LOCUS       sequence-441098-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV genome encoding the C-terminal of haA3A-CBE-G, and sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  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 7126)
  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-C
FEATURES             Location/Qualifiers
     source          1..7126
                     /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
     promoter        154..402
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     154..174
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     325..344
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        421..496
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        516..895
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        896..1099
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1050..1070
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3252..3272
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3282..3338
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             3339..3587
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             3600..3620
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3630..3683
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3684..3932
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             3975..3995
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4002..4082
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
     primer_bind     complement(4110..4127)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4116..4323
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(4356..4380)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     repeat_region   4400..4529
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(4591..5179)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4671..4690)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5350..6141,6142..6210))
                     /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     5973..5992
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6211..6315)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6383..6401
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6439..6461)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6561..6580
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6597..7052)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6734..6755)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6946..6965
                     /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 agcgagggcc tatttcccat gattccttca
      181 tatttgcata tacgatacaa ggctgttaga gagataattg gaattaattt gactgtaaac
      241 acaaagatat tagtacaaaa tacgtgacgt agaaagtaat aatttcttgg gtagtttgca
      301 gttttaaaat tatgttttaa aatggactat catatgctta ccgtaacttg aaagtatttc
      361 gatttcttgg ctttatatat cttgtggaaa ggacgaaaca ccgggtcttc gagaagacct
      421 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      481 ggcaccgagt cggtgctttt tttatgcatg gtaccgacat tgattattga ctagttatta
      541 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      601 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      661 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      721 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      781 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      841 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      901 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      961 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     1021 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
     1081 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagaacc ggtgccacca
     1141 tgataaagat tgccaccaga aagtatctgg gcaagcagaa cgtgtatgac atcggcgtgg
     1201 agagagacca caacttcgcc ctgaagaacg gcttcatcgc cagctccggc cagggcgata
     1261 gcctgcacga gcacattgcc aatctggccg gcagccccgc cattaagaag ggcatcctgc
     1321 agacagtgaa ggtggtggac gagctcgtga aagtgatggg ccggcacaag cccgagaaca
     1381 tcgtgatcga aatggccaga gagaaccaga ccacccagaa gggacagaag aacagccgcg
     1441 agagaatgaa gcggatcgaa gagggcatca aagagctggg cagccagatc ctgaaagaac
     1501 accccgtgga aaacacccag ctgcagaacg agaagctgta cctgtactac ctgcagaatg
     1561 ggcgggatat gtacgtggac caggaactgg acatcaaccg gctgtccgac tacgatgtgg
     1621 accatatcgt gcctcagagc tttctgaagg acgactccat cgacaacaag gtgctgacca
     1681 gaagcgacaa gaaccggggc aagagcgaca acgtgccctc cgaagaggtc gtgaagaaga
     1741 tgaagaacta ctggcggcag ctgctgaacg ccaagctgat tacccagaga aagttcgaca
     1801 atctgaccaa ggccgagaga ggcggcctga gcgaactgga taaggccggc ttcatcaaga
     1861 gacagctggt ggaaacccgg cagatcacaa agcacgtggc acagatcctg gactcccgga
     1921 tgaacactaa gtacgacgag aatgacaagc tgatccggga agtgaaagtg atcaccctga
     1981 agtccaagct ggtgtccgat ttccggaagg atttccagtt ttacaaagtg cgcgagatca
     2041 acaactacca ccacgcccac gacgcctacc tgaacgccgt cgtgggaacc gccctgatca
     2101 aaaagtaccc taagctggaa agcgagttcg tgtacggcga ctacaaggtg tacgacgtgc
     2161 ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc taccgccaag tacttcttct
     2221 acagcaacat catgaacttt ttcaagaccg agattaccct ggccaacggc gagatccgga
     2281 agcggcctct gatcgagaca aacggcgaaa ccggggagat cgtgtgggat aagggccggg
     2341 attttgccac cgtgcggaaa gtgctgagca tgccccaagt gaatatcgtg aaaaagaccg
     2401 aggtgcagac aggcggcttc agcaaagagt ctatcctgcc caagaggaac agcgataagc
