LOCUS       sequence-273887-        7869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A-to-G base editor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7869)
  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 7869)
  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: SaKKH-ABE8e-dimer
FEATURES             Location/Qualifiers
     source          1..7869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        132..335
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     286..306
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     377..396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        377..395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             445..465
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4855..4875
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4884..4901
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4924..4941)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4930..5154
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5225..5241)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5225..5241)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5238..5260)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5249..5265
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5273..5279,5280..5297,5298..5303))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5318..5339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5456..5473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5627..6215)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5707..5726)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6386..7177,7178..7246))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7009..7028
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7247..7351)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7430..7449)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        join(7621..7869,1..131)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
       61 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      121 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      181 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      241 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      301 ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct
      361 agagatccgc ggccgctaat acgactcact atagggagag ccgccaccat gaaacggaca
      421 gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtctctga agtcgagttt
      481 agccacgagt attggatgag gcacgcactg accctggcaa agcgagcatg ggatgaaaga
      541 gaagtccccg tgggcgccgt gctggtgcac aacaatagag tgatcggaga gggatggaac
      601 aggccaatcg gccgccacga ccctaccgca cacgcagaga tcatggcact gaggcaggga
      661 ggcctggtca tgcagaatta ccgcctgatc gatgccaccc tgtatgtgac actggagcca
      721 tgcgtgatgt gcgcaggagc aatgatccac agcaggatcg gaagagtggt gttcggagca
      781 cgggacgcca agaccggcgc agcaggctcc ctgatggatg tgctgcacca ccccggcatg
      841 aaccaccggg tggagatcac agagggaatc ctggcagacg agtgcgccgc cctgctgagc
      901 gatttcttta gaatgcggag acaggagatc aaggcccaga agaaggcaca gagctccacc
      961 gactctggag gatctagcgg aggatcctct ggaagcgaga caccaggcac aagcgagtcc
     1021 gccacaccag agagctccgg cggctcctcc ggaggatcct ctgaggtgga gttttcccac
     1081 gagtactgga tgagacatgc cctgaccctg gccaagaggg cacgggatga gagggaggtg
     1141 cctgtgggag ccgtgctggt gctgaacaat agagtgatcg gcgagggctg gaacagagcc
     1201 atcggcctgc acgacccaac agcccatgcc gaaattatgg ccctgagaca gggcggcctg
     1261 gtcatgcaga actacagact gattgacgcc accctgtacg tgacattcga gccttgcgtg
     1321 atgtgcgccg gcgccatgat ccactctagg atcggccgcg tggtgtttgg cgtgaggaac
     1381 tcaaaaagag gcgccgcagg ctccctgatg aacgtgctga actaccccgg catgaatcac
     1441 cgcgtcgaaa ttaccgaggg aatcctggca gatgaatgtg ccgccctgct gtgcgatttc
     1501 tatcggatgc ctagacaggt gttcaatgct cagaagaagg cccagagctc catcaactcc
     1561 ggaggatcta gcggaggctc ctctggctct gagacacctg gcacaagcga gagcgcaaca
     1621 cctgaaagca gcgggggcag cagcgggggg tcagggaagc gaaattacat tctggggctg
     1681 gccattggca ttacatcagt gggctatggc atcattgact acgagacaag ggacgtgatc
     1741 gacgccggcg tgagactgtt caaggaggcc aacgtggaga acaatgaggg ccggagatcc
     1801 aagaggggag caaggcgcct gaagcggaga aggcgccaca gaatccagag agtgaagaag
     1861 ctgctgttcg attacaacct gctgaccgac cactccgagc tgtctggcat caatccttat
     1921 gaggccagag tgaagggcct gtcccagaag ctgtctgagg aggagtttag cgccgccctg
     1981 ctgcacctgg caaagaggag aggcgtgcac aacgtgaatg aggtggagga ggacaccggc
     2041 aacgagctgt ccacaaagga gcagatcagc cgcaattcca aggccctgga ggagaagtat
     2101 gtggccgagc tgcagctgga gcggctgaag aaggatggcg aggtgagggg ctccatcaat
     2161 cgcttcaaga cctctgacta cgtgaaggag gccaagcagc tgctgaaggt gcagaaggcc
     2221 taccaccagc tggatcagtc ctttatcgat acatatatcg acctgctgga gacaaggcgc
     2281 acatactatg agggaccagg agagggctct cccttcggct ggaaggacat caaggagtgg
     2341 tacgagatgc tgatgggcca ctgcacctat tttccagagg agctgagaag cgtgaagtac
     2401 gcctataacg ccgatctgta caacgccctg aatgacctga acaacctggt catcaccagg
     2461 gatgagaacg agaagctgga gtactatgag aagttccaga tcatcgagaa cgtgttcaag
     2521 cagaagaaga agcctacact gaagcagatc gccaaggaga tcctggtgaa cgaggaggac
     2581 atcaagggct accgcgtgac ctccacaggc aagccagagt tcaccaatct gaaggtgtat
     2641 cacgatatca aggacatcac agcccggaag gagatcatcg agaacgccga gctgctggat
     2701 cagatcgcca agatcctgac catctatcag agctccgagg acatccagga ggagctgacc
     2761 aacctgaata gcgagctgac acaggaggag atcgagcaga tcagcaatct gaagggctac
     2821 accggcacac acaacctgag cctgaaggcc atcaatctga tcctggatga gctgtggcac
     2881 acaaacgaca atcagatcgc catctttaac cggctgaagc tggtgccaaa gaaggtggac
