LOCUS       sequence-273050-        5188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P3a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5188)
  AUTHORS   Richter MF, Zhao KT, Eton E, Lapinaite A, Newby GA, Thuronyi BW, 
            Wilson C, Koblan LW, Zeng J, Bauer DE, Doudna JA, Liu DR
  TITLE     Phage-assisted evolution of an adenine base editor with improved Cas
            domain compatibility and activity.
  JOURNAL   Nat Biotechnol. 2020 Jul;38(7):883-891. doi: 
            10.1038/s41587-020-0453-z. Epub 2020 Mar 16.
  PUBMED    32433547
REFERENCE   2  (bases 1 to 5188)
  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"; doi: 
            "10.1038/s41587-020-0453-z"; journalName: "Nat Biotechnol"; date: 
            "2020-07"; volume: "38"; issue: "7"; pages: "883-891"
COMMENT     Sequence Label: pMR-A-3
FEATURES             Location/Qualifiers
     source          1..5188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     301..320
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     554..571
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    586..726
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(652..671)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     812..834
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(828..1019)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4109..4900)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(4901..4972)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5000..5188,1..400)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
       61 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      121 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
      181 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
      241 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
      301 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      361 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      421 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
      481 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
      541 aacgaccgag cgcagcgagt cagtgagcga ggaagcggat gagcgcctga tgcggtattt
      601 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
      661 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
      721 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
      781 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
      841 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
      901 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
      961 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     1021 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     1081 aggatgctca cgatacgggt tactgatgat gaacaagagg acatccggtc aaataaaacg
     1141 aaaggctcag tcgaaagact gggcctttcg ttttgctgag gagacttagg gactctcaca
     1201 gctaacacca cgtcgtccct atctgctgcc ctaggtctat gagtggttgc tggataactt
     1261 tacgggcatg cataaggctc gtaggctata ttcagggaga ccacaacggt ttccctctac
     1321 aaataatttt gtttaacttt tactagagtg ggaccctacc tgcaggtgca gtaaggagga
     1381 aaaaaaaatg aacacgatta acatcgctaa gaacgacttc tctgacatcg aactggctgc
     1441 tatcccgttc aacactctgg ctgaccatta cggtgagcgt ttagctcgcg aacagttggc
     1501 ccttgagcat gagtcttacg agatgggtga agcacgcttc cgcaagatgt ttgagtgata
     1561 acttaaagct ggtgaggttg cggataacgc tgccgccaag cctctcatca ctaccctact
     1621 ccctaagatg attgcacgca tcaacgactg gtttgaggaa gtgaaagcta agcgcggcaa
     1681 gcgcccgaca gccttccagt tcctgcaaga aatcaagccg gaagccgtag cgtacatcac
     1741 cattaagacc actctggctt gcctaaccag tgctgacaat acaaccgttc aggctgtagc
     1801 aagcgcaatc ggtcgggcca ttgaggacga ggctcgcttc ggtcgtatcc gtgaccttaa
     1861 agctaagcac ttcaagaaaa acgttgagga acaactcaac aagcgcgtag ggcacgtcta
     1921 caagaaagca tttatgcaag ttgtcgaggc tgacatgctc tctaagggtc tactcggtgg
     1981 cgaggcgtgg tcttcgtggc ataaggaaga ctctattcat gtaggagtac gctgcatcga
     2041 gatgctcatt gagtcaaccg gaatggttag cttacaccgc caaaatgctg gcgtagtagg
