LOCUS       sequence-272939-        8388 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 8388)
  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 8388)
  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: LbABE8e-dimer
FEATURES             Location/Qualifiers
     source          1..8388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        19..398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        399..602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     553..573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     644..663
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        644..662
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             712..732
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1921..5598
                     /codon_start=1
                     /product="CRISPR-associated protein from Lachnospiraceae 
                     bacterium ND2006 (Zetsche et al., 2015)"
                     /label=LbCpf1
                     /translation="SKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAED
                     YKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKEI
                     AKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFSE
                     EAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEGE
                     FFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVLS
                     DRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKNG
                     PAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQL
                     QEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKDL
                     LDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYSK
                     DKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNGN
                     YEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKLI
                     DFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLVE
                     EGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKKE
                     ELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKIN
                     TEVRVLLKHDDNPYVIGIARGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTDY
                     HSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSGF
                     KNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQN
                     GFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALDY
                     KNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQQ
                     GDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRNY
                     EAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTSV
                     K"
     CDS             5641..5661
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5670..5687
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5710..5727)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5716..5940
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6011..6027)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6011..6027)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6024..6046)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6035..6051
                     /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(6059..6065,6066..6083,6084..6089))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6104..6125
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6242..6259)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6413..7001)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6493..6512)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7172..7963,7964..8032))
                     /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     7795..7814
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8033..8137)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8216..8235)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
       61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      601 ctggtttagt gaaccgtcag atccgctaga gatccgcggc cgctaatacg actcactata
      661 gggagagccg ccaccatgaa acggacagcc gacggaagcg agttcgagtc accaaagaag
      721 aagcggaaag tctctgaagt cgagtttagc cacgagtatt ggatgaggca cgcactgacc
      781 ctggcaaagc gagcatggga tgaaagagaa gtccccgtgg gcgccgtgct ggtgcacaac
      841 aatagagtga tcggagaggg atggaacagg ccaatcggcc gccacgaccc taccgcacac
