LOCUS       sequence-441100-        7043 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV genome encoding the N-terminal of haA3A-CBE-G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7043)
  AUTHORS   Yang L, Huo Y, Wang M, Zhang D, Zhang T, Wu H, Rao X, Meng H, Yin S,
            Mei J, Zhang D, Chen X, Lv J, Liu M, Cheng Y, Guan Y, Feng B, Song 
            G, Yi C, Liu M, Zeng F, Wang L, Li D
  TITLE     Engineering APOBEC3A deaminase for highly accurate and efficient 
            base editing.
  JOURNAL   Nat Chem Biol. 2024 Mar 29. doi: 10.1038/s41589-024-01595-4.
  PUBMED    38553609
REFERENCE   2  (bases 1 to 7043)
  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"; journalName: "Nat Chem 
            Biol. 2024 Mar 29. doi: 10.1038/s41589-024-01595-4."
COMMENT     Sequence Label: AAV-haA3A-CBE-G-N
FEATURES             Location/Qualifiers
     source          1..7043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     enhancer        154..533
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        534..737
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     688..708
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     779..798
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        779..797
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             847..867
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1558..3273
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     primer_bind     complement(4027..4044)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4033..4240
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(4273..4297)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     repeat_region   4317..4446
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4508..5096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4588..4607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5267..6058,6059..6127))
                     /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     5890..5909
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6128..6232)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6300..6318
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6356..6378)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6478..6497
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6514..6969)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6651..6672)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6863..6882
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgct agcgacattg attattgact agttattaat
      181 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
      241 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
      301 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
      361 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc
      421 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat
      481 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
      541 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
      601 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg aactttccaa
      661 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg
      721 tctatataag cagagctggt ttagtgaacc gtcagatccg ctagagatcc gcggccgcta
      781 atacgactca ctatagggag agccgccacc atgaaacgga cagccgacgg aagcgagttc
      841 gagtcaccaa agaagaagcg gaaagtcgag gcatctccag caagcggacc aaggcacctg
      901 atggaccccc acatcttcac ctctaacttt aacaatggca tcggcaggca caagacatac
      961 ctgtgctatg aggtggagcg cctggacaac ggcaccagcg tgaagatgga tcagcacaga
     1021 ggcttcctgc acaaccaggc caagaatctg ctgtgcggct tctacggccg gcacgcagag
     1081 ctgagatttc tggacctggt gcctagcctg cagctggatc cagcccagat ctatagggtg
     1141 acctggttca tcagctggtc cccatgcttt tcctggggat gtgcaggaga ggtgcgcgcc
     1201 ttcctgcagg agaatacaca cgtgcggctg agaatctttg ccgcccggat cggcgactat
     1261 gatcctctgt acaaggaggc cctgcagatg ctgagagacg caggagccca ggtgtccatc
     1321 atgacctatg atgagttcaa gcactgctgg gacacatttg tggatcacca gggctgtccc
     1381 tttcagcctt gggacggact ggatgagcac tcccaggccc tgtctggcag gctgagggcc
     1441 atcctgcaga accagggcaa ttctggagga tctagcggag gatcctctgg cagcgagaca
     1501 ccaggaacaa gcgagtcagc aacaccagag agcagtggcg gcagcagcgg cggcagcgac
     1561 aagaagtaca gcatcggcct ggccatcggc accaactctg tgggctgggc cgtgatcacc
     1621 gacgagtaca aggtgcccag caagaaattc aaggtgctgg gcaacaccga ccggcacagc
     1681 atcaagaaga acctgatcgg agccctgctg ttcgacagcg gcgaaacagc cgaggccacc
     1741 cggctgaaga gaaccgccag aagaagatac accagacgga agaaccggat ctgctatctg
     1801 caagagatct tcagcaacga gatggccaag gtggacgaca gcttcttcca cagactggaa
     1861 gagtccttcc tggtggaaga ggataagaag cacgagcggc accccatctt cggcaacatc
     1921 gtggacgagg tggcctacca cgagaagtac cccaccatct accacctgag aaagaaactg
     1981 gtggacagca ccgacaaggc cgacctgcgg ctgatctatc tggccctggc ccacatgatc
     2041 aagttccggg gccacttcct gatcgagggc gacctgaacc ccgacaacag cgacgtggac
     2101 aagctgttca tccagctggt gcagacctac aaccagctgt tcgaggaaaa ccccatcaac