     2101 tcaagactct gagactatcg aactcgcacc tgaatacgct gaggctatcg caacccgtgc
     2161 aggtgcgctg gctggcatct ctccgatgtt ccaaccttgc gtagttcctc ctaagccgtg
     2221 gactggcatt actggtggtg gctattgggc taacggtcgt cgtcctctgg cgctggtgcg
     2281 tactcacagt aagaaagcac tgatgcgcta cgaagacgtt tacatgcctg aggtgtacaa
     2341 agcgattaac attgcgcaaa acaccgcatg gaaaatcaac aagaaagtcc tagcggtcgc
     2401 caacgtaatc accaagtgga agcattgtcc ggtcgaggac atccctgcga ttgagcgtga
     2461 agaactcccg atgaaaccgg aagacatcga catgaatcct gaggctctca ccgcgtggaa
     2521 acgtgctgcc gctgctgtgt accgcaagga caaggctcgc aagtctcgcc gtatcagcct
     2581 tgagttcatg cttgagcaag ccaataagtt tgctaaccat aaggccatct ggttccctta
     2641 caacatggac tggcgcggtc gtgtttacgc tgtgtcaatg ttcaacccgc aaggtaacga
     2701 tatgaccaaa ggactgctta cgctggcgaa aggtaaacca atcggtaagg aaggttacta
     2761 ctggctgaaa atccacggtg caaactgtgc gggtgtcgat aaggttccgt tccctgagcg
     2821 catcaagttc attgaggaaa accacgagaa catcatggct tgcgctaagt ctccactgga
     2881 gaacacttgg tgggctgagc aagattctcc gttctgcttc cttgcgttct gctttgagta
     2941 cgctggggta cagcaccacg gcctgagcta taactgctcc cttccgctgg cgtttgacgg
     3001 gtcttgctct ggcatccagc acttctccgc gatgctccga gatgaggtag gtggtcgcgc
     3061 ggttaacttg cttcctagtg aaaccgttca ggacatctac gggattgttg ctaagaaagt
     3121 caacgagatt ctacaagcag acgcaatcaa tgggaccgat aacgaagtag ttaccgtgac
     3181 cgatgagaac actggtgaaa tctctgagaa agtcaagctg ggcactaagg cactggctgg
     3241 tcaatggctg gcttacggtg ttactcgcag tgtgactaag cgttcagtca tgacgctggc
     3301 ttacgggtcc aaagagttcg gcttccgtca acaagtgctg gaagatacca ttcagccagc
     3361 tattgattcc ggcaagggtc tgatgttcac tcagccgaat caggctgctg gatacatggc
     3421 taagctgatt tgggaatctg tgagcgtgac ggtggtagct gcggttgaag caatgaactg
     3481 gcttaagtct gctgctaagc tgctggctgc tgaggtcaaa gataagaaga ctggagagat
     3541 tcttcgcaag cgttgcgctg tgcattgggt aactcctgat ggtttccctg tgtggcagga
     3601 atacaagaag cctattcaga cgcgcttgaa cctgatgttc ctcggtcagt tccgcttaca
     3661 gcctaccatt aacaccaaca aagatagcga gattgatgca cacaaacagg agtctggtat
     3721 cgctcctaac tttgtacaca gccaagacgg tagccacctt cgtaagactg tagtgtgggc
     3781 acacgagaag tacggtatcg aatcttttgc actgattcac gactccttcg gtaccattcc
     3841 ggctgacgct gcgaacctgt tcaaagcagt gcgcgaaact atggttgaca catatgagtc
     3901 ttgtgatgta ctggctgatt tctacgacca gttcgctgac cagttgcacg agtctcaatt
     3961 ggacaaaatg ccagcacttc cggctaaagg taacttgaac ctccgtgaca tcttagagtc
     4021 cgacttcgcg ttcgcctaat ggcactcctc agcaaatata atgaccctct tgataaccca
     4081 agagggcatt ttttaatgcc catggcgttt atttgccgac taccttggtg atctcgcctt
     4141 tcacgtagtg gacaaattct tccaactgat ctgcgcgcga ggccaagcga tcttcttctt
     4201 gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc ggcaggcgct
     4261 ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact gcgctgtacc
     4321 aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg ggcggcgagt
     4381 tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga accggatcaa
     4441 agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct tttgtcagca
     4501 agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga atgtcattgc
     4561 gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga atgatgtcgt
     4621 cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca ggggaagccg
     4681 aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc cttacggtca
     4741 ccgtaaccag caaatcaata tcactgtgtg gcttcaggcc gccatccact gcggagccgt
     4801 acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca actacctctg
     4861 atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt tttcaatatt
     4921 attgaagcat ttatcagggt tattgtctca tgagcgtata catatttgaa tgtatttaga
     4981 aaaataggcc aaatagggtt tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5041 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccacctcttt
     5101 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     5161 cgtagttagg ccaccacttc aagaactc
//