      901 gcagagatca tggcactgag gcagggaggc ctggtcatgc agaattaccg cctgatcgat
      961 gccaccctgt atgtgacact ggagccatgc gtgatgtgcg caggagcaat gatccacagc
     1021 aggatcggaa gagtggtgtt cggagcacgg gacgccaaga ccggcgcagc aggctccctg
     1081 atggatgtgc tgcaccaccc cggcatgaac caccgggtgg agatcacaga gggaatcctg
     1141 gcagacgagt gcgccgccct gctgagcgat ttctttagaa tgcggagaca ggagatcaag
     1201 gcccagaaga aggcacagag ctccaccgac tctggaggat ctagcggagg atcctctgga
     1261 agcgagacac caggcacaag cgagtccgcc acaccagaga gctccggcgg ctcctccgga
     1321 ggatcctctg aggtggagtt ttcccacgag tactggatga gacatgccct gaccctggcc
     1381 aagagggcac gggatgagag ggaggtgcct gtgggagccg tgctggtgct gaacaataga
     1441 gtgatcggcg agggctggaa cagagccatc ggcctgcacg acccaacagc ccatgccgaa
     1501 attatggccc tgagacaggg cggcctggtc atgcagaact acagactgat tgacgccacc
     1561 ctgtacgtga cattcgagcc ttgcgtgatg tgcgccggcg ccatgatcca ctctaggatc
     1621 ggccgcgtgg tgtttggcgt gaggaactca aaaagaggcg ccgcaggctc cctgatgaac
     1681 gtgctgaact accccggcat gaatcaccgc gtcgaaatta ccgagggaat cctggcagat
     1741 gaatgtgccg ccctgctgtg cgatttctat cggatgccta gacaggtgtt caatgctcag
     1801 aagaaggccc agagctccat caactccgga ggatctagcg gaggatcctc tggcagcgag
     1861 acaccaggaa caagcgagtc agcaacacca gagagcagtg gcggcagcag cggcggcagc
     1921 agcaagctgg agaagtttac aaactgctac tccctgtcta agaccctgag gttcaaggcc
     1981 atccctgtgg gcaagaccca ggagaacatc gacaataagc ggctgctggt ggaggacgag
     2041 aagagagccg aggattataa gggcgtgaag aagctgctgg atcgctacta tctgtctttt
     2101 atcaacgacg tgctgcacag catcaagctg aagaatctga acaattacat cagcctgttc
     2161 cggaagaaaa ccagaaccga gaaggagaat aaggagctgg agaacctgga gatcaatctg
     2221 cggaaggaga tcgccaaggc cttcaagggc aacgagggct acaagtccct gtttaagaag
     2281 gatatcatcg agacaatcct gccagagttc ctggacgata aggacgagat cgccctggtg
     2341 aacagcttca atggctttac cacagccttc accggcttct ttgataacag agagaatatg
     2401 ttttccgagg aggccaagag cacatccatc gccttcaggt gtatcaacga gaatctgacc
     2461 cgctacatct ctaatatgga catcttcgag aaggtggacg ccatctttga taagcacgag
     2521 gtgcaggaga tcaaggagaa gatcctgaac agcgactatg atgtggagga tttctttgag
     2581 ggcgagttct ttaactttgt gctgacacag gagggcatcg acgtgtataa cgccatcatc
     2641 ggcggcttcg tgaccgagag cggcgagaag atcaagggcc tgaacgagta catcaacctg
     2701 tataatcaga aaaccaagca gaagctgcct aagtttaagc cactgtataa gcaggtgctg
     2761 agcgatcggg agtctctgag cttctacggc gagggctata catccgatga ggaggtgctg
     2821 gaggtgttta gaaacaccct gaacaagaac agcgagatct tcagctccat caagaagctg
     2881 gagaagctgt tcaagaattt tgacgagtac tctagcgccg gcatctttgt gaagaacggc
     2941 cccgccatca gcacaatctc caaggatatc ttcggcgagt ggaacgtgat ccgggacaag
     3001 tggaatgccg agtatgacga tatccacctg aagaagaagg ccgtggtgac cgagaagtac
     3061 gaggacgatc ggagaaagtc cttcaagaag atcggctcct tttctctgga gcagctgcag
     3121 gagtacgccg acgccgatct gtctgtggtg gagaagctga aggagatcat catccagaag
     3181 gtggatgaga tctacaaggt gtatggctcc tctgagaagc tgttcgacgc cgattttgtg
     3241 ctggagaaga gcctgaagaa gaacgacgcc gtggtggcca tcatgaagga cctgctggat
     3301 tctgtgaaga gcttcgagaa ttacatcaag gccttctttg gcgagggcaa ggagacaaac
     3361 agggacgagt ccttctatgg cgattttgtg ctggcctacg acatcctgct gaaggtggac
     3421 cacatctacg atgccatccg caattatgtg acccagaagc cctactctaa ggataagttc
     3481 aagctgtatt ttcagaaccc tcagttcatg ggcggctggg acaaggataa ggagacagac
     3541 tatcgggcca ccatcctgag atacggctcc aagtactatc tggccatcat ggataagaag
     3601 tacgccaagt gcctgcagaa gatcgacaag gacgatgtga acggcaatta cgagaagatc
     3661 aactataagc tgctgcccgg ccctaataag atgctgccaa aggtgttctt ttctaagaag
     3721 tggatggcct actataaccc cagcgaggac atccagaaga tctacaagaa tggcacattc
     3781 aagaagggcg atatgtttaa cctgaatgac tgtcacaagc tgatcgactt ctttaaggat
     3841 agcatctccc ggtatccaaa gtggtccaat gcctacgatt tcaacttttc tgagacagag
     3901 aagtataagg acatcgccgg cttttacaga gaggtggagg agcagggcta taaggtgagc
     3961 ttcgagtctg ccagcaagaa ggaggtggat aagctggtgg aggagggcaa gctgtatatg
     4021 ttccagatct ataacaagga cttttccgat aagtctcacg gcacacccaa tctgcacacc
     4081 atgtacttca agctgctgtt tgacgagaac aatcacggac agatcaggct gagcggagga
     4141 gcagagctgt tcatgaggcg cgcctccctg aagaaggagg agctggtggt gcacccagcc
     4201 aactccccta tcgccaacaa gaatccagat aatcccaaga aaaccacaac cctgtcctac
     4261 gacgtgtata aggataagag gttttctgag gaccagtacg agctgcacat cccaatcgcc
     4321 atcaataagt gccccaagaa catcttcaag atcaatacag aggtgcgcgt gctgctgaag
