaatgctc atccggagtt ccgtatggca atgaaagacg gtgagctggt gatatgggat
     4201 agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc atcgctctgg
     4261 agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga tgtggcgtgt
     4321 tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt tttcgtctca
     4381 gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat ggacaacttc
     4441 ttcgcccccg ttttcactat gggcaaatat tatacgcaag gcgacaaggt gctgatgccg
     4501 ctggcgattc aggttcatca tgccgtttgt gatggcttcc atgtcggcag aatgcttaat
     4561 gaattacaac agtactgcga tgagtggcag ggcggggcgt aaacgccatg ggcatgtagt
     4621 caaaagcctc cggtcggagg cttttgactt ggctgaggag tgccgttaat taagtttaaa
     4681 ccgccggaga tttggtggtg tgcaggattt taacctggat catgatagac agttcagaac
     4741 gacgtttttc cgcacgtttc agggtttttt ccagccaacg gttttcgttc agctggttgt
     4801 gagaagactg gatgaagatt ttacggcagc actggtagtg ttcagaaacc ataacgttca
     4861 gaccaacacc gttgtcacgc aggttccaca gaccgatctg gttacgcgcg tttttttcgt
     4921 agtacagttt gcacgcccag attttcaggg tgtgaccgtt accacgcagt tcctggttgt
     4981 accattccag gattttttcc gcgcagtccg cgcacggaga ccaagaagag taccagttga
     5041 tggtgaactg acccgggttg tcacgcaggt attcttcaac tttacggata gagaagattt
     5101 ccgcgtggat accacgttcg gtaccagact gcggtttgtt aaccgcgtaa ccccagaagc
     5161 acgcacgacg ttcaccacga cgtttcagtt cgaacagaac gtagcaacgg tgagaaacag
     5221 attttttgtt gttgaagaac tgttttttga aggtgtagat gtccagtttt tcgtggatac
     5281 gaacgtattc cgcgtcggtc atcttcagat tccggtttac ttaatctcga cttataacat
     5341 tttaatttta ttttctccat tactgtcttg aattactaaa gcccaaggtt tatattctgg
     5401 agcatctgaa gttaatagca ttacattttc atctgtactt tcatcataag cagtatgaac
     5461 taaaatatca ctttctggtt tattcccaat tacttcctct acttcttctg gtaacattag
     5521 aattgattct tgaatcacta gttgttttcc tgtttctttt tcaatgatgt cagataaatt
     5581 tgtcatgttg aaattcctcc taatttttgt attattagct catcgcttcg agaaccacac
     5641 cgtcttgttc catctggtcg agacgggaga gcaatttgcg ggtgatttgc tcgagctgat
     5701 ctttttctgt tgtgctgagc gcggaccaga gttgatgcag gcagttatgc tgcggtggta
     5761 aaacctcgcg caaaaactcg tgaccttttt ccgttaattg cagatgcagg cagcggcgat
     5821 cgttatcgct ttcacgacgt tcgatccaac cgcgtttttc cagttcatcg gcaatacgcg
     5881 tcgcgttggt acgggatgat ccaagagcac aacttaattc agaaggctga atactgtggt
     5941 tttcctgaga ctccagcgta atcaacgcca taaacaacgt ctcgttaatt ccctgagcct
     6001 tcagcatttt attgcggttc tccagcagct tgctttgcat gtgcatgcaa agacgagtca
     6061 gaaggatctc ctgataagga aaatcttcgt ggcggctggc gcgaaatttt agcatttgtt
     6121 caatgggcgt aaacgaacta tccatttggg tatgacctca ttaatttcag ccgttagata
     6181 gttccgcaaa ccttctcaat caattccgcc gggaattgca tcgactgcat gatgtgttcg
     6241 atcatgctgc atttacggcc ggtgttggtg ttggtgatca cccaatacgg cgtgcctggc
     6301 acatgtttcg gcttggtctg attaccattt ttcagcagcg tttgttcatc tgccgcaaag
     6361 taaacgcgtg tacgaccgtg caacgattcc gttgcttcgg caaacgcctg ggcgtcaaga
