LOCUS       sequence-352078-        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: pBM13g-sd4U
FEATURES             Location/Qualifiers
     source          1..6093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      43..88
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(103..981)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1008..1292
                     /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     1215..1234
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             complement(3860..4651)
                     /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(4652..4723)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4751..5339
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5240..5259
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5493..5510
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5525..5665
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5591..5610)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5751..5773
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5767..5958)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aagactgggc ctttcgtttt gctgaggaga cttagggact ctcaaataaa acgaaaggct
       61 cagtcgaaag actgggcctt tcgttttaga cttagggacc ctttatgaca acttgacggc
      121 tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg ccccggtgca
      181 ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc gaccgacggt
      241 ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac gttggtcctc
      301 gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca gacgcgacgg
      361 cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg ccaggtgatc
      421 gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga gcgactcgtt
      481 aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca gcagctccga
      541 atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc tgaaatgcgg
      601 ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg gccagttaag
      661 ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc attcgcgagc
      721 ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa tatcacccgg
      781 tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa tggtgagatt
      841 gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat aaccgttggc
      901 ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg gcagcagggg
      961 atcattttgc gcttcagcca tacttttcat actcccacca ttcagagaag aaaccaattg
     1021 tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct cgctaaccca
     1081 accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag ccatgacaaa
     1141 aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt atttgcacgg
     1201 cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc ctacctgacg
     1261 ctttttatcg caactctcta ctgtttctcc atacccgttt ttttggacgc gtacaactca
     1321 agtctgacaa aaaaaaggaa aaattgacca acctgtctga catcatcgag aaggagacag
     1381 gcaagcagct ggtcatccag gagagcatcc tgatgctgcc cgaagaagtc gaagaagtga
     1441 tcggaaacaa gcctgagagc gatatcctgg tccataccgc ctacaacgag agtaccgatg
     1501 aaaatgtgat gctgctgaca tccaacgccc cagagtataa gccctgggct ctggtcatcc
     1561 aggattccaa cggagagaac aaaatcaaaa tgctgtctgg tggttctgac atcgctgatc
     1621 tgagaaccct gggctactcc cagcagcaac aggagaagat caagcctaag gtgcggagca
     1681 cagtcgccca gcaccacgag gccctggtgg gccacggctt cacccacgcc cacatcgtgg
     1741 ccctcagcca gcatcctgcc gccctgggga ccgtggccgt gaagtaccag gacatgatcg
     1801 cggctctgcc agaagccacc cacgaagcca tcgtgggcgt gggcaagcaa tggtccggcg
     1861 ccagagccct ggaggccctg ctgaccgtgg ccggtgagct gagaggccct cctctgcagc
     1921 tggacaccgg ccagctgctc aagatcgcta agcggggcgg agtaaccgct gtcgaggccg
     1981 tgcacgcctg gcggaacgcc ctaaccggag cccctctaaa cctgacccca gagcaggtcg
     2041 tggcaatcgc aagccacgat ggcggcaagc aggccctgga aacagtgcag gccctgctac
     2101 ctgtcctgtg ccaggcccac gaactgaccc ccaagcaggt ggtagccatc gccagccacg
     2161 atggcggaaa acaggccctg aaaacagtgc agcggctgct gcccgtgctg tgccaggccc
     2221 acggcctgac ccctcaacag gtcgtggcca tcgctagcaa caacggcggc aagcaggccc
     2281 tggagactgt gcagagattg ctgcctgtgc tgtgccaggc ccacggcctg acccctgagc
     2341 aggtggtggc tatcgcctct aacatcggcg gcaagcaggc cctggagacc gttcaggcac
     2401 tgctacctgt tctgtgtcag gctcacggac tgacacctga gcaggtggtg gcaatcgcca
     2461 gccacgacgg cggcaagcag gccttggaaa cagtgcaggc cctgctgcca gtgctgtgcc
