LOCUS       V015493                 7825 bp    DNA     circular SYN 07-DEC-2019
DEFINITION  Exported.
ACCESSION   V015493
VERSION     V015493
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 7825)
  AUTHORS   Alvarez B, Mencia M, de Lorenzo V, Fernandez LA.
  TITLE     In vivo mutagenesis of target DNA in E. coli chromosome with
            processive fusions of base deaminases and T7 RNA polymerase
            terminated by RNA-guided dCas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7825)
  AUTHORS   Alvarez B, Fernandez LA.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-SEP-2019) Microbial Biotechnology, CNB-CSIC (Centro
            Nacional de Biotecnologia-CSIC), Darwin 3, Cantoblanco, MADRID,
            Madrid 28049, Spain
REFERENCE   3  (bases 1 to 7825)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (13-SEP-2019) Microbial Biotechnology, CNB-CSIC (Centro Nacional de
            Biotecnologia-CSIC), Darwin 3, Cantoblanco, MADRID, Madrid 28049,
            Spain"
FEATURES             Location/Qualifiers
     source          1..7825
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5..99
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             231..1043
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     oriT            1213..1321
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(1385..2530)
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     rep_origin      complement(2923..3454)
                     /direction=LEFT
                     /label="oriV"
                     /note="incP origin of replication"
     terminator      complement(3585..3671)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(3717..4340)
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     promoter        4356..4411
                     /label="tetR/tetA promoters"
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             4448..4480
                     /label="T7 tag (gene 10 leader)"
                     /note="leader peptide from bacteriophage T7 gene 10"
     CDS             4481..5071
                     /label="AID"
                     /note="human activation-induced cytidine deaminase
                     (Dickerson et al., 2003)"
     misc_feature    5078..5161
                     /gene="AID-T7RNAP"
                     /label="Linker"
                     /note="Linker"
     CDS             5168..7816
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
ORIGIN
        1 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
       61 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggggtccc caataattac
      121 gatttaaatt tgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
      181 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
      241 ttcagcgtga aacgagctgt agccgtccgc gtctgaacag caacatggat gcggatctgt
      301 atggctataa atgggcgcgt gataacgtgg gtcagagcgg cgcgaccatt tatcgtctgt
      361 atggcaaacc ggatgcgccg gaactgtttc tgaaacatgg caaaggcagc gtggcgaacg
      421 atgtgaccga tgaaatggtg cgtctgaact ggctgaccga atttatgccg ctgccgacca
      481 ttaaacattt tattcgcacc ccggatgatg cgtggctgct gaccaccgcg attccgggca
      541 aaaccgcgtt tcaggtgctg gaagaatatc cggatagcgg cgaaaacatt gtggatgcgc
      601 tggccgtgtt tctgcgtcgt ctgcatagca ttccggtgtg caactgcccg tttaacagcg
      661 atcgtgtgtt tcgtctggcc caggcgcaga gccgtatgaa caacggcctg gtggatgcga
      721 gcgattttga tgatgaacgt aacggctggc cggtggaaca ggtgtggaaa gaaatgcata
      781 aactgctgcc gtttagcccg gatagcgtgg tgacccacgg cgattttagc ctggataacc
      841 tgattttcga tgaaggcaaa ctgattggct gcattgatgt gggccgtgtg ggcattgcgg
      901 atcgttatca ggatctggcc attctgtgga actgcctggg cgaatttagc ccgagcctgc
      961 aaaaacgtct gtttcagaaa tatggcattg ataatccgga tatgaacaaa ctgcaatttc
     1021 atctgatgct ggatgaattt ttctaataat taattggacc gcggtccgcg cgttgtcctt
     1081 ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt
     1141 tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca
     1201 acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt
     1261 cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc
