LOCUS       sequence-355095-        6093 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of base editor .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6093)
  AUTHORS   Mok BY, Kotrys AV, Raguram A, Huang TP, Mootha VK, Liu DR
  TITLE     CRISPR-free base editors with enhanced activity and expanded 
            targeting scope in mitochondrial and nuclear DNA.
  JOURNAL   Nat Biotechnol. 2022 Sep;40(9):1378-1387. doi: 
            10.1038/s41587-022-01256-8. Epub 2022 Apr 4.
  PUBMED    35379961
REFERENCE   2  (bases 1 to 6093)
  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/s41587-022-01256-8"; journalName: "Nat Biotechnol"; date: 
            "2022-09"; volume: "40"; issue: "9"; pages: "1378-1387"
COMMENT     Sequence Label: pBM13h-sd4U
FEATURES             Location/Qualifiers
     source          1..6093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      155..200
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(215..1093)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1120..1404
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1327..1346
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             complement(3972..4763)
                     /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(4764..4835)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4863..5451
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5352..5371
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5605..5622
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5637..5777
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5703..5722)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5863..5885
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5879..6070)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
       61 ctgatgatga acaagaggac atccggtcaa ataaaacgaa aggctcagtc gaaagactgg
      121 gcctttcgtt ttgctgagga gacttaggga ctctcaaata aaacgaaagg ctcagtcgaa
      181 agactgggcc tttcgtttta gacttaggga ccctttatga caacttgacg gctacatcat
      241 tcactttttc ttcacaaccg gcacggaact cgctcgggct ggccccggtg cattttttaa
      301 atacccgcga gaaatagagt tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag
      361 gcatccgggt ggtgctcaaa agcagcttcg cctggctgat acgttggtcc tcgcgccagc
      421 ttaagacgct aatccctaac tgctggcgga aaagatgtga cagacgcgac ggcgacaagc
      481 aaacatgctg tgcgacgctg gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt
      541 actgacaagc ctcgcgtacc cgattatcca tcggtggatg gagcgactcg ttaatcgctt
      601 ccatgcgccg cagtaacaat tgctcaagca gatttatcgc cagcagctcc gaatagcgcc
      661 cttccccttg cccggcgtta atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg
      721 cttcatccgg gcgaaagaac cccgtattgg caaatattga cggccagtta agccattcat
      781 gccagtaggc gcgcggacga aagtaaaccc actggtgata ccattcgcga gcctccggat
      841 gacgaccgta gtgatgaatc tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa
      901 caaattctcg tccctgattt ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat
      961 aacctttcat tcccagcggt cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg
     1021 gcgttaaacc cgccaccaga tgggcattaa acgagtatcc cggcagcagg ggatcatttt
     1081 gcgcttcagc catacttttc atactcccac cattcagaga agaaaccaat tgtccatatt
     1141 gcatcagaca ttgccgtcac tgcgtctttt actggctctt ctcgctaacc caaccggtaa
     1201 ccccgcttat taaaagcatt ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta
     1261 acaaaagtgt ctataatcac ggcagaaaag tccacattga ttatttgcac ggcgtcacac
     1321 tttgctatgc catagcattt ttatccataa gattagcgga tcctacctga cgctttttat
     1381 cgcaactctc tactgtttct ccatacccgt ttttttggac gcgtacaact caagtctgac
     1441 aaaaaaaagg aaaaattgac caacctgtct gacatcatcg agaaggagac aggcaagcag
     1501 ctggtcatcc aggagagcat cctgatgctg cccgaagaag tcgaagaagt gatcggaaac
     1561 aagcctgaga gcgatatcct ggtccatacc gcctacaacg agagtaccga tgaaaatgtg
     1621 atgctgctga catccaacgc cccagagtat aagccctggg ctctggtcat ccaggattcc
     1681 aacggagaga acaaaatcaa aatgctgtct ggtggttctg acatcgctga tctgagaacc
     1741 ctgggctact cccagcagca acaggagaag atcaagccta aggtgcggag cacagtcgcc
     1801 cagcaccacg aggccctggt gggccacggc ttcacccacg cccacatcgt ggccctcagc
     1861 cagcatcctg ccgccctggg gaccgtggcc gtgaagtacc aggacatgat cgcggctctg
     1921 ccagaagcca cccacgaagc catcgtgggc gtgggcaagc aatggtccgg cgccagagcc
     1981 ctggaggccc tgctgaccgt ggccggtgag ctgagaggcc ctcctctgca gctggacacc
     2041 ggccagctgc tcaagatcgc taagcggggc ggagtaaccg ctgtcgaggc cgtgcacgcc
     2101 tggcggaacg ccctaaccgg agcccctcta aacctgaccc cagagcaggt cgtggcaatc
     2161 gcaagccacg atggcggcaa gcaggccctg gaaacagtgc aggccctgct acctgtcctg