     2941 ctgtcccagc agaaggagat cccaaccaca ctggtggacg atttcatcct gtctcccgtg
     3001 gtgaagcgga gcttcatcca gagcatcaaa gtgatcaacg ccatcatcaa gaagtacggc
     3061 ctgcccaatg atatcatcat cgagctggcc agggagaaga actccaagga cgcccagaag
     3121 atgatcaatg agatgcagaa gaggaaccgc cagaccaatg agcggatcga ggagatcatc
     3181 agaaccacag gcaaggagaa cgccaagtac ctgatcgaga agatcaagct gcacgatatg
     3241 caggagggca agtgtctgta ttctctggag gccatccctc tggaggacct gctgaacaat
     3301 ccattcaact acgaggtgga tcacatcatc ccccggagcg tgagcttcga caattctttt
     3361 aacaataagg tgctggtgaa gcaggaggag aacagcaaga agggcaatag gacccctttc
     3421 cagtacctgt ctagctccga ttctaagatc agctacgaga cattcaagaa gcacatcctg
     3481 aatctggcca agggcaaggg ccgcatcagc aagaccaaga aggagtacct gctggaggag
     3541 cgggacatca acagattctc cgtgcagaag gacttcatca accggaatct ggtggacacc
     3601 agatacgcca cacgcggcct gatgaatctg ctgcggtctt atttcagagt gaacaatctg
     3661 gatgtgaagg tgaagagcat caacggcggc ttcacctcct ttctgcggag aaagtggaag
     3721 tttaagaagg agcgcaacaa gggctataag caccacgccg aggatgccct gatcatcgcc
     3781 aatgccgact tcatctttaa ggagtggaag aagctggaca aggccaagaa agtgatggag
     3841 aaccagatgt tcgaggagaa gcaggccgag agcatgcccg agatcgagac agagcaggag
     3901 tacaaggaga ttttcatcac acctcaccag atcaagcaca tcaaggactt caaggactac
     3961 aagtattctc acagggtgga taagaagccc aaccgcaagc tgatcaatga caccctgtat
     4021 agcacacgga aggacgataa gggcaatacc ctgatcgtga acaatctgaa cggcctgtac
     4081 gacaaggata atgacaagct gaagaagctg atcaacaagt ctcccgagaa gctgctgatg
     4141 taccaccacg atcctcagac atatcagaag ctgaagctga tcatggagca gtacggcgac
     4201 gagaagaacc cactgtataa gtactatgag gagacaggca actacctgac aaagtatagc
     4261 aagaaggata atggccccgt gatcaagaag atcaagtact atggcaacaa gctgaatgcc
     4321 cacctggaca tcaccgacga ttaccctaac tctcgcaata aggtggtgaa gctgagcctg
     4381 aagccatacc ggttcgacgt gtacctggac aacggcgtgt ataagtttgt gacagtgaag
     4441 aatctggatg tgatcaagaa ggagaactac tatgaggtga acagcaagtg ctacgaggag
     4501 gccaagaagc tgaagaagat cagcaaccag gccgagttca tcgcctcttt ttacaagaat
     4561 gacctgatca agatcaatgg cgagctgtat agagtgatcg gcgtgaacaa tgatctgctg
     4621 aacagaatcg aagtgaatat gatcgacatc acctacaggg agtatctgga gaacatgaat
     4681 gataagaggc cccctcatat catcaagacc atcgcctcta agacacagag catcaagaag
     4741 tacagcacag acatcctggg gaacctgtat gaagtcaaga gcaagaaaca tcctcagatt
     4801 atcaagaaag gctctggcgg ctcaaaaaga accgccgacg gcagcgaatt cgagcccaag
     4861 aagaagagga aagtctaacc ggtcatcatc accatcacca ttgagtttaa acccgctgat
     4921 cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt
     4981 ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat
     5041 cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg
     5101 gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg
     5161 aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc ttggcgtaat
     5221 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     5281 gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa ctcacattaa
     5341 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     5401 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     5461 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     5521 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     5581 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5641 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5701 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5761 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5821 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5881 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5941 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     6001 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6061 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6121 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6181 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6241 ggtctgacac tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6301 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6361 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6421 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6481 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6541 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6601 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6661 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     6721 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     6781 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     6841 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     6901 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6961 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7021 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7081 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7141 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7201 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7261 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7321 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7381 tcgacggatc gggagatcga tctcccgatc ccctagggtc gactctcagt acaatctgct
     7441 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     7501 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     7561 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
     7621 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     7681 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     7741 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     7801 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     7861 aagtgtatc
//
