LOCUS       sequence-355096-        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: pBM13-sd4U
FEATURES             Location/Qualifiers
     source          1..6093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      134..179
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(194..1072)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1099..1383
                     /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     1306..1325
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             complement(3951..4742)
                     /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(4743..4814)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4842..5430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5331..5350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5584..5601
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5616..5756
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5682..5701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5842..5864
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5858..6049)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa caagaggaca
       61 tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tgctgaggag
      121 acttagggac tctcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttag
      181 acttagggac cctttatgac aacttgacgg ctacatcatt cactttttct tcacaaccgg
      241 cacggaactc gctcgggctg gccccggtgc attttttaaa tacccgcgag aaatagagtt
      301 gatcgtcaaa accaacattg cgaccgacgg tggcgatagg catccgggtg gtgctcaaaa
      361 gcagcttcgc ctggctgata cgttggtcct cgcgccagct taagacgcta atccctaact
      421 gctggcggaa aagatgtgac agacgcgacg gcgacaagca aacatgctgt gcgacgctgg
      481 cgatatcaaa attgctgtct gccaggtgat cgctgatgta ctgacaagcc tcgcgtaccc
      541 gattatccat cggtggatgg agcgactcgt taatcgcttc catgcgccgc agtaacaatt
      601 gctcaagcag atttatcgcc agcagctccg aatagcgccc ttccccttgc ccggcgttaa
      661 tgatttgccc aaacaggtcg ctgaaatgcg gctggtgcgc ttcatccggg cgaaagaacc
      721 ccgtattggc aaatattgac ggccagttaa gccattcatg ccagtaggcg cgcggacgaa
      781 agtaaaccca ctggtgatac cattcgcgag cctccggatg acgaccgtag tgatgaatct
      841 ctcctggcgg gaacagcaaa atatcacccg gtcggcaaac aaattctcgt ccctgatttt
      901 tcaccacccc ctgaccgcga atggtgagat tgagaatata acctttcatt cccagcggtc
      961 ggtcgataaa aaaatcgaga taaccgttgg cctcaatcgg cgttaaaccc gccaccagat
     1021 gggcattaaa cgagtatccc ggcagcaggg gatcattttg cgcttcagcc atacttttca
     1081 tactcccacc attcagagaa gaaaccaatt gtccatattg catcagacat tgccgtcact
     1141 gcgtctttta ctggctcttc tcgctaaccc aaccggtaac cccgcttatt aaaagcattc
     1201 tgtaacaaag cgggaccaaa gccatgacaa aaacgcgtaa caaaagtgtc tataatcacg
     1261 gcagaaaagt ccacattgat tatttgcacg gcgtcacact ttgctatgcc atagcatttt
     1321 tatccataag attagcggat cctacctgac gctttttatc gcaactctct actgtttctc
     1381 catacccgtt tttttggacg cgtacaactc aagtctgaca aaaaaaagga aaaattgacc
     1441 aacctgtctg acatcatcga gaaggagaca ggcaagcagc tggtcatcca ggagagcatc
     1501 ctgatgctgc ccgaagaagt cgaagaagtg atcggaaaca agcctgagag cgatatcctg
     1561 gtccataccg cctacaacga gagtaccgat gaaaatgtga tgctgctgac atccaacgcc
     1621 ccagagtata agccctgggc tctggtcatc caggattcca acggagagaa caaaatcaaa
     1681 atgctgtctg gtggttctga catcgctgat ctgagaaccc tgggctactc ccagcagcaa
     1741 caggagaaga tcaagcctaa ggtgcggagc acagtcgccc agcaccacga ggccctggtg
     1801 ggccacggct tcacccacgc ccacatcgtg gccctcagcc agcatcctgc cgccctgggg
     1861 accgtggccg tgaagtacca ggacatgatc gcggctctgc cagaagccac ccacgaagcc
     1921 atcgtgggcg tgggcaagca atggtccggc gccagagccc tggaggccct gctgaccgtg
     1981 gccggtgagc tgagaggccc tcctctgcag ctggacaccg gccagctgct caagatcgct
     2041 aagcggggcg gagtaaccgc tgtcgaggcc gtgcacgcct ggcggaacgc cctaaccgga
     2101 gcccctctaa acctgacccc agagcaggtc gtggcaatcg caagccacga tggcggcaag
     2161 caggccctgg aaacagtgca ggccctgcta cctgtcctgt gccaggccca cgaactgacc
     2221 cccaagcagg tggtagccat cgccagccac gatggcggaa aacaggccct gaaaacagtg
     2281 cagcggctgc tgcccgtgct gtgccaggcc cacggcctga cccctcaaca ggtcgtggcc
     2341 atcgctagca acaacggcgg caagcaggcc ctggagactg tgcagagatt gctgcctgtg
     2401 ctgtgccagg cccacggcct gacccctgag caggtggtgg ctatcgcctc taacatcggc
     2461 ggcaagcagg ccctggagac cgttcaggca ctgctacctg ttctgtgtca ggctcacgga
