LOCUS       V007025                 8639 bp    DNA     circular SYN 13-MAY-2021
DEFINITION  Exported.
ACCESSION   V007025
VERSION     V007025
KEYWORDS    DP6.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8639)
  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 8639)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8639)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat
            Commun."; date: "2015-10-7"; pages: "
            10.1038/ncomms9425"
            SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..876
                     /label="araC"
                     /note="L-arabinose regulatory protein"
     primer_bind     complement(929..946)
                     /label="pBAD Reverse"
                     /note="For vectors with E. coli araBAD promoter, reverse
                     primer"
     terminator      1099..1185
                     /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
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1556..2827
                     /label="M13 gene III"
                     /note="pIII"
     rep_origin      2987..3725
                     /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."
     CDS             3878..4534
                     /label="CmR"
                     /note="chloramphenicol acetyltransferase"
     terminator      complement(4553..4584)
                     /label="tonB terminator"
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     CDS             complement(4614..5237)
                     /label="PmCDA1"
                     /note="cytidine deaminase from the sea lamprey Petromyzon
                     marinus (Rogozin et al., 2007)"
     CDS             complement(5270..5521)
                     /label="UGI"
                     /note="uracil-DNA glycosylase inhibitor from a Bacillus
                     subtilis bacteriophage (Mol et al., 1995)"
     CDS             complement(5553..6080)
                     /gene="mprA"
                     /label="Transcriptional repressor MprA"
                     /note="Transcriptional repressor MprA from Escherichia coli
                     (strain K12). Accession#: P0ACR9"
     CDS             complement(6112..6654)
                     /gene="seqA"
                     /label="Negative modulator of initiation of replication"
                     /note="Negative modulator of initiation of replication from
                     Escherichia coli (strain K12). Accession#: P0AFY8"
     CDS             complement(6686..7519)
                     /gene="dam"
                     /label="DNA adenine methylase"
                     /note="DNA adenine methylase from Escherichia coli (strain
                     K12). Accession#: P0AEE8"
     CDS             complement(7551..8279)
                     /gene="dnaQ"
                     /label="DNA polymerase III subunit epsilon"
                     /note="DNA polymerase III subunit epsilon from Escherichia
                     coli (strain K12). Accession#: P03007"
     promoter        complement(8329..8613)
                     /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)"
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 taagtcagtt ggggaactgc ttaaccgtgg taactggctt
     3121 tcgcagagca cagcaaccaa atctgtcctt ccagtgtagc cggactttgg cgcacacttc
     3181 aagagcaacc gcgtgtttag ctaaacaaat cctctgcgaa ctcccagtta ccaatggctg
