LOCUS       sequence-273047-        5188 bp    DNA     circular SYN 14-APR-2025
DEFINITION  P3e.
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 Apr 14, 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-proD
FEATURES             Location/Qualifiers
     source          1..5188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..56
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     290..307
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    322..462
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(388..407)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     548..570
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(564..755)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3845..4636)
                     /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(4637..4708)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(4736..5188,1..136)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
       61 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      121 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      181 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
      241 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
      301 gagcgaggaa gcggatgagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
      361 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
      421 gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca
      481 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
      541 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg
      601 cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc tgcctgttca
      661 tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct gataaagcgg
      721 gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta agggggattt
      781 ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat acgggttact
      841 gatgatgaac aagaggacat ccggtcaaat aaaacgaaag gctcagtcga aagactgggc
      901 ctttcgtttt gctgaggaga cttagggact ctcacagcta acaccacgtc gtccctatct
      961 gctgccctag gtctatgagt ggttgctgga taactttacg ggcatgcata aggctcgtat
     1021 aatatattca gggagaccac aacggtttcc ctctacaaat aattttgttt aacttttact
     1081 agagtgggac cctacctgca ggtgcagtac tacatcatca ggcatgaaca cgattaacat
     1141 cgctaagaac gacttctctg acatcgaact ggctgctatc ccgttcaaca ctctggctga
     1201 ccattacggt gagcgtttag ctcgcgaaca gttggccctt gagcatgagt cttacgagat
     1261 gggtgaagca cgcttccgca agatgtttga gtgataactt aaagctggtg aggttgcgga
     1321 taacgctgcc gccaagcctc tcatcactac cctactccct aagatgattg cacgcatcaa
     1381 cgactggttt gaggaagtga aagctaagcg cggcaagcgc ccgacagcct tccagttcct
     1441 gcaagaaatc aagccggaag ccgtagcgta catcaccatt aagaccactc tggcttgcct
     1501 aaccagtgct gacaatacaa ccgttcaggc tgtagcaagc gcaatcggtc gggccattga
     1561 ggacgaggct cgcttcggtc gtatccgtga ccttaaagct aagcacttca agaaaaacgt
     1621 tgaggaacaa ctcaacaagc gcgtagggca cgtctacaag aaagcattta tgcaagttgt
     1681 cgaggctgac atgctctcta agggtctact cggtggcgag gcgtggtctt cgtggcataa
     1741 ggaagactct attcatgtag gagtacgctg catcgagatg ctcattgagt caaccggaat
     1801 ggttagctta caccgccaaa atgctggcgt agtaggtcaa gactctgaga ctatcgaact
     1861 cgcacctgaa tacgctgagg ctatcgcaac ccgtgcaggt gcgctggctg gcatctctcc
     1921 gatgttccaa ccttgcgtag ttcctcctaa gccgtggact ggcattactg gtggtggcta
     1981 ttgggctaac ggtcgtcgtc ctctggcgct ggtgcgtact cacagtaaga aagcactgat
     2041 gcgctacgaa gacgtttaca tgcctgaggt gtacaaagcg attaacattg cgcaaaacac
     2101 cgcatggaaa atcaacaaga aagtcctagc ggtcgccaac gtaatcacca agtggaagca
     2161 ttgtccggtc gaggacatcc ctgcgattga gcgtgaagaa ctcccgatga aaccggaaga
