LOCUS       sequence-273045-        5188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P3f.
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: pBT138b-proB
FEATURES             Location/Qualifiers
     source          1..5188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(66..857)
                     /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(858..929)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      957..1545
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1446..1465
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1699..1716
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1731..1871
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1797..1816)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1957..1979
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1973..2164)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 actcctcagc aaatataatg accctcttga taacccaaga gggcattttt taatgcccat
       61 ggcgtttatt tgccgactac cttggtgatc tcgcctttca cgtagtggac aaattcttcc
      121 aactgatctg cgcgcgaggc caagcgatct tcttcttgtc caagataagc ctgtctagct
      181 tcaagtatga cgggctgata ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca
      241 tccttcggcg cgattttgcc ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact
      301 acatttcgct catcgccagc ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt
      361 agcgcctcaa atagatcctg ttcaggaacc ggatcaaaga gttcctccgc cgctggacct
      421 accaaggcaa cgctatgttc tcttgctttt gtcagcaaga tagccagatc aatgtcgatc
      481 gtggctggct cgaagatacc tgcaagaatg tcattgcgct gccattctcc aaattgcagt
      541 tcgcgcttag ctggataacg ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct
      601 acagcgcgga gaatctcgct ctctccaggg gaagccgaag tttccaaaag gtcgttgatc
      661 aaagctcgcc gcgttgtttc atcaagcctt acggtcaccg taaccagcaa atcaatatca
      721 ctgtgtggct tcaggccgcc atccactgcg gagccgtaca aatgtacggc cagcaacgtc
      781 ggttcgagat ggcgctcgat gacgccaact acctctgata gttgagtcga tacttcggcg
      841 atcaccgctt ccctcatact cttccttttt caatattatt gaagcattta tcagggttat
      901 tgtctcatga gcgtatacat atttgaatgt atttagaaaa ataggccaaa tagggtttga
      961 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1021 gtggtttgtt tgccggatca agagctacca cctctttttc cgaaggtaac tggcttcagc
     1081 agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag
     1141 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1201 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1261 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1321 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1381 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1441 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1501 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1561 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1621 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1681 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggatgagcg cctgatgcgg
     1741 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     1801 atctgctctg atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg
     1861 tcatggctgc gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc
     1921 tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt
     1981 tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt
     2041 gaagcgattc acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa
     2101 gcgttaatgt ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg
     2161 tcactgatgc ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac
     2221 gagagaggat gctcacgata cgggttactg atgatgaaca agaggacatc cggtcaaata
     2281 aaacgaaagg ctcagtcgaa agactgggcc tttcgttttg ctgaggagac ttagggactc
