LOCUS       V015579                 8899 bp    DNA     circular SYN 06-OCT-2020
DEFINITION  Exported.
ACCESSION   V015579
VERSION     V015579
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8899)
  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  JOURNAL   Unpublished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 8899)
  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 8899)
  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            "Unpublished"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..8899
                     /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             895..1995
                     /label="MBP"
                     /note="maltose binding protein from E. coli"
     CDS             2002..2592
                     /label="AID"
                     /note="human activation-induced cytidine deaminase
                     (Dickerson et al., 2003)"
     misc_feature    2599..2682
                     /gene="MBPc-AID-T7RNAP"
                     /label="Linker"
                     /note="Linker"
     CDS             2689..5337
                     /note="T7 RNA polymerase from Escherichia phage T7.
                     Accession#: P00573"
                     /label="T7 RNA polymerase"
     terminator      5351..5445
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             5577..6389
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     oriT            6559..6667
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(6731..7876)
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     rep_origin      complement(8269..8800)
                     /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 acatatgaaa
      901 actgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct
      961 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat
     1021 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc
     1081 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg
     1141 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc
     1201 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg
     1261 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg
     1321 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt
     1381 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg
     1441 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac
     1501 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa
     1561 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat
     1621 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg
     1681 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa
     1741 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc
     1801 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccactatg
     1861 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat
     1921 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg
     1981 aaagacgcgc agactggatc cgatagcctg ctgatgaatc gtcgtaaatt tctgtaccag
     2041 tttaaaaacg ttcgttgggc aaaaggtcgt cgtgaaacct atctgtgtta tgttgttaaa
     2101 cgtcgtgata gcgcaaccag ctttagcctg gattttggtt atctgcgtaa caaaaatggt
     2161 tgccatgttg aactgctgtt tctgcgttat attagcgatt gggatttaga tccgggtcgt
     2221 tgttatcgtg ttacctggtt taccagttgg agcccgtgtt atgactgtgc ccgtcatgtt
     2281 gcagattttc tgcgtggtaa tccgaatctg agcctgcgta tttttaccgc acgtctgtat
     2341 ttttgcgaag atcgtaaagc agaaccggaa ggtctgcgtc gtctgcatcg tgccggtgtt
     2401 cagattgcaa ttatgacctt caaagactac ttctactgct ggaatacctt tgtggaaaat
     2461 catgaacgta cctttaaagc atgggaaggc ctgcatgaaa attcagttcg tctgagccgt
     2521 cagctgcgtc gtattctgct gccgctgtat gaagttgatg atctgcgtga tgcatttcgt
     2581 accctgggtc tgctgcaggg tggtggtagc ggtggtggtt caggtggtgg cagtggcggt
     2641 ggaagtggtg gcggttctgg cggaggatca ggtggcggta gcaagcttat gaacacgatt
     2701 aacatcgcta agaacgactt ctctgacatc gaactggctg ctatcccgtt caacactctg
