LOCUS       sequence-270439-        7582 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV genome: expresses the N-terminal of S. aureus v5 AAV-CBE from 
            the Cbh promoter, U6-sgRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7582)
  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 7582)
  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/s41551-019-0501-5"; journalName: "Nat Biomed Eng"; date: 
            "2020-01"; volume: "4"; issue: "1"; pages: "97-110"
COMMENT     Sequence Label: Cbh_v5 AAV-saCBE N-terminal
FEATURES             Location/Qualifiers
     source          1..7582
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(125..142)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(125..141)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(134..156)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      283..738
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(370..389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     580..601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        764..868
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(869..937,938..1729)
                     /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     complement(1087..1106)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1900..2488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2389..2408
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2776..2797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2812..2817,2818..2835,2836..2842)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2850..2866
                     /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     2855..2877
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2874..2890
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2874..2890
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2917..3046
                     /label=AAV2 ITR
     enhancer        3069..3354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        3356..3633
                     /label=chicken beta-actin promoter
     intron          3634..3861
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             3915..3935
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3936..4619
                     /codon_start=1
                     /product="cytidine deaminase (C to U editing enzyme) from 
                     rat"
                     /label=APOBEC-1
                     /note="can use ssDNA as a substrate (Komor et al., 2016)"
                     /translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
                     RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
                     RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
                     AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
                     WATGLK"
     CDS             6663..6683
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6748..6768)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    6944..7168
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(7287..7535)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(7345..7364)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(7515..7535)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   join(7559..7582,1..106)
                     /label=AAV2 ITR
ORIGIN
        1 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
       61 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagcctt aattaaccta
      121 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta
      181 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg
      241 atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc tgtagcggcg
      301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
      361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
      421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
      481 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
      541 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
      601 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
      661 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
      721 tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
      781 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
      841 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
      901 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
      961 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     1021 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     1081 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     1141 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     1201 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     1261 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     1321 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     1381 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     1441 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     1501 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     1561 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     1621 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     1681 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     1741 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     1801 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     1861 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1921 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1981 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     2041 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     2101 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     2161 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     2221 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     2281 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     2341 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     2401 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     2461 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     2521 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     2581 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     2641 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     2701 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     2761 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     2821 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     2881 cagctatgac catgattacg ccagatttaa ttaaggctgc gcgctcgctc gctcactgag
