LOCUS       sequence-265153-        6843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV genome: expresses the C-terminal of S. aureus v5 AAV-CBE, and 
            U6-sgRNA (KKH variant)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6843)
  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 6843)
  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_KKH C-terminal
FEATURES             Location/Qualifiers
     source          1..6843
                     /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        765..869
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(870..938,939..1730)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVVMVTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1088..1107)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1901..2489
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2390..2409
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    2777..2798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2813..2818,2819..2836,2837..2843)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2851..2867
                     /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     2856..2878
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2875..2891
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2875..2891
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   2918..3047
                     /label=AAV2 ITR
     enhancer        3070..3355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer; 
                     contains an 18-bp deletion relative to the standard CMV 
                     enhancer"
     promoter        3357..3634
                     /label=chicken beta-actin promoter
     intron          3635..3863
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             3917..3937
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5633..5881
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5924..5944
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6009..6029)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    6205..6429
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(6548..6796)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6606..6625)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6776..6796)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   join(6820..6843,1..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 tattaacgtt tataatttca ggtggcatct ttcggggaaa tgtgcgcgga acccctattt
      781 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
      841 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
      901 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
      961 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     1021 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     1081 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     1141 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     1201 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     1261 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     1321 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     1381 taccaaacga cgagcgtgac accacgatgc ctgtagtaat ggtaacaacg ttgcgcaaac
     1441 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     1501 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     1561 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     1621 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     1681 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     1741 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1801 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1861 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1921 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1981 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     2041 atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2101 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2161 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2221 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2281 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2341 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2401 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2461 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2521 tggccttttg ctgcggtttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2581 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     2641 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     2701 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     2761 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     2821 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
     2881 acagctatga ccatgattac gccagattta attaaggctg cgcgctcgct cgctcactga
     2941 ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     3001 gcgagcgcgc agagagggag tggccaactc catcactagg ggttcctgcg gcctctagat
     3061 cagggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     3121 ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc aatgggtgga
     3181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     3241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt acatgacctt
     3301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga
     3361 ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt
     3421 gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggggcg
     3481 cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga gaggtgcggc
     3541 ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc ggcggcggcg
     3601 gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc tgccttcgcc
     3661 ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg accgcgttac
     3721 tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag ctgagcaaga
     3781 ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac ctggagcacc
     3841 tgcctgaaat cacttttttt caggttggac cggtgccacc atgaaacgga cagccgacgg
     3901 aagcgagttc gagtcaccaa agaagaagcg gaaagtcatc aagattgcta cacggaaata
     3961 cctgggaaag cagaacgtgt acgacatcgg cgtggagcgg gatcacaact tcgccctgaa
     4021 gaatggcttt atcgccagca attgtctgta ttctctggag gccatccctc tggaggacct
     4081 gctgaacaat ccattcaact acgaggtgga tcacatcatc ccccggagcg tgagcttcga
     4141 caattctttt aacaataagg tgctggtgaa gcaggaggag aacagcaaga agggcaatag
     4201 gacccctttc cagtacctgt ctagctccga ttctaagatc agctacgaga cattcaagaa
     4261 gcacatcctg aatctggcca agggcaaggg ccgcatcagc aagaccaaga aggagtacct
     4321 gctggaggag cgggacatca acagattctc cgtgcagaag gacttcatca accggaatct
     4381 ggtggacacc agatacgcca cacgcggcct gatgaatctg ctgcggtctt atttcagagt
     4441 gaacaatctg gatgtgaagg tgaagagcat caacggcggc ttcacctcct ttctgcggag
     4501 aaagtggaag tttaagaagg agcgcaacaa gggctataag caccacgccg aggatgccct
     4561 gatcatcgcc aatgccgact tcatctttaa ggagtggaag aagctggaca aggccaagaa
     4621 agtgatggag aaccagatgt tcgaggagaa gcaggccgag agcatgcccg agatcgagac
     4681 agagcaggag tacaaggaga ttttcatcac acctcaccag atcaagcaca tcaaggactt
     4741 caaggactac aagtattctc acagggtgga taagaagccc aaccgcaagc tgatcaatga
     4801 caccctgtat agcacacgga aggacgataa gggcaatacc ctgatcgtga acaatctgaa
     4861 cggcctgtac gacaaggata atgacaagct gaagaagctg atcaacaagt ctcccgagaa
     4921 gctgctgatg taccaccacg atcctcagac atatcagaag ctgaagctga tcatggagca
     4981 gtacggcgac gagaagaacc cactgtataa gtactatgag gagacaggca actacctgac
     5041 aaagtatagc aagaaggata atggccccgt gatcaagaag atcaagtact atggcaacaa
     5101 gctgaatgcc cacctggaca tcaccgacga ttaccctaac tctcgcaata aggtggtgaa
     5161 gctgagcctg aagccatacc ggttcgacgt gtacctggac aacggcgtgt ataagtttgt
     5221 gacagtgaag aatctggatg tgatcaagaa ggagaactac tatgaggtga acagcaagtg
     5281 ctacgaggag gccaagaagc tgaagaagat cagcaaccag gccgagttca tcgcctcttt
     5341 ttacaagaat gacctgatca agatcaatgg cgagctgtat agagtgatcg gcgtgaacaa
     5401 tgatctgctg aacagaatcg aagtgaatat gatcgacatc acctacaggg agtatctgga
     5461 gaacatgaat gataagaggc cccctcatat catcaagacc atcgcctcta agacacagag
     5521 catcaagaag tacagcacag acatcctggg gaacctgtat gaagtcaaga gcaagaaaca
     5581 tcctcagatt atcaagaaag gcagcggcgg gagcggcggg agcgggggga gcactaatct
     5641 gagcgacatc attgagaagg agactgggaa acagctggtc attcaggagt ccatcctgat
     5701 gctgcctgag gaggtggagg aagtgatcgg caacaagcca gagtctgaca tcctggtgca
     5761 caccgcctac gacgagtcca cagatgagaa tgtgatgctg ctgacctctg acgcccccga
     5821 gtataagcct tgggccctgg tcatccagga ttctaacggc gagaataaga tcaagatgct
     5881 gagcggagga tccaaaagaa ccgccgacgg cagcgaattc gagcccaaga agaagaggaa
     5941 agtctaagat ctgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     6001 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     6061 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttagttctt
     6121 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     6181 ggcactgaca attccgtggt gcgactgtgc cttctagttg ccagccatct gttgtttgcc
     6241 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa
     6301 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg
     6361 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg
     6421 gctctatggc tcgagaaaaa aatctcgcca acaagttgac gagataaaca cggcattttg
     6481 ccttgtttta gtagattctg taattttcat tacagagtac taaaaccgag acgcattaat
     6541 cgtctcgggt gtttcgtcct ttccacaaga tatataaagc caagaaatcg aaatactttc
     6601 aagttacggt aagcatatga tagtccattt taaaacataa ttttaaaact gcaaactacc
     6661 caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg tttacagtca
     6721 aattaattct aattatctct ctaacagcct tgtatcgtat atgcaaatat gaaggaatca
     6781 tgggaaatag gccctcttcc tgcccgacct tgcggccgca ggaaccccta gtgatggagt
     6841 tgg
//
