LOCUS       sequence-270521-        6840 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 6840)
  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 6840)
  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..6840
                     /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..3632
                     /label=chicken beta-actin promoter
     intron          3633..3860
                     /label=hybrid intron
                     /note="hybrid between chicken beta-actin (CBA) and minute 
                     virus of mice (MMV) introns (Gray et al., 2011)"
     CDS             3914..3934
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5630..5878
                     /codon_start=1
                     /product="uracil-DNA glycosylase inhibitor from a Bacillus 
                     subtilis bacteriophage (Mol et al., 1995)"
                     /label=UGI
                     /translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
                     YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
     CDS             5921..5941
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(6006..6026)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     polyA_signal    6202..6426
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(6545..6793)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(6603..6622)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(6773..6793)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   join(6817..6840,1..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 ggggggcgcg
     3481 cgccaggcgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg
     3541 cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc
     3601 ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg
     3661 tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc
     3721 cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt
     3781 aagggtttaa gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc
     3841 ctgaaatcac tttttttcag gttggaccgg tgccaccatg aaacggacag ccgacggaag
     3901 cgagttcgag tcaccaaaga agaagcggaa agtcatcaag attgctacac ggaaatacct
     3961 gggaaagcag aacgtgtacg acatcggcgt ggagcgggat cacaacttcg ccctgaagaa
     4021 tggctttatc gccagcaatt gtctgtattc tctggaggcc atccctctgg aggacctgct
     4081 gaacaatcca ttcaactacg aggtggatca catcatcccc cggagcgtga gcttcgacaa
     4141 ttcttttaac aataaggtgc tggtgaagca ggaggagaac agcaagaagg gcaataggac
     4201 ccctttccag tacctgtcta gctccgattc taagatcagc tacgagacat tcaagaagca
     4261 catcctgaat ctggccaagg gcaagggccg catcagcaag accaagaagg agtacctgct
     4321 ggaggagcgg gacatcaaca gattctccgt gcagaaggac ttcatcaacc ggaatctggt
     4381 ggacaccaga tacgccacac gcggcctgat gaatctgctg cggtcttatt tcagagtgaa
     4441 caatctggat gtgaaggtga agagcatcaa cggcggcttc acctcctttc tgcggagaaa
     4501 gtggaagttt aagaaggagc gcaacaaggg ctataagcac cacgccgagg atgccctgat
     4561 catcgccaat gccgacttca tctttaagga gtggaagaag ctggacaagg ccaagaaagt
     4621 gatggagaac cagatgttcg aggagaagca ggccgagagc atgcccgaga tcgagacaga
     4681 gcaggagtac aaggagattt tcatcacacc tcaccagatc aagcacatca aggacttcaa
     4741 ggactacaag tattctcaca gggtggataa gaagcccaac cgcaagctga tcaatgacac
     4801 cctgtatagc acacggaagg acgataaggg caataccctg atcgtgaaca atctgaacgg
     4861 cctgtacgac aaggataatg acaagctgaa gaagctgatc aacaagtctc ccgagaagct
     4921 gctgatgtac caccacgatc ctcagacata tcagaagctg aagctgatca tggagcagta
     4981 cggcgacgag aagaacccac tgtataagta ctatgaggag acaggcaact acctgacaaa
     5041 gtatagcaag aaggataatg gccccgtgat caagaagatc aagtactatg gcaacaagct
     5101 gaatgcccac ctggacatca ccgacgatta ccctaactct cgcaataagg tggtgaagct
     5161 gagcctgaag ccataccggt tcgacgtgta cctggacaac ggcgtgtata agtttgtgac
     5221 agtgaagaat ctggatgtga tcaagaagga gaactactat gaggtgaaca gcaagtgcta
     5281 cgaggaggcc aagaagctga agaagatcag caaccaggcc gagttcatcg cctcttttta
     5341 caagaatgac ctgatcaaga tcaatggcga gctgtataga gtgatcggcg tgaacaatga
     5401 tctgctgaac agaatcgaag tgaatatgat cgacatcacc tacagggagt atctggagaa
     5461 catgaatgat aagaggcccc ctcatatcat caagaccatc gcctctaaga cacagagcat
     5521 caagaagtac agcacagaca tcctggggaa cctgtatgaa gtcaagagca agaaacatcc
     5581 tcagattatc aagaaaggca gcggcgggag cggcgggagc ggggggagca ctaatctgag
     5641 cgacatcatt gagaaggaga ctgggaaaca gctggtcatt caggagtcca tcctgatgct
     5701 gcctgaggag gtggaggaag tgatcggcaa caagccagag tctgacatcc tggtgcacac
     5761 cgcctacgac gagtccacag atgagaatgt gatgctgctg acctctgacg cccccgagta
     5821 taagccttgg gccctggtca tccaggattc taacggcgag aataagatca agatgctgag
     5881 cggaggatcc aaaagaaccg ccgacggcag cgaattcgag cccaagaaga agaggaaagt
     5941 ctaagatctg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     6001 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     6061 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt agttcttgcc
     6121 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     6181 actgacaatt ccgtggtgcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct
     6241 cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg
     6301 aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc
     6361 aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct
     6421 ctatggctcg agaaaaaaat ctcgccaaca agttgacgag ataaacacgg cattttgcct
     6481 tgttttagta gattctgtaa ttttcattac agagtactaa aaccgagacg cattaatcgt
     6541 ctcgggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag
     6601 ttacggtaag catatgatag tccattttaa aacataattt taaaactgca aactacccaa
     6661 gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat
     6721 taattctaat tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg
     6781 gaaataggcc ctcttcctgc ccgaccttgc ggccgcagga acccctagtg atggagttgg
//