     3241 ctgccagtgg cgttttaccg tgcttttccg ggttggactc aagtgaacag ttaccggata
     3301 aggcgcagca gtcgggctga acggggagtt cttgcttaca gcccagcttg gagcgaacga
     3361 cctacaccga gccgagatac cagtgtgtga gctatgagaa agcgccacac ttcccgtaag
     3421 ggagaaaggc ggaacaggta tccggtaaac ggcagggtcg gaacaggaga gcgcaagagg
     3481 gagcgacccg ccggaaacgg tggggatctt taagtcctgt cgggtttcgc ccgtactgtc
     3541 agattcatgg ttgagcctca cggctcccac agatgcaccg gaaaagcgtc tgtttatgtg
     3601 aactctggca ggagggcgga gcctatggaa aaacgccacc ggcgcggccc tgctgttttg
     3661 cctcacatgt tagtcccctg cttatccacg gaatctgtgg gtaactttgt atgtgtccgc
     3721 agcgcccgcc gcagtctcac gcccggagcg tagcgaccga gtgacggcct atttggcttg
     3781 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     3841 tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     3901 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     3961 tgctaaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     4021 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     4081 tgctcatccg gagttccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     4141 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     4201 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     4261 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     4321 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     4381 ccccgttttc actatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     4441 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     4501 acaacagtac tgcgatgagt ggcagggcgg ggcgtaaacg ccatgggcat gtagtcaaaa
     4561 gcctccggtc ggaggctttt gacttggctg aggagtgccg ttaattaagt ttaaaccgcc
     4621 ggagatttgg tggtgtgcag gattttaacc tggatcatga tagacagttc agaacgacgt
     4681 ttttccgcac gtttcagggt tttttccagc caacggtttt cgttcagctg gttgtgagaa
     4741 gactggatga agattttacg gcagcactgg tagtgttcag aaaccataac gttcagacca
     4801 acaccgttgt cacgcaggtt ccacagaccg atctggttac gcgcgttttt ttcgtagtac
     4861 agtttgcacg cccagatttt cagggtgtga ccgttaccac gcagttcctg gttgtaccat
     4921 tccaggattt tttccgcgca gtccgcgcac ggagaccaag aagagtacca gttgatggtg
     4981 aactgacccg ggttgtcacg caggtattct tcaactttac ggatagagaa gatttccgcg
     5041 tggataccac gttcggtacc agactgcggt ttgttaaccg cgtaacccca gaagcacgca
     5101 cgacgttcac cacgacgttt cagttcgaac agaacgtagc aacggtgaga aacagatttt
     5161 ttgttgttga agaactgttt tttgaaggtg tagatgtcca gtttttcgtg gatacgaacg
     5221 tattccgcgt cggtcatctt cagattccgg tttacttaat ctcgacttat aacattttaa
     5281 ttttattttc tccattactg tcttgaatta ctaaagccca aggtttatat tctggagcat
     5341 ctgaagttaa tagcattaca ttttcatctg tactttcatc ataagcagta tgaactaaaa
     5401 tatcactttc tggtttattc ccaattactt cctctacttc ttctggtaac attagaattg
     5461 attcttgaat cactagttgt tttcctgttt ctttttcaat gatgtcagat aaatttgtca
     5521 tgttgaaatt cctcctaatt tttgtattat tagctcatcg cttcgagaac cacaccgtct
     5581 tgttccatct ggtcgagacg ggagagcaat ttgcgggtga tttgctcgag ctgatctttt
     5641 tctgttgtgc tgagcgcgga ccagagttga tgcaggcagt tatgctgcgg tggtaaaacc
     5701 tcgcgcaaaa actcgtgacc tttttccgtt aattgcagat gcaggcagcg gcgatcgtta
     5761 tcgctttcac gacgttcgat ccaaccgcgt ttttccagtt catcggcaat acgcgtcgcg
     5821 ttggtacggg atgatccaag agcacaactt aattcagaag gctgaatact gtggttttcc
     5881 tgagactcca gcgtaatcaa cgccataaac aacgtctcgt taattccctg agccttcagc
     5941 attttattgc ggttctccag cagcttgctt tgcatgtgca tgcaaagacg agtcagaagg