     6421 gaatatagtg tagacaacag cagcataaag cgattgaccg ctcgcttttg ctctgcgtat
     6481 tcatccgaaa gcagaagttc acgcattgcg cgaaccttgt ctttaatcgt tttgaccggc
     6541 ttcgcttcga cgatagcagg tgacgcaacg cgaacctctt tcgtcaccgg agcagcaggc
     6601 tgtgatgcgg cggaaaattt caacatacgc cgtaaaatgt cggatgcgct ctcgccgata
     6661 tgcttagtgt ggctggcaat atagctgtag agttcatcat caacttcaat cgttttcatc
     6721 ttaatccagt gcagtgtctt agctgaatta ttttttcgcg ggtgaaacga ctcctggttt
     6781 gtacaaagcc agcagttcgt ccaccttttt acgtgtgccg ccgttgctgc ttatactgcg
     6841 tcgaactttg acgacatgca attttgcgcg ctgataccac tcacgcgtta acatcgtatc
     6901 gtgattggag atcagcactg gaatatggcg ctcaaccaga ccttcggcga tctccgccag
     6961 atgcgcttgt tgttcaagcg taaaactgtt tgtgtgatac gccgtaaagt tggcggtcgc
     7021 agacagcggt gcataaggcg gatcgcaata gacgacggat gcatcatctg cgcgcgccat
     7081 gctatcggcg taagactcac aatagaaaaa ggcattctgc gctttttcag cgaagtgata
     7141 caactctgct tccgggaaat agggtttttt gtagcggccg aacggcacgt taaactcacc
     7201 gcgcagattg taacgacaca ggccgttgta accgtagcgg ttcaaatata aaaacagtac
     7261 cgcccgacgg aacggatcct ggcttttgtt gaactcttcg cggaactgat agtaaacctc
     7321 ggcgcaattt gtttcgggaa caaacagctc gcgtgcggcc tgtacgtact catcagtacg
     7381 catcttcaca atgttataga gactgatcag gtcgctattg atatcggcaa ggatataacg
     7441 agaaaagtcg gtgttgagaa acaccgaccc ggcacctaca aaaggctcaa ccagacattc
     7501 gcccttgggc aaatgccgtt taatatcatc aagcagggga tacttgcccc cctgcccact
     7561 tcaaaaaagc gcgatttttc ttcatgggtt atgtccggtt tactgatatt aaattatgct
     7621 cgccagaggc aacttccgcc tttcttctgc accagatcga gacgggcttc atgagctgca
     7681 atctcttcat ctgtcgcaaa aacaacgcgt aacttacttg cctgacgtac aatgcgctga
     7741 attgttgctt caccttgttg ctgttgtgtc tctccttcca tcgcaaaagc catcgacgtt
     7801 tgaccaccgg tcatcgccag ataaacttcc gcaaggatct gggcatcgag taatgccccg
     7861 tgcagcgttc gtttactgtt atctatttcg tagcgagcac ataacgcatc gaggctgttg
     7921 cgcttaccgg gaaacatttt cctcgccacc gcaaggctat cggtgacctt acagaaagta
     7981 ttggtcttcg gaatatcgcg cttaagcaac gaaaactcgt agtccataaa gccgatatcg
     8041 aacgctgcgt tatggatcac caactccgcg ccgcgaatat agtccatgaa ctcatcggct
     8101 acttcggcaa acgtgggctt atcgagcaaa aattcatcgg caataccatg tacgccaaag
     8161 gcttccggat ccaccagccg atcgggtttg agataaacat ggaagttatt gcccgtcagg
     8221 cgacggttca ccacttcaac ggcaccaatc tcaatgatct tgtggccttc atagtgcgca
     8281 ccaatctggt tcataccggt ggttgcggta gcgagaacga tctggcgtgt aattgcagtg
     8341 ctcatagcgg tcatttatgt cagacttgag ttgtacgcgt ccaaaaaaac gggtatggag
     8401 aaacagtaga gagttgcgat aaaaagcgtc aggtaggatc cgctaatctt atggataaaa
     8461 atgctatggc atagcaaagt gtgacgccgt gcaaataatc aatgtggact tttctgccgt
     8521 gattatagac acttttgtta cgcgtttttg tcatggcttt ggtcccgctt tgttacagaa
     8581 tgcttttaat aagcggggtt accggttggg ttagcgagaa gagccagtaa aagacgcagt
     8641 gacggcaatg tctgatgcaa tatggacaat tggtttcttc tctgaatggt gggagtatga
     8701 aaagt
//