     1321 gtaggccggc cgcgatgcag gtggctgctg aacccccagc cggaactgac cccacaaggc
     1381 cctagcgttt gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc
     1441 ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat
     1501 ccgatccgca catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc
     1561 tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga
     1621 atttcgtgta gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc
     1681 aacgggacgt tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt
     1741 ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc
     1801 attcctcggc cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct
     1861 cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct
     1921 gctgccacac cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca
     1981 cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt
     2041 tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg
     2101 gccacggcgc aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt
     2161 tcagcaacgc ggcctgcttg gcttcgctga cctgttttgc caggtcctcg ccggcggttt
     2221 ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg
     2281 ccgcctcctg ttcgagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag
     2341 ggggagccag ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg actatcgagc
     2401 cgacggactg gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg
     2461 catcctcggc ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg
     2521 tccgattcat tcaccctcct tgcgggattg ccccggaatt aattccccgg atcgatccgt
     2581 cgatcttgat cccctgcgcc atcagatcct tggcggcaag aaagccatcc agtttacttt
     2641 gcagggcttc ccaaccttac cagagggcgc cccagctggc aattccggtt cgcttgctgt
     2701 ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct
     2761 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     2821 gcaccgtttc tgcggactgg ctttctacgt ggctgccatt tttggggtga ggccgttcgc
     2881 ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc
     2941 gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt
     3001 taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc
     3061 cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa
     3121 atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg
     3181 cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca
     3241 ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg
     3301 aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg
     3361 cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca
     3421 agttttccgc gaggtatcca caacgccggc ggccctacat ggctctgctg tagtgagtgg
     3481 gttgcgctcc ggcagcggtc ctgatccccc gcagaaaaaa aggatctcaa gaagatcctt
     3541 tgatcttttc tacggcgcgc cgagctcgaa ttctagggcg gcggatttgt cctactcagg
     3601 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     3661 cgttttattt gatgcctcta gcacgcctgc caggaattgg ggatcggaga tctttaagac
     3721 ccactttcac atttaagttg tttttctaat ccgcatataa tcaattcaag gccgaataag
     3781 aaggctggct ctgcaccttg gtgatcaaat aattcgatag cttgtcgtaa taatggcggc
     3841 atactatcag tagtaggtgt ttccctttct tctttagcga cttgatgctc ttgatcttcc
     3901 aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg catataatgc attctctagt
     3961 gaaaaacctt gttggcataa aaaggctaat tgattttcga gagtttcata ctgtttttct
     4021 gtaggccgtg tacctaaatg tacttttgct ccatcgcgat gacttagtaa agcacatcta
     4081 agacttttag cgttattacg taaaaaatct tgccagcttt ccccttctaa agggcaaaag
     4141 tgagtatggt gcctatctaa catctcaatg gctaaggcgt cgagcaaagc ccgcttattt
     4201 tttacatgcc aatacaatgt aggctgctct acacctagct tctgggcgag tttacgggtt
     4261 gttaaacctt cgattccgac ctcattaagc agctctaatg cgctgttaat cactttactt
     4321 ttatctaatc tggacatcat taattcctaa tttttgttga cactctatca ttgatagagt
     4381 tattttacca ctccctatca gtgatagaga aaagtgaatc tagagtcgac aagaaggaga
     4441 tatacatatg gcaagcatga ccggtggtca gcagatgggt gatagcctgc tgatgaatcg
     4501 tcgtaaattt ctgtaccagt ttaaaaacgt tcgttgggca aaaggtcgtc gtgaaaccta
     4561 tctgtgttat gttgttaaac gtcgtgatag cgcaaccagc tttagcctgg attttggtta