     2941 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
     3001 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg cctctagatc
     3061 agggtacccg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     3121 cgcccattga cgtcaatagt aacgccaata gggactttcc attgacgtca atgggtggag
     3181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     3241 cctattgacg tcaatgacgg taaatggccc gcctggcatt gtgcccagta catgacctta
     3301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtcgag
     3361 gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc cccaattttg
     3421 tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg gggggggcgc
     3481 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
     3541 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
     3601 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
     3661 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     3721 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagct gagcaagagg
     3781 taagggttta agggatggtt ggttggtggg gtattaatgt ttaattacct ggagcacctg
     3841 cctgaaatca ctttttttca ggttggaccg gtgccaccat gaaacggaca gccgacggaa
     3901 gcgagttcga gtcaccaaag aagaagcgga aagtctcctc agagactggg cctgtcgccg
     3961 tcgatccaac cctgcgccgc cggattgaac ctcacgagtt tgaagtgttc tttgaccccc
     4021 gggagctgag aaaggagaca tgcctgctgt acgagatcaa ctggggaggc aggcactcca
     4081 tctggaggca cacctctcag aacacaaata agcacgtgga ggtgaacttc atcgagaagt
     4141 ttaccacaga gcggtacttc tgccccaata ccagatgtag catcacatgg tttctgagct
     4201 ggtccccttg cggagagtgt agcagggcca tcaccgagtt cctgtccaga tatccacacg
     4261 tgacactgtt tatctacatc gccaggctgt atcaccacgc agacccaagg aataggcagg
     4321 gcctgcgcga tctgatcagc tccggcgtga ccatccagat catgacagag caggagtccg
     4381 gctactgctg gcggaacttc gtgaattatt ctcctagcaa cgaggcccac tggcctaggt
     4441 acccacacct gtgggtgcgc ctgtacgtgc tggagctgta ttgcatcatc ctgggcctgc
     4501 ccccttgtct gaatatcctg cggagaaagc agccccagct gaccttcttt acaatcgccc
     4561 tgcagtcttg tcactatcag aggctgccac cccacatcct gtgggccaca ggcctgaagt
     4621 ctggaggatc tagcggagga tcctctggca gcgagacacc aggaacaagc gagtcagcaa
     4681 caccagagag cagtggcggc agcagcggcg gcagcgggaa gcgaaattac attctggggc
     4741 tggccattgg cattacatca gtgggctatg gcatcattga ctacgagaca agggacgtga
     4801 tcgacgccgg cgtgagactg ttcaaggagg ccaacgtgga gaacaatgag ggccggagat
     4861 ccaagagggg agcaaggcgc ctgaagcgga gaaggcgcca cagaatccag agagtgaaga
     4921 agctgctgtt cgattacaac ctgctgaccg accactccga gctgtctggc atcaatcctt
     4981 atgaggccag agtgaagggc ctgtcccaga agctgtctga ggaggagttt agcgccgccc
     5041 tgctgcacct ggcaaagagg agaggcgtgc acaacgtgaa tgaggtggag gaggacaccg
     5101 gcaacgagct gtccacaaag gagcagatca gccgcaattc caaggccctg gaggagaagt
     5161 atgtggccga gctgcagctg gagcggctga agaaggatgg cgaggtgagg ggctccatca
     5221 atcgcttcaa gacctctgac tacgtgaagg aggccaagca gctgctgaag gtgcagaagg
     5281 cctaccacca gctggatcag tcctttatcg atacatatat cgacctgctg gagacaaggc
     5341 gcacatacta tgagggacca ggagagggct ctcccttcgg ctggaaggac atcaaggagt
     5401 ggtacgagat gctgatgggc cactgcacct attttccaga ggagctgaga agcgtgaagt
     5461 acgcctataa cgccgatctg tacaacgccc tgaatgacct gaacaacctg gtcatcacca
     5521 gggatgagaa cgagaagctg gagtactatg agaagttcca gatcatcgag aacgtgttca
     5581 agcagaagaa gaagcctaca ctgaagcaga tcgccaagga gatcctggtg aacgaggagg
     5641 acatcaaggg ctaccgcgtg acctccacag gcaagccaga gttcaccaat ctgaaggtgt
     5701 atcacgatat caaggacatc acagcccgga aggagatcat cgagaacgcc gagctgctgg
     5761 atcagatcgc caagatcctg accatctatc agagctccga ggacatccag gaggagctga
     5821 ccaacctgaa tagcgagctg acacaggagg agatcgagca gatcagcaat ctgaagggct
     5881 acaccggcac acacaacctg agcctgaagg ccatcaatct gatcctggat gagctgtggc
     5941 acacaaacga caatcagatc gccatcttta accggctgaa gctggtgcca aagaaggtgg
     6001 acctgtccca gcagaaggag atcccaacca cactggtgga cgatttcatc ctgtctcccg
     6061 tggtgaagcg gagcttcatc cagagcatca aagtgatcaa cgccatcatc aagaagtacg
     6121 gcctgcccaa tgatatcatc atcgagctgg ccagggagaa gaactccaag gacgcccaga
     6181 agatgatcaa tgagatgcag aagaggaacc gccagaccaa tgagcggatc gaggagatca
     6241 tcagaaccac aggcaaggag aacgccaagt acctgatcga gaagatcaag ctgcacgata
     6301 tgcaggaggg caagtgcctg tcctacgaga cagagatcct gacagtggag tatggcctgc
     6361 tgccaatcgg caagatcgtg gagaagagga tcgagtgtac cgtgtactct gtggataaca
     6421 atggcaacat ctatacacag cccgtggcac agtggcacga taggggagag caggaggtgt
     6481 tcgagtattg cctggaggac ggcagcctga tcagggcaac caaggaccac aagttcatga
     6541 cagtggatgg ccagatgctg cccatcgacg agattttcga gcgggagctg gacctgatga
     6601 gagtggataa cctgcctaat agcggaggca gtaaaagaac agcagacggg agtgagtttg
     6661 agcccaagaa aaagagaaag gtgtaagatc tgataatcaa cctctggatt acaaaatttg
     6721 tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc
     6781 tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta
     6841 taaatcctgg ttagttcttg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     6901 gacaggggct cggctgttgg gcactgacaa ttccgtggtg cgactgtgcc ttctagttgc
     6961 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     7021 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     7081 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     7141 catgctgggg atgcggtggg ctctatggct cgagaaaaaa atctcgccaa caagttgacg
     7201 agataaacac ggcattttgc cttgttttag tagattctgt aattttcatt acagagtact
     7261 aaaaccgaga cgcattaatc gtctcgggtg tttcgtcctt tccacaagat atataaagcc
     7321 aagaaatcga aatactttca agttacggta agcatatgat agtccatttt aaaacataat
     7381 tttaaaactg caaactaccc aagaaattat tactttctac gtcacgtatt ttgtactaat
     7441 atctttgtgt ttacagtcaa attaattcta attatctctc taacagcctt gtatcgtata
     7501 tgcaaatatg aaggaatcat gggaaatagg ccctcttcct gcccgacctt gcggccgcag
     7561 gaacccctag tgatggagtt gg
//