     6001 atctcctgat aaggaaaatc ttcgtggcgg ctggcgcgaa attttagcat ttgttcaatg
     6061 ggcgtaaacg aactatccat ttgggtatga cctcattaat ttcagccgtt agatagttcc
     6121 gcaaaccttc tcaatcaatt ccgccgggaa ttgcatcgac tgcatgatgt gttcgatcat
     6181 gctgcattta cggccggtgt tggtgttggt gatcacccaa tacggcgtgc ctggcacatg
     6241 tttcggcttg gtctgattac catttttcag cagcgtttgt tcatctgccg caaagtaaac
     6301 gcgtgtacga ccgtgcaacg attccgttgc ttcggcaaac gcctgggcgt caagagaata
     6361 tagtgtagac aacagcagca taaagcgatt gaccgctcgc ttttgctctg cgtattcatc
     6421 cgaaagcaga agttcacgca ttgcgcgaac cttgtcttta atcgttttga ccggcttcgc
     6481 ttcgacgata gcaggtgacg caacgcgaac ctctttcgtc accggagcag caggctgtga
     6541 tgcggcggaa aatttcaaca tacgccgtaa aatgtcggat gcgctctcgc cgatatgctt
     6601 agtgtggctg gcaatatagc tgtagagttc atcatcaact tcaatcgttt tcatcttaat
     6661 ccagtgcagt gtcttagctg aattattttt tcgcgggtga aacgactcct ggtttgtaca
     6721 aagccagcag ttcgtccacc tttttacgtg tgccgccgtt gctgcttata ctgcgtcgaa
     6781 ctttgacgac atgcaatttt gcgcgctgat accactcacg cgttaacatc gtatcgtgat
     6841 tggagatcag cactggaata tggcgctcaa ccagaccttc ggcgatctcc gccagatgcg
     6901 cttgttgttc aagcgtaaaa ctgtttgtgt gatacgccgt aaagttggcg gtcgcagaca
     6961 gcggtgcata aggcggatcg caatagacga cggatgcatc atctgcgcgc gccatgctat
     7021 cggcgtaaga ctcacaatag aaaaaggcat tctgcgcttt ttcagcgaag tgatacaact
     7081 ctgcttccgg gaaatagggt tttttgtagc ggccgaacgg cacgttaaac tcaccgcgca
     7141 gattgtaacg acacaggccg ttgtaaccgt agcggttcaa atataaaaac agtaccgccc
     7201 gacggaacgg atcctggctt ttgttgaact cttcgcggaa ctgatagtaa acctcggcgc
     7261 aatttgtttc gggaacaaac agctcgcgtg cggcctgtac gtactcatca gtacgcatct
     7321 tcacaatgtt atagagactg atcaggtcgc tattgatatc ggcaaggata taacgagaaa
     7381 agtcggtgtt gagaaacacc gacccggcac ctacaaaagg ctcaaccaga cattcgccct
     7441 tgggcaaatg ccgtttaata tcatcaagca ggggatactt gccccctgcc cacttcaaaa
     7501 aagcgcgatt tttcttcatg ggttatgtcc ggtttactga tattaaatta tgctcgccag
     7561 aggcaacttc cgcctttctt ctgcaccaga tcgagacggg cttcatgagc tgcaatctct
     7621 tcatctgtcg caaaaacaac gcgtaactta cttgcctgac gtacaatgcg ctgaattgtt
     7681 gcttcacctt gttgctgttg tgtctctcct tccatcgcaa aagccatcga cgtttgacca
     7741 ccggtcatcg ccagataaac ttccgcaagg atctgggcat cgagtaatgc cccgtgcagc
     7801 gttcgtttac tgttatctat ttcgtagcga gcacataacg catcgaggct gttgcgctta
     7861 ccgggaaaca ttttcctcgc caccgcaagg ctatcggtga ccttacagaa agtattggtc
     7921 ttcggaatat cgcgcttaag caacgaaaac tcgtagtcca taaagccgat atcgaacgct
     7981 gcgttatgga tcaccaactc cgcgccgcga atatagtcca tgaactcatc ggctacttcg
     8041 gcaaacgtgg gcttatcgag caaaaattca tcggcaatac catgtacgcc aaaggcttcc
     8101 ggatccacca gccgatcggg tttgagataa acatggaagt tattgcccgt caggcgacgg
     8161 ttcaccactt caacggcacc aatctcaatg atcttgtggc cttcatagtg cgcaccaatc
     8221 tggttcatac cggtggttgc ggtagcgaga acgatctggc gtgtaattgc agtgctcata
     8281 gcggtcattt atgtcagact tgagttgtac gcgtccaaaa aaacgggtat ggagaaacag
     8341 tagagagttg cgataaaaag cgtcaggtag gatccgctaa tcttatggat aaaaatgcta
     8401 tggcatagca aagtgtgacg ccgtgcaaat aatcaatgtg gacttttctg ccgtgattat
     8461 agacactttt gttacgcgtt tttgtcatgg ctttggtccc gctttgttac agaatgcttt
     8521 taataagcgg ggttaccggt tgggttagcg agaagagcca gtaaaagacg cagtgacggc
     8581 aatgtctgat gcaatatgga caattggttt cttctctgaa tggtgggagt atgaaaagt
//