     4621 tctgcgtaac aaaaatggtt gccatgttga actgctgttt ctgcgttata ttagcgattg
     4681 ggatttagat ccgggtcgtt gttatcgtgt tacctggttt accagttgga gcccgtgtta
     4741 tgactgtgcc cgtcatgttg cagattttct gcgtggtaat ccgaatctga gcctgcgtat
     4801 ttttaccgca cgtctgtatt tttgcgaaga tcgtaaagca gaaccggaag gtctgcgtcg
     4861 tctgcatcgt gccggtgttc agattgcaat tatgaccttc aaagactact tctactgctg
     4921 gaataccttt gtggaaaatc atgaacgtac ctttaaagca tgggaaggcc tgcatgaaaa
     4981 ttcagttcgt ctgagccgtc agctgcgtcg tattctgctg ccgctgtatg aagttgatga
     5041 tctgcgtgat gcatttcgta ccctgggtct gctgcagggt ggtggtagcg gtggtggttc
     5101 aggtggtggc agtggcggtg gaagtggtgg cggttctggc ggaggatcag gtggcggtag
     5161 caagcttatg aacacgatta acatcgctaa gaacgacttc tctgacatcg aactggctgc
     5221 tatcccgttc aacactctgg ctgaccatta cggtgagcgt ttagctcgcg aacagttggc
     5281 ccttgagcat gagtcttacg agatgggtga agcacgcttc cgcaagatgt ttgagcgtca
     5341 acttaaagct ggtgaggttg cggataacgc tgccgccaag cctctcatca ctaccctact
     5401 ccctaagatg attgcacgca tcaacgactg gtttgaggaa gtgaaagcta agcgcggcaa
     5461 gcgcccgaca gccttccagt tcctgcaaga aatcaagccg gaagccgtag cgtacatcac
     5521 cattaagacc actctggctt gcctaaccag tgctgacaat acaaccgttc aggctgtagc
     5581 aagcgcaatc ggtcgggcca ttgaggacga ggctcgcttc ggtcgtatcc gtgaccttga
     5641 agctaagcac ttcaagaaaa acgttgagga acaactcaac aagcgcgtag ggcacgtcta
     5701 caagaaagca tttatgcaag ttgtcgaggc tgacatgctc tctaagggtc tactcggtgg
     5761 cgaggcgtgg tcttcgtggc ataaggaaga ctctattcat gtaggagtac gctgcatcga
     5821 gatgctcatt gagtcaaccg gaatggttag cttacaccgc caaaatgctg gcgtagtagg
     5881 tcaagactct gagactatcg aactcgcacc tgaatacgct gaggctatcg caacccgtgc
     5941 aggtgcgctg gctggcatct ctccgatgtt ccaaccttgc gtagttcctc ctaagccgtg
     6001 gactggcatt actggtggtg gctattgggc taacggtcgt cgtcctctgg cgctggtgcg
     6061 tactcacagt aagaaagcac tgatgcgcta cgaagacgtt tacatgcctg aggtgtacaa
     6121 agcgattaac attgcgcaaa acaccgcatg gaaaatcaac aagaaagtcc tagcggtcgc
     6181 caacgtaatc accaagtgga agcattgtcc ggtcgaggac atccctgcga ttgagcgtga
     6241 agaactcccg atgaaaccgg aagacatcga catgaatcct gaggctctca ccgcgtggaa
     6301 acgtgctgcc gctgctgtgt accgcaagga caaggctcgc aagtctcgcc gtatcagcct
     6361 tgagttcatg cttgagcaag ccaataagtt tgctaaccat aaggccatct ggttccctta
     6421 caacatggac tggcgcggtc gtgtttacgc tgtgtcaatg ttcaacccgc aaggtaacga
     6481 tatgaccaaa ggactgctta cgctggcgaa aggtaaacca atcggtaagg aaggttacta
     6541 ctggctgaaa atccacggtg caaactgtgc gggtgtcgat aaggttccgt tccctgagcg
     6601 catcaagttc attgaggaaa accacgagaa catcatggct tgcgctaagt ctccactgga
     6661 gaacacttgg tgggctgagc aagattctcc gttctgcttc cttgcgttct gctttgagta
     6721 cgctggggta cagcaccacg gcctgagcta taactgctcc cttccgctgg cgtttgacgg
     6781 gtcttgctct ggcatccagc acttctccgc gatgctccga gatgaggtag gtggtcgcgc
     6841 ggttaacttg cttcctagtg aaaccgttca ggacatctac gggattgttg ctaagaaagt
     6901 caacgagatt ctacaagcag acgcaatcaa tgggaccgat aacgaagtag ttaccgtgac
     6961 cgatgagaac actggtgaaa tctctgagaa agtcaagctg ggcactaagg cactggctgg
     7021 tcaatggctg gcttacggtg ttactcgcag tgtgactaag cgttcagtca tgacgctggc
     7081 ttacgggtcc aaagagttcg gcttccgtca acaagtgctg gaagatacca ttcagccagc
     7141 tattgattcc ggcaagggtc tgatgttcac tcagccgaat caggctgctg gatacatggc
     7201 taagctgatt tgggaatctg tgagcgtgac ggtggtagct gcggttgaag caatgaactg
     7261 gcttaagtct gctgctaagc tgctggctgc tgaggtcaaa gataagaaga ctggagagat
     7321 tcttcgcaag cgttgcgctg tgcattgggt aactcctgat ggtttccctg tgtggcagga
     7381 atacaagaag cctattcaga cgcgcttgaa cctgatgttc ctcggtcagt tccgcttaca
     7441 gcctaccatt aacaccaaca aagatagcga gattgatgca cacaaacagg agtctggtat
     7501 cgctcctaac tttgtacaca gccaagacgg tagccacctt cgtaagactg tagtgtgggc
     7561 acacgagaag tacggaatcg aatcttttgc actgattcac gactccttcg gtaccattcc
     7621 ggctgacgct gcgaacctgt tcaaagcagt gcgcgaaact atggttgaca catatgagtc
     7681 ttgtgatgta ctggctgatt tctacgacca gttcgctgac cagttgcacg agtctcaatt
     7741 ggacaaaatg ccagcacttc cggctaaagg taacttgaac ctccgtgaca tcttagagtc
     7801 ggacttcgcg ttcgcgtaaa ctagt
//