     2461 tgatcgccag aaagaaggac tgggacccta agaagtacgg cggcttcgac agccccaccg
     2521 tggcctattc tgtgctggtg gtggccaaag tggaaaaggg caagtccaag aaactgaaga
     2581 gtgtgaaaga gctgctgggg atcaccatca tggaaagaag cagcttcgag aagaatccca
     2641 tcgactttct ggaagccaag ggctacaaag aagtgaaaaa ggacctgatc atcaagctgc
     2701 ctaagtactc cctgttcgag ctggaaaacg gccggaagag aatgctggcc tctgccggcg
     2761 aactgcagaa gggaaacgaa ctggccctgc cctccaaata tgtgaacttc ctgtacctgg
     2821 ccagccacta tgagaagctg aagggctccc ccgaggataa tgagcagaaa cagctgtttg
     2881 tggaacagca caagcactac ctggacgaga tcatcgagca gatcagcgag ttctccaaga
     2941 gagtgatcct ggccgacgct aatctggaca aagtgctgtc cgcctacaac aagcaccggg
     3001 ataagcccat cagagagcag gccgagaata tcatccacct gtttaccctg accaatctgg
     3061 gagcccctgc cgccttcaag tactttgaca ccaccatcga ccggaagagg tacaccagca
     3121 ccaaagaggt gctggacgcc accctgatcc accagagcat caccggcctg tacgagacac
     3181 ggatcgacct gtctcagctg ggaggtgact ctggcggctc aaaaagaacc gccgacggca
     3241 gcgaattcga gcccaagaag aagaggaaag tcggaagcgg agctactaac ttcagcctgc
     3301 tgaagcaggc tggagacgtg gaggagaacc ctggacctac taatctgagc gacatcattg
     3361 agaaggagac tgggaaacag ctggtcattc aggagtccat cctgatgctg cctgaggagg
     3421 tggaggaagt gatcggcaac aagccagagt ctgacatcct ggtgcacacc gcctacgacg
     3481 agtccacaga tgagaatgtg atgctgctga cctctgacgc ccccgagtat aagccttggg
     3541 ccctggtcat ccaggattct aacggcgaga ataagatcaa gatgctgagc ggaggatccc
     3601 ccaagaagaa gaggaaagtc ggatccggag agggcagagg aagtctgcta acatgcggtg
     3661 acgtcgagga gaatcctggc ccaaccaacc tgtctgacat catcgagaag gagacaggca
     3721 agcagctggt catccaggag agcatcctga tgctgcccga agaagtcgaa gaagtgatcg
     3781 gaaacaagcc tgagagcgat atcctggtcc ataccgccta cgacgagagt accgacgaaa
     3841 atgtgatgct gctgacatcc gacgccccag agtataagcc ctgggctctg gtcatccagg
     3901 attccaacgg agagaacaaa atcaaaatgc tgtctggcgg ctcaaaaaga accgccgacg
     3961 gcagcgaatt cgagcccaag aagaagagga aagtcggatc ctacccatac gatgttccag
     4021 attacgctta tccctacgac gtgcctgatt atgcataccc atatgatgtc cccgactatg
     4081 cctaagaatt cctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     4141 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     4201 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4261 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg
     4321 ggaggatccg gtaccaaggt cgggcaggaa tctagaccca gctttcttgt acaaagttgg
     4381 cattagagct cgcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4441 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg cggcctcagt
     4501 gagcgagcga gcgcgcagct gcctgcagga catgtgagca aaaggccagc aaaaggccag
     4561 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4621 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4681 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4741 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4801 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4861 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4921 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4981 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5041 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5101 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5161 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5221 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5281 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5341 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     5401 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     5461 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     5521 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     5581 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     5641 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     5701 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     5761 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     5821 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     5881 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     5941 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     6001 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     6061 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     6121 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     6181 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     6241 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     6301 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat
     6361 tatcatgaca ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt
     6421 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     6481 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     6541 tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc accataaaat
     6601 tgtaaacgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     6661 taaccaatag accgaaatcg gcaaaatccc ttataaatca aaagaatagc ccgagataga
     6721 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     6781 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat cacccaaatc
     6841 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     6901 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     6961 aggagcgggc gctaaggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     7021 cgccgcgctt aatgcgccgc tacagggcgc gtactatggt tgctttgacg tatgcggtgt
     7081 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgcc
//