     2161 gccagcggcg tggacgccaa ggccatcctg tctgccagac tgagcaagag cagacggctg
     2221 gaaaatctga tcgcccagct gcccggcgag aagaagaatg gcctgttcgg aaacctgatt
     2281 gccctgagcc tgggcctgac ccccaacttc aagagcaact tcgacctggc cgaggatgcc
     2341 aaactgcagc tgagcaagga cacctacgac gacgacctgg acaacctgct ggcccagatc
     2401 ggcgaccagt acgccgacct gtttctggcc gccaagaacc tgtccgacgc catcctgctg
     2461 agcgacatcc tgagagtgaa caccgagatc accaaggccc ccctgagcgc ctctatgatc
     2521 aagagatacg acgagcacca ccaggacctg accctgctga aagctctcgt gcggcagcag
     2581 ctgcctgaga agtacaaaga gattttcttc gaccagagca agaacggcta cgccggctac
     2641 attgacggcg gagccagcca ggaagagttc tacaagttca tcaagcccat cctggaaaag
     2701 atggacggca ccgaggaact gctcgtgaag ctgaacagag aggacctgct gcggaagcag
     2761 cggaccttcg acaacggcag catcccccac cagatccacc tgggagagct gcacgccatt
     2821 ctgcggcggc aggaagattt ttacccattc ctgaaggaca accgggaaaa gatcgagaag
     2881 atcctgacct tccgcatccc ctactacgtg ggccctctgg ccaggggaaa cagcagattc
     2941 gcctggatga ccagaaagag cgaggaaacc atcaccccct ggaacttcga ggaagtggtg
     3001 gacaagggcg cttccgccca gagcttcatc gagcggatga ccaacttcga taagaacctg
     3061 cccaacgaga aggtgctgcc caagcacagc ctgctgtacg agtacttcac cgtgtataac
     3121 gagctgacca aagtgaaata cgtgaccgag ggaatgagaa agcccgcctt cctgagcggc
     3181 gagcagaaaa aggccatcgt ggacctgctg ttcaagacca accggaaagt gaccgtgaag
     3241 cagctgaaag aggactactt caagaaaatc gagtgcttcg actccgtgga aatctccggc
     3301 gtggaagatc ggttcaacgc ctccctgggc acataccacg atctgctgaa aattatcaag
     3361 gacaaggact tcctggacaa tgaggaaaac gaggacattc tggaagatat cgtgctgacc
     3421 ctgacactgt ttgaggacag agagatgatc gaggaacggc tgaaaaccta tgcccacctg
     3481 ttcgacgaca aagtgatgaa gcagctgaag cggcggagat acaccggctg gggcaggctg
     3541 agccggaagc tgatcaacgg catccgggac aagcagtccg gcaagacaat cctggatttc
     3601 ctgaagtccg acggcttcgc caacagaaac ttcatgcagc tgatccacga cgacagcctg
     3661 acctttaaag aggacatcca gaaagcccag gtgtgcctgt cctacgagac cgaaatcctg
     3721 accgtggagt atgggctgct gcccatcggc aagattgtgg agaagcggat tgaatgcacc
     3781 gtgtatagcg tggacaacaa cggcaacatc tacacccagc ccgtggctca gtggcacgac
     3841 aggggcgagc aggaggtgtt tgagtattgt ctggaggacg gcagcctgat tagagccacc
     3901 aaagaccaca agttcatgac cgtggacggg cagatgctgc ccattgacga gatttttgag
     3961 cgggaacttg acctgatgcg ggtggacaac ctgcccaact aagaattcct agagctcgct
     4021 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     4081 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     4141 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     4201 agggggagga ttgggaagag aatagcaggc atgctgggga ggatccggta ccaaggtcgg
     4261 gcaggaatct agacccagct ttcttgtaca aagttggcat tagagctcgc ggccgcagga
     4321 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4381 gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4441 tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4921 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5041 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5101 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5161 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5221 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5281 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5341 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5401 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5461 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5521 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5581 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5641 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5701 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5761 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5821 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5881 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5941 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6001 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6061 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6121 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6181 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6241 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     6301 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     6361 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     6421 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     6481 catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa
     6541 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca
     6601 aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga
     6661 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     6721 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     6781 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     6841 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg
     6901 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     6961 agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag
     7021 gagaaaatac cgcatcaggc gcc
//
