LOCUS       V015399                 8149 bp    DNA     circular SYN 06-OCT-2020
DEFINITION  Exported.
ACCESSION   V015399
VERSION     V015399
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8149)
  AUTHORS   Alvarez B, Mencia M, de Lorenzo V, Fernandez LA.
  TITLE     In vivo diversification of target genomic sites using processive T7
            RNA polymerase-base deaminase fusions blocked by RNA-guided dCas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8149)
  AUTHORS   Alvarez B, Fernandez LA.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-JUL-2020) Microbial Biotechnology, CNB-CSIC (Centro
            Nacional de Biotecnologia-CSIC), Darwin 3, Cantoblanco, Madrid,
            Madrid 28049, Spain
REFERENCE   3  (bases 1 to 8149)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (22-JUL-2020) Microbial Biotechnology, CNB-CSIC (Centro Nacional de
            Biotecnologia-CSIC), Darwin 3, Cantoblanco, Madrid, Madrid 28049,
            Spain"
FEATURES             Location/Qualifiers
     source          1..8149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(32..118)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(164..787)
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     promoter        803..858
                     /label="tetR/tetA promoters"
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             898..1221
                     /label="HP-thioredoxin"
                     /note="His-Patch thioredoxin"
     CDS             1237..1251
                     /label="enterokinase site"
                     /note="enterokinase recognition and cleavage site"
     CDS             1252..1842
                     /label="AID"
                     /note="human activation-induced cytidine deaminase
                     (Dickerson et al., 2003)"
     misc_feature    1849..1932
                     /gene="TrxA-AID-T7RNAP"
                     /label="Linker"
                     /note="Linker"
     CDS             1939..4587
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     terminator      4601..4695
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             4827..5639
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     oriT            5809..5917
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(5981..7126)
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     rep_origin      complement(7519..8050)
                     /direction=LEFT
                     /label="oriV"
                     /note="incP origin of replication"
ORIGIN
        1 ggcgcgccga gctcgaattc tagggcggcg gatttgtcct actcaggaga gcgttcaccg
       61 acaaacaaca gataaaacga aaggcccagt ctttcgactg agcctttcgt tttatttgat
      121 gcctctagca cgcctgccag gaattgggga tcggagatct ttaagaccca ctttcacatt
      181 taagttgttt ttctaatccg catataatca attcaaggcc gaataagaag gctggctctg
      241 caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata ctatcagtag
      301 taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat acgcaaccta
      361 aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa aaaccttgtt
      421 ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta ggccgtgtac
      481 ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaga cttttagcgt
      541 tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga gtatggtgcc
      601 tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt acatgccaat
      661 acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt aaaccttcga
      721 ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta tctaatctgg
      781 acatcattaa ttcctaattt ttgttgacac tctatcattg atagagttat tttaccactc
      841 cctatcagtg atagagaaaa gtgaatctag agtcgacaag aaggagatat acatatgtct
      901 gataaaatta ttcatctgac tgatgattct tttgatactg atgtacttaa ggcagatggt
      961 gcaatcctgg ttgatttctg ggcacactgg tgcggtccgt gcaaaatgat cgctccgatt
     1021 ctggatgaaa tcgctgacga atatcagggc aaactgaccg ttgcaaaact gaacatcgat
     1081 cacaacccgg gcactgcgcc gaaatatggc atccgtggta tcccgactct gctgctgttc
     1141 aaaaacggtg aagtggcggc aaccaaagtg ggtgcactgt ctaaaggtca gttgaaagag
     1201 ttcctcgacg ctaacctggc cggctctgga tccggtgatg acgatgacaa ggatagcctg
     1261 ctgatgaatc gtcgtaaatt tctgtaccag tttaaaaacg ttcgttgggc aaaaggtcgt