     4381 cacgacgata acccctatgt gatcggcatc gccaggggtg agcgcaatct gctgtatatc
     4441 gtggtggtgg acggcaaggg caacatcgtg gagcagtatt ccctgaacga gatcatcaac
     4501 aacttcaacg gcatcaggat caagacagat taccactctc tgctggacaa gaaggagaag
     4561 gagaggttcg aggcccgcca gaactggacc tccatcgaga atatcaagga gctgaaggcc
     4621 ggctatatct ctcaggtggt gcacaagatc tgcgagctgg tggagaagta cgatgccgtg
     4681 atcgccctgg aggacctgaa ctctggcttt aagaatagcc gcgtgaaggt ggagaagcag
     4741 gtgtatcaga agttcgagaa gatgctgatc gataagctga actacatggt ggacaagaag
     4801 tctaatcctt gtgcaacagg cggcgccctg aagggctatc agatcaccaa taagttcgag
     4861 agctttaagt ccatgtctac ccagaacggc ttcatctttt acatccctgc ctggctgaca
     4921 tccaagatcg atccatctac cggctttgtg aacctgctga aaaccaagta taccagcatc
     4981 gccgattcca agaagttcat cagctccttt gacaggatca tgtacgtgcc cgaggaggat
     5041 ctgttcgagt ttgccctgga ctataagaac ttctctcgca cagacgccga ttacatcaag
     5101 aagtggaagc tgtactccta cggcaaccgg atcagaatct tccggaatcc taagaagaac
     5161 aacgtgttcg actgggagga ggtgtgcctg accagcgcct ataaggagct gttcaacaag
     5221 tacggcatca attatcagca gggcgatatc agagccctgc tgtgcgagca gtccgacaag
     5281 gccttctact ctagctttat ggccctgatg agcctgatgc tgcagatgcg gaacagcatc
     5341 acaggccgca ccgacgtgga ttttctgatc agccctgtga agaactccga cggcatcttc
     5401 tacgatagcc ggaactatga ggcccaggag aatgccatcc tgccaaagaa cgccgacgcc
     5461 aatggcgcct ataacatcgc cagaaaggtg ctgtgggcca tcggccagtt caagaaggcc
     5521 gaggacgaga agctggataa ggtgaagatc gccatctcta acaaggagtg gctggagtac
     5581 gcccagacca gcgtgaagtc tggcggctca aaaagaaccg ccgacggcag cgaattcgag
     5641 cccaagaaga agaggaaagt ctaaccggtc atcatcacca tcaccattga gtttaaaccc
     5701 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     5761 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     5821 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     5881 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     5941 cttctgaggc ggaaagaacc agctggggct cgataccgtc gacctctagc tagagcttgg
     6001 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6061 acatacgagc cggaagcata aagtgtaaag cctagggtgc ctaatgagtg agctaactca
     6121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6241 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6301 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6361 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6421 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6481 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6541 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6601 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6661 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6721 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6781 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6841 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6901 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     6961 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7021 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7081 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7141 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7201 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7261 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aattataccg cgagacccac
     7321 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7381 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7441 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7501 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7561 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7621 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7681 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7741 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7801 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7861 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7921 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     7981 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8041 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8101 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8161 ctgacgtcga cggatcggga gatcgatctc ccgatcccat agggtcgact ctcagtacaa
     8221 tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg
     8281 ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca
     8341 tgaagaatct gcttagggtt aggcgttttg cgctgcttcg cgatgtac
//