     2341 tcacagctaa caccacgtcg tccctatctg ctgccctagg tctatgagtg gttgctggat
     2401 aactttacgg gcatgcataa ggctcgtaat atatattcag ggagaccaca acggtttccc
     2461 tctacaaata attttgttta acttttacta gagtgggacc ctacctgcag gtgcagtact
     2521 acatcatcag gcatgaacac gattaacatc gctaagaacg acttctctga catcgaactg
     2581 gctgctatcc cgttcaacac tctggctgac cattacggtg agcgtttagc tcgcgaacag
     2641 ttggcccttg agcatgagtc ttacgagatg ggtgaagcac gcttccgcaa gatgtttgag
     2701 tgataactta aagctggtga ggttgcggat aacgctgccg ccaagcctct catcactacc
     2761 ctactcccta agatgattgc acgcatcaac gactggtttg aggaagtgaa agctaagcgc
     2821 ggcaagcgcc cgacagcctt ccagttcctg caagaaatca agccggaagc cgtagcgtac
     2881 atcaccatta agaccactct ggcttgccta accagtgctg acaatacaac cgttcaggct
     2941 gtagcaagcg caatcggtcg ggccattgag gacgaggctc gcttcggtcg tatccgtgac
     3001 cttaaagcta agcacttcaa gaaaaacgtt gaggaacaac tcaacaagcg cgtagggcac
     3061 gtctacaaga aagcatttat gcaagttgtc gaggctgaca tgctctctaa gggtctactc
     3121 ggtggcgagg cgtggtcttc gtggcataag gaagactcta ttcatgtagg agtacgctgc
     3181 atcgagatgc tcattgagtc aaccggaatg gttagcttac accgccaaaa tgctggcgta
     3241 gtaggtcaag actctgagac tatcgaactc gcacctgaat acgctgaggc tatcgcaacc
     3301 cgtgcaggtg cgctggctgg catctctccg atgttccaac cttgcgtagt tcctcctaag
     3361 ccgtggactg gcattactgg tggtggctat tgggctaacg gtcgtcgtcc tctggcgctg
     3421 gtgcgtactc acagtaagaa agcactgatg cgctacgaag acgtttacat gcctgaggtg
     3481 tacaaagcga ttaacattgc gcaaaacacc gcatggaaaa tcaacaagaa agtcctagcg
     3541 gtcgccaacg taatcaccaa gtggaagcat tgtccggtcg aggacatccc tgcgattgag
     3601 cgtgaagaac tcccgatgaa accggaagac atcgacatga atcctgaggc tctcaccgcg
     3661 tggaaacgtg ctgccgctgc tgtgtaccgc aaggacaagg ctcgcaagtc tcgccgtatc
     3721 agccttgagt tcatgcttga gcaagccaat aagtttgcta accataaggc catctggttc
     3781 ccttacaaca tggactggcg cggtcgtgtt tacgctgtgt caatgttcaa cccgcaaggt
     3841 aacgatatga ccaaaggact gcttacgctg gcgaaaggta aaccaatcgg taaggaaggt
     3901 tactactggc tgaaaatcca cggtgcaaac tgtgcgggtg tcgataaggt tccgttccct
     3961 gagcgcatca agttcattga ggaaaaccac gagaacatca tggcttgcgc taagtctcca
     4021 ctggagaaca cttggtgggc tgagcaagat tctccgttct gcttccttgc gttctgcttt
     4081 gagtacgctg gggtacagca ccacggcctg agctataact gctcccttcc gctggcgttt
     4141 gacgggtctt gctctggcat ccagcacttc tccgcgatgc tccgagatga ggtaggtggt
     4201 cgcgcggtta acttgcttcc tagtgaaacc gttcaggaca tctacgggat tgttgctaag
     4261 aaagtcaacg agattctaca agcagacgca atcaatggga ccgataacga agtagttacc
     4321 gtgaccgatg agaacactgg tgaaatctct gagaaagtca agctgggcac taaggcactg
     4381 gctggtcaat ggctggctta cggtgttact cgcagtgtga ctaagcgttc agtcatgacg
     4441 ctggcttacg ggtccaaaga gttcggcttc cgtcaacaag tgctggaaga taccattcag
     4501 ccagctattg attccggcaa gggtctgatg ttcactcagc cgaatcaggc tgctggatac
     4561 atggctaagc tgatttggga atctgtgagc gtgacggtgg tagctgcggt tgaagcaatg
     4621 aactggctta agtctgctgc taagctgctg gctgctgagg tcaaagataa gaagactgga
     4681 gagattcttc gcaagcgttg cgctgtgcat tgggtaactc ctgatggttt ccctgtgtgg
     4741 caggaataca agaagcctat tcagacgcgc ttgaacctga tgttcctcgg tcagttccgc
     4801 ttacagccta ccattaacac caacaaagat agcgagattg atgcacacaa acaggagtct
     4861 ggtatcgctc ctaactttgt acacagccaa gacggtagcc accttcgtaa gactgtagtg
     4921 tgggcacacg agaagtacgg tatcgaatct tttgcactga ttcacgactc cttcggtacc
     4981 attccggctg acgctgcgaa cctgttcaaa gcagtgcgcg aaactatggt tgacacatat
     5041 gagtcttgtg atgtactggc tgatttctac gaccagttcg ctgaccagtt gcacgagtct
     5101 caattggaca aaatgccagc acttccggct aaaggtaact tgaacctccg tgacatctta
     5161 gagtccgact tcgcgttcgc ctaatggc
//