     1321 cgtgaaacct atctgtgtta tgttgttaaa cgtcgtgata gcgcaaccag ctttagcctg
     1381 gattttggtt atctgcgtaa caaaaatggt tgccatgttg aactgctgtt tctgcgttat
     1441 attagcgatt gggatttaga tccgggtcgt tgttatcgtg ttacctggtt taccagttgg
     1501 agcccgtgtt atgactgtgc ccgtcatgtt gcagattttc tgcgtggtaa tccgaatctg
     1561 agcctgcgta tttttaccgc acgtctgtat ttttgcgaag atcgtaaagc agaaccggaa
     1621 ggtctgcgtc gtctgcatcg tgccggtgtt cagattgcaa ttatgacctt caaagactac
     1681 ttctactgct ggaatacctt tgtggaaaat catgaacgta cctttaaagc atgggaaggc
     1741 ctgcatgaaa attcagttcg tctgagccgt cagctgcgtc gtattctgct gccgctgtat
     1801 gaagttgatg atctgcgtga tgcatttcgt accctgggtc tgctgcaggg tggtggtagc
     1861 ggtggtggtt caggtggtgg cagtggcggt ggaagtggtg gcggttctgg cggaggatca
     1921 ggtggcggta gcaagcttat gaacacgatt aacatcgcta agaacgactt ctctgacatc
     1981 gaactggctg ctatcccgtt caacactctg gctgaccatt acggtgagcg tttagctcgc
     2041 gaacagttgg cccttgagca tgagtcttac gagatgggtg aagcacgctt ccgcaagatg
     2101 tttgagcgtc aacttaaagc tggtgaggtt gcggataacg ctgccgccaa gcctctcatc
     2161 actaccctac tccctaagat gattgcacgc atcaacgact ggtttgagga agtgaaagct
     2221 aagcgcggca agcgcccgac agccttccag ttcctgcaag aaatcaagcc ggaagccgta
     2281 gcgtacatca ccattaagac cactctggct tgcctaacca gtgctgacaa tacaaccgtt
     2341 caggctgtag caagcgcaat cggtcgggcc attgaggacg aggctcgctt cggtcgtatc
     2401 cgtgaccttg aagctaagca cttcaagaaa aacgttgagg aacaactcaa caagcgcgta
     2461 gggcacgtct acaagaaagc atttatgcaa gttgtcgagg ctgacatgct ctctaagggt
     2521 ctactcggtg gcgaggcgtg gtcttcgtgg cataaggaag actctattca tgtaggagta
     2581 cgctgcatcg agatgctcat tgagtcaacc ggaatggtta gcttacaccg ccaaaatgct
     2641 ggcgtagtag gtcaagactc tgagactatc gaactcgcac ctgaatacgc tgaggctatc
     2701 gcaacccgtg caggtgcgct ggctggcatc tctccgatgt tccaaccttg cgtagttcct
     2761 cctaagccgt ggactggcat tactggtggt ggctattggg ctaacggtcg tcgtcctctg
     2821 gcgctggtgc gtactcacag taagaaagca ctgatgcgct acgaagacgt ttacatgcct
     2881 gaggtgtaca aagcgattaa cattgcgcaa aacaccgcat ggaaaatcaa caagaaagtc
     2941 ctagcggtcg ccaacgtaat caccaagtgg aagcattgtc cggtcgagga catccctgcg
     3001 attgagcgtg aagaactccc gatgaaaccg gaagacatcg acatgaatcc tgaggctctc
     3061 accgcgtgga aacgtgctgc cgctgctgtg taccgcaagg acaaggctcg caagtctcgc
     3121 cgtatcagcc ttgagttcat gcttgagcaa gccaataagt ttgctaacca taaggccatc
     3181 tggttccctt acaacatgga ctggcgcggt cgtgtttacg ctgtgtcaat gttcaacccg
     3241 caaggtaacg atatgaccaa aggactgctt acgctggcga aaggtaaacc aatcggtaag
     3301 gaaggttact actggctgaa aatccacggt gcaaactgtg cgggtgtcga taaggttccg
     3361 ttccctgagc gcatcaagtt cattgaggaa aaccacgaga acatcatggc ttgcgctaag
     3421 tctccactgg agaacacttg gtgggctgag caagattctc cgttctgctt ccttgcgttc
     3481 tgctttgagt acgctggggt acagcaccac ggcctgagct ataactgctc ccttccgctg
     3541 gcgtttgacg ggtcttgctc tggcatccag cacttctccg cgatgctccg agatgaggta
     3601 ggtggtcgcg cggttaactt gcttcctagt gaaaccgttc aggacatcta cgggattgtt
     3661 gctaagaaag tcaacgagat tctacaagca gacgcaatca atgggaccga taacgaagta
     3721 gttaccgtga ccgatgagaa cactggtgaa atctctgaga aagtcaagct gggcactaag
     3781 gcactggctg gtcaatggct ggcttacggt gttactcgca gtgtgactaa gcgttcagtc
     3841 atgacgctgg cttacgggtc caaagagttc ggcttccgtc aacaagtgct ggaagatacc
     3901 attcagccag ctattgattc cggcaagggt ctgatgttca ctcagccgaa tcaggctgct
     3961 ggatacatgg ctaagctgat ttgggaatct gtgagcgtga cggtggtagc tgcggttgaa
     4021 gcaatgaact ggcttaagtc tgctgctaag ctgctggctg ctgaggtcaa agataagaag
     4081 actggagaga ttcttcgcaa gcgttgcgct gtgcattggg taactcctga tggtttccct
     4141 gtgtggcagg aatacaagaa gcctattcag acgcgcttga acctgatgtt cctcggtcag
     4201 ttccgcttac agcctaccat taacaccaac aaagatagcg agattgatgc acacaaacag
     4261 gagtctggta tcgctcctaa ctttgtacac agccaagacg gtagccacct tcgtaagact
     4321 gtagtgtggg cacacgagaa gtacggaatc gaatcttttg cactgattca cgactccttc
     4381 ggtaccattc cggctgacgc tgcgaacctg ttcaaagcag tgcgcgaaac tatggttgac
     4441 acatatgagt cttgtgatgt actggctgat ttctacgacc agttcgctga ccagttgcac
     4501 gagtctcaat tggacaaaat gccagcactt ccggctaaag gtaacttgaa cctccgtgac
     4561 atcttagagt cggacttcgc gttcgcgtaa actagtcttg gactcctgtt gatagatcca