     2761 gctgaccatt acggtgagcg tttagctcgc gaacagttgg cccttgagca tgagtcttac
     2821 gagatgggtg aagcacgctt ccgcaagatg tttgagcgtc aacttaaagc tggtgaggtt
     2881 gcggataacg ctgccgccaa gcctctcatc actaccctac tccctaagat gattgcacgc
     2941 atcaacgact ggtttgagga agtgaaagct aagcgcggca agcgcccgac agccttccag
     3001 ttcctgcaag aaatcaagcc ggaagccgta gcgtacatca ccattaagac cactctggct
     3061 tgcctaacca gtgctgacaa tacaaccgtt caggctgtag caagcgcaat cggtcgggcc
     3121 attgaggacg aggctcgctt cggtcgtatc cgtgaccttg aagctaagca cttcaagaaa
     3181 aacgttgagg aacaactcaa caagcgcgta gggcacgtct acaagaaagc atttatgcaa
     3241 gttgtcgagg ctgacatgct ctctaagggt ctactcggtg gcgaggcgtg gtcttcgtgg
     3301 cataaggaag actctattca tgtaggagta cgctgcatcg agatgctcat tgagtcaacc
     3361 ggaatggtta gcttacaccg ccaaaatgct ggcgtagtag gtcaagactc tgagactatc
     3421 gaactcgcac ctgaatacgc tgaggctatc gcaacccgtg caggtgcgct ggctggcatc
     3481 tctccgatgt tccaaccttg cgtagttcct cctaagccgt ggactggcat tactggtggt
     3541 ggctattggg ctaacggtcg tcgtcctctg gcgctggtgc gtactcacag taagaaagca
     3601 ctgatgcgct acgaagacgt ttacatgcct gaggtgtaca aagcgattaa cattgcgcaa
     3661 aacaccgcat ggaaaatcaa caagaaagtc ctagcggtcg ccaacgtaat caccaagtgg
     3721 aagcattgtc cggtcgagga catccctgcg attgagcgtg aagaactccc gatgaaaccg
     3781 gaagacatcg acatgaatcc tgaggctctc accgcgtgga aacgtgctgc cgctgctgtg
     3841 taccgcaagg acaaggctcg caagtctcgc cgtatcagcc ttgagttcat gcttgagcaa
     3901 gccaataagt ttgctaacca taaggccatc tggttccctt acaacatgga ctggcgcggt
     3961 cgtgtttacg ctgtgtcaat gttcaacccg caaggtaacg atatgaccaa aggactgctt
     4021 acgctggcga aaggtaaacc aatcggtaag gaaggttact actggctgaa aatccacggt
     4081 gcaaactgtg cgggtgtcga taaggttccg ttccctgagc gcatcaagtt cattgaggaa
     4141 aaccacgaga acatcatggc ttgcgctaag tctccactgg agaacacttg gtgggctgag
     4201 caagattctc cgttctgctt ccttgcgttc tgctttgagt acgctggggt acagcaccac
     4261 ggcctgagct ataactgctc ccttccgctg gcgtttgacg ggtcttgctc tggcatccag
     4321 cacttctccg cgatgctccg agatgaggta ggtggtcgcg cggttaactt gcttcctagt
     4381 gaaaccgttc aggacatcta cgggattgtt gctaagaaag tcaacgagat tctacaagca
     4441 gacgcaatca atgggaccga taacgaagta gttaccgtga ccgatgagaa cactggtgaa
     4501 atctctgaga aagtcaagct gggcactaag gcactggctg gtcaatggct ggcttacggt
     4561 gttactcgca gtgtgactaa gcgttcagtc atgacgctgg cttacgggtc caaagagttc
     4621 ggcttccgtc aacaagtgct ggaagatacc attcagccag ctattgattc cggcaagggt
     4681 ctgatgttca ctcagccgaa tcaggctgct ggatacatgg ctaagctgat ttgggaatct
     4741 gtgagcgtga cggtggtagc tgcggttgaa gcaatgaact ggcttaagtc tgctgctaag
     4801 ctgctggctg ctgaggtcaa agataagaag actggagaga ttcttcgcaa gcgttgcgct
     4861 gtgcattggg taactcctga tggtttccct gtgtggcagg aatacaagaa gcctattcag
     4921 acgcgcttga acctgatgtt cctcggtcag ttccgcttac agcctaccat taacaccaac
     4981 aaagatagcg agattgatgc acacaaacag gagtctggta tcgctcctaa ctttgtacac
     5041 agccaagacg gtagccacct tcgtaagact gtagtgtggg cacacgagaa gtacggaatc
     5101 gaatcttttg cactgattca cgactccttc ggtaccattc cggctgacgc tgcgaacctg
     5161 ttcaaagcag tgcgcgaaac tatggttgac acatatgagt cttgtgatgt actggctgat
     5221 ttctacgacc agttcgctga ccagttgcac gagtctcaat tggacaaaat gccagcactt
     5281 ccggctaaag gtaacttgaa cctccgtgac atcttagagt cggacttcgc gttcgcgtaa
     5341 actagtcttg gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg
     5401 ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccagg ggtccccaat
     5461 aattacgatt taaatttgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     5521 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     5581 gccatattca gcgtgaaacg agctgtagcc gtccgcgtct gaacagcaac atggatgcgg
     5641 atctgtatgg ctataaatgg gcgcgtgata acgtgggtca gagcggcgcg accatttatc
     5701 gtctgtatgg caaaccggat gcgccggaac tgtttctgaa acatggcaaa ggcagcgtgg
