A013-CMV-1115Nterm

LOCUS       A013-CMV-1115Nterm      7691 bp DNA     circular SYN 31-MAR-2026
DEFINITION  AAV genome: expresses the N-terminal of v5 AAV-ABE from the Cbh 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7691)
  AUTHORS   Levy JM, Yeh WH, Pendse N, Davis JR, Hennessey E, Butcher R, Koblan 
            LW, Comander J, Liu Q, Liu DR
  TITLE     Cytosine and adenine base editing of the brain, liver, retina, heart
            and skeletal muscle of mice via adeno-associated viruses.
  JOURNAL   Nat Biomed Eng. 2020 Jan;4(1):97-110. doi: 
            10.1038/s41551-019-0501-5. Epub 2020 Jan 14.
  PUBMED    31937940
REFERENCE   2  (bases 1 to 7691)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 31, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41551-019-0501-5"; journalName: "Nat Biomed Eng"; date: 
            "2020-01"; volume: "4"; issue: "1"; pages: "97-110"
FEATURES             Location/Qualifiers
     source          1..7691
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        153..438
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        439..642
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     593..613
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      661..666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             670..723
                     /codon_start=1
                     /label=NLS(BE4Max1)
                     /translation="KRTADGSEFESPKKKRKV"
     CDS             724..4065
                     /codon_start=1
                     /label=Cas9-N(2-1115)
                     /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRN"
     CDS             4066..4371
                     /codon_start=1
                     /label=RmaDnaBIntN
                     /translation="CLAGDTLITLADGRRVPIRELVSQQNFSVWALNPQTYRLERARVS
                     RAFCTGIKPVYRLTTRLGRSIRATANHRFLTPQGWKRVDELQPGDYLALPRRIPTAS"
     terminator      4380..4540
                     /label=WPRE_Gamma
                     /note="/created_by=User"
                     /note="/modified_by=User"
     misc_feature    4629..4743
                     /label=sv40SL
     repeat_region   4752..4881
                     /label=AAV2 ITR
     primer_bind     complement(4900..4917)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4900..4916)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4909..4931)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5058..5513
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5539..5643
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5644..6504
                     /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"
     misc_feature    complement(6557..6562)
                     /label=PolyAMotif2
     rep_origin      6675..7263
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    7551..7572
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7587..7617
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7625..7641
                     /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)."
     primer_bind     7630..7652
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7649..7665
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7649..7665
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    complement(7681..7686)
                     /label=PolyAMotif2
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcctcta gatcagggta cccgttacat aacttacggt aaatggcccg
      181 cctggctgac cgcccaacga cccccgccca ttgacgtcaa tagtaacgcc aatagggact
      241 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      301 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      361 cattgtgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      421 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      481 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      541 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      601 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactaccggt
      661 gccaccatga aacggacagc cgacggaagc gagttcgagt caccaaagaa gaagcggaaa
      721 gtcgacaaga agtacagcat cggcctggac atcggcacca actctgtggg ctgggccgtg
      781 atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa caccgaccgg
      841 cacagcatca agaagaacct gatcggagcc ctgctgttcg acagcggcga aacagccgag
      901 gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa ccggatctgc
      961 tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt cttccacaga
     1021 ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc catcttcggc
     1081 aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca cctgagaaag
     1141 aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc cctggcccac
     1201 atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga caacagcgac
     1261 gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga ggaaaacccc
     1321 atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag caagagcaga
     1381 cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct gttcggaaac
     1441 ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga cctggccgag
     1501 gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa cctgctggcc
     1561 cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc cgacgccatc
     1621 ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct gagcgcctct
     1681 atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg
     1741 cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa cggctacgcc
     1801 ggctacattg acggcggagc cagccaggaa gagttctaca agttcatcaa gcccatcctg
     1861 gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga cctgctgcgg
     1921 aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg agagctgcac
     1981 gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg ggaaaagatc
     2041 gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag gggaaacagc
     2101 agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa cttcgaggaa
     2161 gtggtggaca agggcgcttc cgcccagagc ttcatcgagc ggatgaccaa cttcgataag
     2221 aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta cttcaccgtg
     2281 tataacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc cgccttcctg
     2341 agcggcgagc agaaaaaggc catcgtggac ctgctgttca agaccaaccg gaaagtgacc
     2401 gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc
     2461 tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct gctgaaaatt
     2521 atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga agatatcgtg
     2581 ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa aacctatgcc
     2641 cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac cggctggggc
     2701 aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa gacaatcctg
     2761 gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat ccacgacgac
     2821 agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg cgatagcctg
     2881 cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat cctgcagaca
     2941 gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga gaacatcgtg
     3001 atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag ccgcgagaga
     3061 atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa agaacacccc
     3121 gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca gaatgggcgg
     3181 gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga tgtggacgct
     3241 atcgtgcctc agagctttct gaaggacgac tccatcgaca acaaggtgct gaccagaagc
     3301 gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag
     3361 aactactggc ggcagctgct gaacgccaag ctgattaccc agagaaagtt cgacaatctg
     3421 accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat caagagacag
     3481 ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc ccggatgaac
     3541 actaagtacg acgagaatga caagctgatc cgggaagtga aagtgatcac cctgaagtcc
     3601 aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga gatcaacaac
     3661 taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag
     3721 taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag
     3781 atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt cttctacagc
     3841 aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat ccggaagcgg
     3901 cctctgatcg agacaaacgg cgaaaccggg gagatcgtgt gggataaggg ccgggatttt
     3961 gccaccgtgc ggaaagtgct gagcatgccc caagtgaata tcgtgaaaaa gaccgaggtg
     4021 cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaactgtct ggctggcgat
     4081 actctcatta ccctggccga tggacgacga gtgcctatta gagaactggt gtcacagcag
     4141 aatttttccg tgtgggctct gaatcctcag acttaccgcc tggagagggc tagagtgagt
     4201 agagctttct gtaccggcat caaacctgtg taccgcctca ccactagact ggggagatcc
     4261 attagggcca ctgccaacca ccgatttctc acacctcagg gctggaaacg agtcgatgaa
     4321 ctccagcctg gagattacct ggctctgcct aggagaatcc ctactgcctc ctaagatctg
     4381 ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
     4441 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
     4501 gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc acggcggaac
     4561 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     4621 ccgtggtgtt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc atctagcttt
     4681 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     4741 gttgcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     4801 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     4861 agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg ttttacaacg
     4921 tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt
     4981 cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag
     5041 cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     5101 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     5161 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5221 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     5281 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5341 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     5401 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     5461 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     5521 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5581 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5641 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5701 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5761 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5821 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5881 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5941 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     6001 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     6061 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     6121 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     6181 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     6241 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     6301 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     6361 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     6421 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     6481 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6541 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6601 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6661 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6721 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6781 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6841 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6901 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6961 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     7021 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     7081 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     7141 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     7201 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     7261 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     7321 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     7381 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     7441 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     7501 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7561 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7621 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaga
     7681 tttaattaag g
//