     2521 ctgacacctg agcaggtggt ggcaatcgcc agccacgacg gcggcaagca ggccttggaa
     2581 acagtgcagg ccctgctgcc agtgctgtgc caggcccacg gcctgacccc tgagcaggtg
     2641 gtagccattg ccagcaacgg cggaggcaaa caagccctgg aaaccgtgca ggctctgctg
     2701 cctgtgctgt gtcaagccca tggcctcaca cccgagcagg tggtggctat cgcctccaat
     2761 ggaggcggca aacaggccct cgagaccgtg cagagactgc tgcctgtgct gtgccaggcc
     2821 cacggcctga cacccgaaca agttgtggca atcgcctctc acgacggcgg aaaacaggcc
     2881 ctggaaaccg tgcaggctct tctgcccgtg ctgtgtcagg cccacgaact gacccccgag
     2941 caggtggtgg ctatcgcaag caacaacggc ggcaagcagg ccctggaaac agtccaaagg
     3001 ctgctgcccg tgctgtgcca ggcccacggc ctgacacctg agcaggtggt ggccatcgcc
     3061 agccatgacg gcggcaagca ggcacttgaa accgttcagg ctctgctgcc cgtcctgtgc
     3121 caggcccacg gcctaacccc tgaacaagtg gtggccatcg cctcaaacaa tggcggcaag
     3181 caagccctga aaaccgtgca ggccctgctg cctgtcctgt gccaggccca cggactgacc
     3241 cccgagcagg tggtagccat cgcatctaat ggcggaggga agcaggctct ggagacggtg
     3301 caagcactcc tgcccgtgct gtgccaggcc cacggcctga ccccccagca agtggttgct
     3361 atcgccagca acggcggcgg tagaccagcc ctggagagca ttgtggccca gctgagcaga
     3421 cctgatcctg ccctggccgc cctgaccaac gaccacctgg tggccctggc ctgcctgggc
     3481 ggaagacccg ctttgaacgc cgtgaaaaag ggcctgggtg gatccggcag ctacgccctg
     3541 ggtccgtatc agattagcgc cccgcagctg ccagcttaca atggtcggac cgtgggtacc
     3601 ttctactatg tgaacgacgc gggcggtctg gagagcaagg tgtttatcag cggcggtcca
     3661 accccgtacc caaactatgc caatgccggt catgtggagg gtcagagcgc cctgttcatg
     3721 cgtgataacg gcatcagcga gggtctggtg ttccacaaca acccggaagg cacctgcggt
     3781 ttttgcgtga acatgatcga gaccctgctg ccggaaaacg cgaaaatgac cgtggtgccg
     3841 ccggaaggtt aataaactta attaacggca ctcctcagca aaggcactcc tcagcaaata
     3901 taatgaccct cttgataacc caagagggca ttttttaatg cccatggcgt ttatttgccg
     3961 actaccttgg tgatctcgcc tttcacgtag tggacaaatt cttccaactg atctgcgcgc
     4021 gaggccaagc gatcttcttc ttgtccaaga taagcctgtc tagcttcaag tatgacgggc
     4081 tgatactggg ccggcaggcg ctccattgcc cagtcggcag cgacatcctt cggcgcgatt
     4141 ttgccggtta ctgcgctgta ccaaatgcgg gacaacgtaa gcactacatt tcgctcatcg
     4201 ccagcccagt cgggcggcga gttccatagc gttaaggttt catttagcgc ctcaaataga
     4261 tcctgttcag gaaccggatc aaagagttcc tccgccgctg gacctaccaa ggcaacgcta
     4321 tgttctcttg cttttgtcag caagatagcc agatcaatgt cgatcgtggc tggctcgaag
     4381 atacctgcaa gaatgtcatt gcgctgccat tctccaaatt gcagttcgcg cttagctgga
     4441 taacgccacg gaatgatgtc gtcgtgcaca acaatggtga cttctacagc gcggagaatc
     4501 tcgctctctc caggggaagc cgaagtttcc aaaaggtcgt tgatcaaagc tcgccgcgtt
     4561 gtttcatcaa gccttacggt caccgtaacc agcaaatcaa tatcactgtg tggcttcagg
     4621 ccgccatcca ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc
     4681 tcgatgacgc caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc
     4741 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgta
     4801 tacatatttg aatgtattta gaaaaatagg ccaaataggg tttgagatcc tttttttctg
     4861 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4921 gatcaagagc taccacctct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4981 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5041 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5101 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5161 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5221 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5281 ccggtaagcg gcagagtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5341 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5401 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5461 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5521 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5581 cgcagcgagt cagtgagcga ggaagcggat gagcgcctga tgcggtattt tctccttacg
     5641 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     5701 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     5761 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
     5821 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     5881 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
     5941 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
     6001 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
     6061 tgtaaggggg atttctgttc atgggggtaa tga
//