     5761 cgaacgatgt gaccgatgaa atggtgcgtc tgaactggct gaccgaattt atgccgctgc
     5821 cgaccattaa acattttatt cgcaccccgg atgatgcgtg gctgctgacc accgcgattc
     5881 cgggcaaaac cgcgtttcag gtgctggaag aatatccgga tagcggcgaa aacattgtgg
     5941 atgcgctggc cgtgtttctg cgtcgtctgc atagcattcc ggtgtgcaac tgcccgttta
     6001 acagcgatcg tgtgtttcgt ctggcccagg cgcagagccg tatgaacaac ggcctggtgg
     6061 atgcgagcga ttttgatgat gaacgtaacg gctggccggt ggaacaggtg tggaaagaaa
     6121 tgcataaact gctgccgttt agcccggata gcgtggtgac ccacggcgat tttagcctgg
     6181 ataacctgat tttcgatgaa ggcaaactga ttggctgcat tgatgtgggc cgtgtgggca
     6241 ttgcggatcg ttatcaggat ctggccattc tgtggaactg cctgggcgaa tttagcccga
     6301 gcctgcaaaa acgtctgttt cagaaatatg gcattgataa tccggatatg aacaaactgc
     6361 aatttcatct gatgctggat gaatttttct aataattaat tggaccgcgg tccgcgcgtt
     6421 gtccttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc
     6481 atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg
     6541 tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc
     6601 cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg
     6661 ctggccgtag gccggccgcg atgcaggtgg ctgctgaacc cccagccgga actgacccca
     6721 caaggcccta gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc
     6781 cgctgcctcg caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg
     6841 tggaatccga tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt
     6901 gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc
     6961 atatgaattt cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga
     7021 cctggcaacg ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca
     7081 ccgattccag gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg
     7141 acaggcattc ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc
     7201 aaagctcgta gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca
     7261 acacctgctg ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga
     7321 tcttcacgtc cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt
     7381 tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg
     7441 tgtccggcca cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg
     7501 tgtgtttcag caacgcggcc tgcttggctt cgctgacctg ttttgccagg tcctcgccgg
     7561 cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca
     7621 aacctgccgc ctcctgttcg agacgacgcg aacgctccac ggcggccgat ggcgcgggca
     7681 gggcaggggg agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacta
     7741 tcgagccgac ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg
     7801 tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt
     7861 caaacgtccg attcattcac cctccttgcg ggattgcccc ggaattaatt ccccggatcg
     7921 atccgtcgat cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt
     7981 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct
     8041 tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg
     8101 ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     8161 gggtcagcac cgtttctgcg gactggcttt ctacgtggct gccatttttg gggtgaggcc
     8221 gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag cgggccggga
     8281 gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc
     8341 cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag acaggttagc
     8401 ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct gtggacagcc
     8461 cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca agtgtcaagg
     8521 atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc agggcactta
     8581 tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg ttttcgccga
     8641 tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca
     8701 acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga
     8761 gggccaagtt ttccgcgagg tatccacaac gccggcggcc ctacatggct ctgctgtagt
     8821 gagtgggttg cgctccggca gcggtcctga tcccccgcag aaaaaaagga tctcaagaag
     8881 atcctttgat cttttctac
//