     2221 tgccaggccc acgaactgac ccccaagcag gtggtagcca tcgccagcca cgatggcgga
     2281 aaacaggccc tgaaaacagt gcagcggctg ctgcccgtgc tgtgccaggc ccacggcctg
     2341 acccctcaac aggtcgtggc catcgctagc aacaacggcg gcaagcaggc cctggagact
     2401 gtgcagagat tgctgcctgt gctgtgccag gcccacggcc tgacccctga gcaggtggtg
     2461 gctatcgcct ctaacatcgg cggcaagcag gccctggaga ccgttcaggc actgctacct
     2521 gttctgtgtc aggctcacgg actgacacct gagcaggtgg tggcaatcgc cagccacgac
     2581 ggcggcaagc aggccttgga aacagtgcag gccctgctgc cagtgctgtg ccaggcccac
     2641 ggcctgaccc ctgagcaggt ggtagccatt gccagcaacg gcggaggcaa acaagccctg
     2701 gaaaccgtgc aggctctgct gcctgtgctg tgtcaagccc atggcctcac acccgagcag
     2761 gtggtggcta tcgcctccaa tggaggcggc aaacaggccc tcgagaccgt gcagagactg
     2821 ctgcctgtgc tgtgccaggc ccacggcctg acacccgaac aagttgtggc aatcgcctct
     2881 cacgacggcg gaaaacaggc cctggaaacc gtgcaggctc ttctgcccgt gctgtgtcag
     2941 gcccacgaac tgacccccga gcaggtggtg gctatcgcaa gcaacaacgg cggcaagcag
     3001 gccctggaaa cagtccaaag gctgctgccc gtgctgtgcc aggcccacgg cctgacacct
     3061 gagcaggtgg tggccatcgc cagccatgac ggcggcaagc aggcacttga aaccgttcag
     3121 gctctgctgc ccgtcctgtg ccaggcccac ggcctaaccc ctgaacaagt ggtggccatc
     3181 gcctcaaaca atggcggcaa gcaagccctg aaaaccgtgc aggccctgct gcctgtcctg
     3241 tgccaggccc acggactgac ccccgagcag gtggtagcca tcgcatctaa tggcggaggg
     3301 aagcaggctc tggagacggt gcaagcactc ctgcccgtgc tgtgccaggc ccacggcctg
     3361 accccccagc aagtggttgc tatcgccagc aacggcggcg gtagaccagc cctggagagc
     3421 attgtggccc agctgagcag acctgatcct gccctggccg ccctgaccaa cgaccacctg
     3481 gtggccctgg cctgcctggg cggaagaccc gctttgaacg ccgtgaaaaa gggcctgggt
     3541 ggatccggca gctacgccct gggtccgtat cagattagcg ccccgcagct gccagcttac
     3601 aatggtcaga ccgtgggtac cttctactat gtgaacgacg cgggcggtct ggagagcaag
     3661 gtgtttagca gcggcggtcc aaccccgtac ccaaactatg ccaatgccgg tcatgtggag
     3721 ggtcagagcg ccctgttcat gcgtgataac ggcatcagcg agggtctggt gttccacaac
     3781 aacccggaag gcacctgcgg tttttgcgtg aacatgaccg agaccctgct gccggaaaac
     3841 gcgaaaatga ccgtggtgcc gccggaaggt taataaactt aattaacggc actcctcagc
     3901 aaaggcactc ctcagcaaat ataatgaccc tcttgataac ccaagagggc attttttaat
     3961 gcccatggcg tttatttgcc gactaccttg gtgatctcgc ctttcacgta gtggacaaat
     4021 tcttccaact gatctgcgcg cgaggccaag cgatcttctt cttgtccaag ataagcctgt
     4081 ctagcttcaa gtatgacggg ctgatactgg gccggcaggc gctccattgc ccagtcggca
     4141 gcgacatcct tcggcgcgat tttgccggtt actgcgctgt accaaatgcg ggacaacgta
     4201 agcactacat ttcgctcatc gccagcccag tcgggcggcg agttccatag cgttaaggtt
     4261 tcatttagcg cctcaaatag atcctgttca ggaaccggat caaagagttc ctccgccgct
     4321 ggacctacca aggcaacgct atgttctctt gcttttgtca gcaagatagc cagatcaatg
     4381 tcgatcgtgg ctggctcgaa gatacctgca agaatgtcat tgcgctgcca ttctccaaat
     4441 tgcagttcgc gcttagctgg ataacgccac ggaatgatgt cgtcgtgcac aacaatggtg
     4501 acttctacag cgcggagaat ctcgctctct ccaggggaag ccgaagtttc caaaaggtcg
     4561 ttgatcaaag ctcgccgcgt tgtttcatca agccttacgg tcaccgtaac cagcaaatca
     4621 atatcactgt gtggcttcag gccgccatcc actgcggagc cgtacaaatg tacggccagc
     4681 aacgtcggtt cgagatggcg ctcgatgacg ccaactacct ctgatagttg agtcgatact
     4741 tcggcgatca ccgcttccct catactcttc ctttttcaat attattgaag catttatcag
     4801 ggttattgtc tcatgagcgt atacatattt gaatgtattt agaaaaatag gccaaatagg
     4861 gtttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4921 ccagcggtgg tttgtttgcc ggatcaagag ctaccacctc tttttccgaa ggtaactggc
     4981 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     5041 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5101 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5161 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5221 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5281 gggagaaagg cggacaggta tccggtaagc ggcagagtcg gaacaggaga gcgcacgagg
     5341 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5401 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5461 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5521 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     5581 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga tgagcgcctg
     5641 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     5701 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     5761 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5821 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5881 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     5941 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     6001 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     6061 gtttggtcac tgatgcctcc gtgtaagggg gat
//