     2221 catcgacatg aatcctgagg ctctcaccgc gtggaaacgt gctgccgctg ctgtgtaccg
     2281 caaggacaag gctcgcaagt ctcgccgtat cagccttgag ttcatgcttg agcaagccaa
     2341 taagtttgct aaccataagg ccatctggtt cccttacaac atggactggc gcggtcgtgt
     2401 ttacgctgtg tcaatgttca acccgcaagg taacgatatg accaaaggac tgcttacgct
     2461 ggcgaaaggt aaaccaatcg gtaaggaagg ttactactgg ctgaaaatcc acggtgcaaa
     2521 ctgtgcgggt gtcgataagg ttccgttccc tgagcgcatc aagttcattg aggaaaacca
     2581 cgagaacatc atggcttgcg ctaagtctcc actggagaac acttggtggg ctgagcaaga
     2641 ttctccgttc tgcttccttg cgttctgctt tgagtacgct ggggtacagc accacggcct
     2701 gagctataac tgctcccttc cgctggcgtt tgacgggtct tgctctggca tccagcactt
     2761 ctccgcgatg ctccgagatg aggtaggtgg tcgcgcggtt aacttgcttc ctagtgaaac
     2821 cgttcaggac atctacggga ttgttgctaa gaaagtcaac gagattctac aagcagacgc
     2881 aatcaatggg accgataacg aagtagttac cgtgaccgat gagaacactg gtgaaatctc
     2941 tgagaaagtc aagctgggca ctaaggcact ggctggtcaa tggctggctt acggtgttac
     3001 tcgcagtgtg actaagcgtt cagtcatgac gctggcttac gggtccaaag agttcggctt
     3061 ccgtcaacaa gtgctggaag ataccattca gccagctatt gattccggca agggtctgat
     3121 gttcactcag ccgaatcagg ctgctggata catggctaag ctgatttggg aatctgtgag
     3181 cgtgacggtg gtagctgcgg ttgaagcaat gaactggctt aagtctgctg ctaagctgct
     3241 ggctgctgag gtcaaagata agaagactgg agagattctt cgcaagcgtt gcgctgtgca
     3301 ttgggtaact cctgatggtt tccctgtgtg gcaggaatac aagaagccta ttcagacgcg
     3361 cttgaacctg atgttcctcg gtcagttccg cttacagcct accattaaca ccaacaaaga
     3421 tagcgagatt gatgcacaca aacaggagtc tggtatcgct cctaactttg tacacagcca
     3481 agacggtagc caccttcgta agactgtagt gtgggcacac gagaagtacg gtatcgaatc
     3541 ttttgcactg attcacgact ccttcggtac cattccggct gacgctgcga acctgttcaa
     3601 agcagtgcgc gaaactatgg ttgacacata tgagtcttgt gatgtactgg ctgatttcta
     3661 cgaccagttc gctgaccagt tgcacgagtc tcaattggac aaaatgccag cacttccggc
     3721 taaaggtaac ttgaacctcc gtgacatctt agagtccgac ttcgcgttcg cctaatggca
     3781 ctcctcagca aatataatga ccctcttgat aacccaagag ggcatttttt aatgcccatg
     3841 gcgtttattt gccgactacc ttggtgatct cgcctttcac gtagtggaca aattcttcca
     3901 actgatctgc gcgcgaggcc aagcgatctt cttcttgtcc aagataagcc tgtctagctt
     3961 caagtatgac gggctgatac tgggccggca ggcgctccat tgcccagtcg gcagcgacat
     4021 ccttcggcgc gattttgccg gttactgcgc tgtaccaaat gcgggacaac gtaagcacta
     4081 catttcgctc atcgccagcc cagtcgggcg gcgagttcca tagcgttaag gtttcattta
     4141 gcgcctcaaa tagatcctgt tcaggaaccg gatcaaagag ttcctccgcc gctggaccta
     4201 ccaaggcaac gctatgttct cttgcttttg tcagcaagat agccagatca atgtcgatcg
     4261 tggctggctc gaagatacct gcaagaatgt cattgcgctg ccattctcca aattgcagtt
     4321 cgcgcttagc tggataacgc cacggaatga tgtcgtcgtg cacaacaatg gtgacttcta
     4381 cagcgcggag aatctcgctc tctccagggg aagccgaagt ttccaaaagg tcgttgatca
     4441 aagctcgccg cgttgtttca tcaagcctta cggtcaccgt aaccagcaaa tcaatatcac
     4501 tgtgtggctt caggccgcca tccactgcgg agccgtacaa atgtacggcc agcaacgtcg
     4561 gttcgagatg gcgctcgatg acgccaacta cctctgatag ttgagtcgat acttcggcga
     4621 tcaccgcttc cctcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4681 gtctcatgag cgtatacata tttgaatgta tttagaaaaa taggccaaat agggtttgag
     4741 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     4801 tggtttgttt gccggatcaa gagctaccac ctctttttcc gaaggtaact ggcttcagca
     4861 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     4921 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4981 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5041 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5101 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5161 aggcggacag gtatccggta agcggcag
//