     4621 gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg ccgggcgttt
     4681 tttattggtg agaatccagg ggtccccaat aattacgatt taaatttgtg tctcaaaatc
     4741 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     4801 acataaacag taatacaagg ggtgttatga gccatattca gcgtgaaacg agctgtagcc
     4861 gtccgcgtct gaacagcaac atggatgcgg atctgtatgg ctataaatgg gcgcgtgata
     4921 acgtgggtca gagcggcgcg accatttatc gtctgtatgg caaaccggat gcgccggaac
     4981 tgtttctgaa acatggcaaa ggcagcgtgg cgaacgatgt gaccgatgaa atggtgcgtc
     5041 tgaactggct gaccgaattt atgccgctgc cgaccattaa acattttatt cgcaccccgg
     5101 atgatgcgtg gctgctgacc accgcgattc cgggcaaaac cgcgtttcag gtgctggaag
     5161 aatatccgga tagcggcgaa aacattgtgg atgcgctggc cgtgtttctg cgtcgtctgc
     5221 atagcattcc ggtgtgcaac tgcccgttta acagcgatcg tgtgtttcgt ctggcccagg
     5281 cgcagagccg tatgaacaac ggcctggtgg atgcgagcga ttttgatgat gaacgtaacg
     5341 gctggccggt ggaacaggtg tggaaagaaa tgcataaact gctgccgttt agcccggata
     5401 gcgtggtgac ccacggcgat tttagcctgg ataacctgat tttcgatgaa ggcaaactga
     5461 ttggctgcat tgatgtgggc cgtgtgggca ttgcggatcg ttatcaggat ctggccattc
     5521 tgtggaactg cctgggcgaa tttagcccga gcctgcaaaa acgtctgttt cagaaatatg
     5581 gcattgataa tccggatatg aacaaactgc aatttcatct gatgctggat gaatttttct
     5641 aataattaat tggaccgcgg tccgcgcgtt gtccttttcc gctgcataac cctgcttcgg
     5701 ggtcattata gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg
     5761 ccaaagggtt cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg
     5821 tgaagtaggc ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc
     5881 ggtgctcaac gggaatcctg ctctgcgagg ctggccgtag gccggccgcg atgcaggtgg
     5941 ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt
     6001 catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc
     6061 cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg
     6121 tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg
     6181 ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa
     6241 acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt
     6301 ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg
     6361 tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc
     6421 ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct
     6481 cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt
     6541 cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga
     6601 ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc
     6661 gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg
     6721 aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggctt
     6781 cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc
     6841 ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttcg agacgacgcg
     6901 aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc
     6961 gctcgatctt ggccgtagct tgctggacta tcgagccgac ggactggaag gtttcgcggg
     7021 gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt
     7081 cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg
     7141 ggattgcccc ggaattaatt ccccggatcg atccgtcgat cttgatcccc tgcgccatca
     7201 gatccttggc ggcaagaaag ccatccagtt tactttgcag ggcttcccaa ccttaccaga
     7261 gggcgcccca gctggcaatt ccggttcgct tgctgtccat aaaaccgccc agtctagcta
     7321 tcgccatgta agcccactgc aagctacctg ctttctcttt gcgcttgcgt tttcccttgt
     7381 ccagatagcc cagtagctga cattcatccg gggtcagcac cgtttctgcg gactggcttt
     7441 ctacgtggct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg
     7501 ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg
     7561 gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg
     7621 gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc
     7681 aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc
     7741 tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc
     7801 ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga
     7861 aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg
     7921 gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag
     7981 tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac
     8041 gccggcggcc ctacatggct ctgctgtagt gagtgggttg cgctccggca gcggtcctga
     8101 tcccccgcag aaaaaaagga tctcaagaag atcctttgat cttttctac
//