     2521 aggcccacgg cctgacccct gagcaggtgg tagccattgc cagcaacggc ggaggcaaac
     2581 aagccctgga aaccgtgcag gctctgctgc ctgtgctgtg tcaagcccat ggcctcacac
     2641 ccgagcaggt ggtggctatc gcctccaatg gaggcggcaa acaggccctc gagaccgtgc
     2701 agagactgct gcctgtgctg tgccaggccc acggcctgac acccgaacaa gttgtggcaa
     2761 tcgcctctca cgacggcgga aaacaggccc tggaaaccgt gcaggctctt ctgcccgtgc
     2821 tgtgtcaggc ccacgaactg acccccgagc aggtggtggc tatcgcaagc aacaacggcg
     2881 gcaagcaggc cctggaaaca gtccaaaggc tgctgcccgt gctgtgccag gcccacggcc
     2941 tgacacctga gcaggtggtg gccatcgcca gccatgacgg cggcaagcag gcacttgaaa
     3001 ccgttcaggc tctgctgccc gtcctgtgcc aggcccacgg cctaacccct gaacaagtgg
     3061 tggccatcgc ctcaaacaat ggcggcaagc aagccctgaa aaccgtgcag gccctgctgc
     3121 ctgtcctgtg ccaggcccac ggactgaccc ccgagcaggt ggtagccatc gcatctaatg
     3181 gcggagggaa gcaggctctg gagacggtgc aagcactcct gcccgtgctg tgccaggccc
     3241 acggcctgac cccccagcaa gtggttgcta tcgccagcaa cggcggcggt agaccagccc
     3301 tggagagcat tgtggcccag ctgagcagac ctgatcctgc cctggccgcc ctgaccaacg
     3361 accacctggt ggccctggcc tgcctgggcg gaagacccgc tttgaacgcc gtgaaaaagg
     3421 gcctgggtgg atccggcagc tacgccctgg gtccgtatca gattagcgcc ccgcagctgc
     3481 cagcttacaa tggtcagacc gtgggtacct tctactatgt gaacgacgcg ggcggtctgg
     3541 agagcaaggt gtttatcagc ggcggtccaa ccccgtaccc aaactatgtc agtgccggtc
     3601 atgtggaggg tcagagcgcc ctgttcatgc gtgataacgg catcagcgag ggtctggtgt
     3661 tccacaacaa cccgaaaggc acctgcggtt tttgcgtgaa catgatcgag accctgctgc
     3721 cggaaaacgc gaaaatgacc gtggtgccgc cggaaggtta ataaacttaa ttaacggcac
     3781 tcctcagcaa aggcactcct cagcaaatat aatgaccctc ttgataaccc aagagggcat
     3841 tttttaatgc ccatggcgtt tatttgccga ctaccttggt gatctcgcct ttcacgtagt
     3901 ggacaaattc ttccaactga tctgcgcgcg aggccaagcg atcttcttct tgtccaagat
     3961 aagcctgtct agcttcaagt atgacgggct gatactgggc cggcaggcgc tccattgccc
     4021 agtcggcagc gacatccttc ggcgcgattt tgccggttac tgcgctgtac caaatgcggg
     4081 acaacgtaag cactacattt cgctcatcgc cagcccagtc gggcggcgag ttccatagcg
     4141 ttaaggtttc atttagcgcc tcaaatagat cctgttcagg aaccggatca aagagttcct
     4201 ccgccgctgg acctaccaag gcaacgctat gttctcttgc ttttgtcagc aagatagcca
     4261 gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag aatgtcattg cgctgccatt
     4321 ctccaaattg cagttcgcgc ttagctggat aacgccacgg aatgatgtcg tcgtgcacaa
     4381 caatggtgac ttctacagcg cggagaatct cgctctctcc aggggaagcc gaagtttcca
     4441 aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag ccttacggtc accgtaacca
     4501 gcaaatcaat atcactgtgt ggcttcaggc cgccatccac tgcggagccg tacaaatgta
     4561 cggccagcaa cgtcggttcg agatggcgct cgatgacgcc aactacctct gatagttgag
     4621 tcgatacttc ggcgatcacc gcttccctca tactcttcct ttttcaatat tattgaagca
     4681 tttatcaggg ttattgtctc atgagcgtat acatatttga atgtatttag aaaaataggc
     4741 caaatagggt ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4801 caccgctacc agcggtggtt tgtttgccgg atcaagagct accacctctt tttccgaagg
     4861 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     4921 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4981 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5041 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5101 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5161 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagagtcgga acaggagagc
     5221 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5281 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5341 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     5401 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     5461 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggatg
     5521 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt
     5581 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc
     5641 gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg
     5701 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     5761 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc
     5821 atcagcgtgg tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt
     5881 gagtttctcc agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt
     5941 tttttcctgt ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat
     6001 gataccgatg aaacgagaga ggatgctcac gatacgggtt actgatgatg aacaagagga
     6061 catccggtca aataaaacga aaggctcagt cga
//
