LOCUS       sequence-368550-        8217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ABE8e(V106W)-SpG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8217)
  AUTHORS   Xu L, Zhang C, Li H, Wang P, Gao Y, Mokadam NA, Ma J, Arnold WD, Han
            R
  TITLE     Efficient precise in vivo base editing in adult dystrophic mice.
  JOURNAL   Nat Commun. 2021 Jun 17;12(1):3719. doi: 10.1038/s41467-021-23996-y.
  PUBMED    34140489
REFERENCE   2  (bases 1 to 8217)
  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 
            Commun."; date: "2021-06-17"; volume: "12(1):3719. doi"; pages: " 
            10.1038/s41467-021-23996-y"
COMMENT     Sequence Label: pCMV_SpG‐ABE8e(V106W)
FEATURES             Location/Qualifiers
     source          1..8217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(28..47)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        219..598
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        599..802
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     753..773
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     844..863
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        844..862
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             912..932
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1527..5627
                     /codon_start=1
                     /product="Cas9 (Csn1) endonuclease from the Streptococcus 
                     pyogenes Type II CRISPR/Cas system"
                     /label=Cas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
                     VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
                     KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
                     LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
                     KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
                     YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
                     EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
                     AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
                     AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
                     ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
                     SILPKRNSDKLIARKKDWDPKKYGGFLWPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
                     IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAKQLQKGNEL
                     ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
                     NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVLD
                     ATLIHQSITGLYETRIDLSQLGGD"
     CDS             5670..5690
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             5699..5716
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5739..5756)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5745..5969
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6040..6056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6040..6056)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6053..6075)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6064..6080
                     /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)."
     promoter        complement(join(6088..6094,6095..6112,6113..6118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6133..6154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6271..6288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6442..7030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6522..6541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7201..7992,7993..8061))
                     /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     7824..7843
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8062..8166)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcccgatccc ctagggtcga ctctcagtac aatctgctct gatgccgcat agttaagcca
       61 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
      121 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
      181 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      241 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatattgagt tccgcgttac
      301 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      361 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      421 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      481 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      541 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      601 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      661 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      721 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      781 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctaga gatccgcggc
      841 cgctaatacg actcactata gggagagccg ccaccatgaa acggacagcc gacggaagcg
      901 agttcgagtc accaaagaag aagcggaaag tctctgaggt ggagttttcc cacgagtact
      961 ggatgagaca tgccctgacc ctggccaaga gggcacggga tgagagggag gtgcctgtgg
     1021 gagccgtgct ggtgctgaac aatagagtga tcggcgaggg ctggaacaga gccatcggcc
     1081 tgcacgaccc aacagcccat gccgaaatta tggccctgag acagggcggc ctggtcatgc
     1141 agaactacag actgattgac gccaccctgt acgtgacatt cgagccttgc gtgatgtgcg
     1201 ccggcgccat gatccactct aggatcggcc gcgtggtgtt tggatggaga aattctaaaa
     1261 gaggcgccgc aggctccctg atgaacgtgc tgaactaccc cggcatgaat caccgcgtcg
     1321 aaattaccga gggaatcctg gcagatgaat gtgccgccct gctgtgcgat ttctatcgga
     1381 tgcctagaca ggtgttcaat gctcagaaga aggcccagag ctccatcaac tccggaggat
     1441 ctagcggagg ctcctctggc tctgagacac ctggcacaag cgagagcgca acacctgaaa
     1501 gcagcggggg cagcagcggg gggtcagaca agaagtacag catcggcctg gccatcggca
     1561 ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc aagaaattca
     1621 aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga gccctgctgt
     1681 tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga agaagataca
     1741 ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag atggccaagg
     1801 tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag gataagaagc
     1861 acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac gagaagtacc
     1921 ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc gacctgcggc
     1981 tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg atcgagggcg
     2041 acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg cagacctaca
     2101 accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag gccatcctgt
     2161 ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg cccggcgaga
     2221 agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc cccaacttca
     2281 agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac acctacgacg
     2341 acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg tttctggccg
     2401 ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac accgagatca
     2461 ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac caggacctga
     2521 ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag attttcttcg
     2581 accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag gaagagttct
     2641 acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc
     2701 tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc atcccccacc
     2761 agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt tacccattcc
     2821 tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc tactacgtgg
     2881 gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc gaggaaacca
     2941 tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag agcttcatcg
     3001 agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc
     3061 tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac gtgaccgagg
     3121 gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt
     3181 tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc aagaaaatcg
     3241 agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc tccctgggca
     3301 cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat gaggaaaacg
     3361 aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga gagatgatcg
     3421 aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag cagctgaagc
     3481 ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc atccgggaca
     3541 agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc aacagaaact
     3601 tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag aaagcccagg
     3661 tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc agccccgcca
     3721 ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa gtgatgggcc
     3781 ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc acccagaagg
     3841 gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa gagctgggca
     3901 gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag aagctgtacc
     3961 tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac atcaaccggc
     4021 tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac gactccatcg
     4081 acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac gtgccctccg
     4141 aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc aagctgatta
     4201 cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc gaactggata
     4261 aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag cacgtggcac
     4321 agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg atccgggaag
     4381 tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat ttccagtttt
     4441 acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg aacgccgtcg
     4501 tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg tacggcgact
     4561 acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc ggcaaggcta
     4621 ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag attaccctgg
     4681 ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc ggggagatcg
     4741 tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg ccccaagtga
     4801 atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct atcctgccca
     4861 agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag aagtacggcg
     4921 gcttcctgtg gcccaccgtg gcctattctg tgctggtggt ggccaaagtg gaaaagggca
     4981 agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg gaaagaagca
     5041 gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa gtgaaaaagg
     5101 acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc cggaagagaa
     5161 tgctggcctc tgccaagcag ctgcagaagg gaaacgaact ggccctgccc tccaaatatg
     5221 tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc gaggataatg
     5281 agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc atcgagcaga
     5341 tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa gtgctgtccg
     5401 cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc atccacctgt
     5461 ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc accatcgacc
     5521 ggaagcagta cagaagcacc aaagaggtgc tggacgccac cctgatccac cagagcatca
     5581 ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggtgactct ggcggctcaa
     5641 aaagaaccgc cgacggcagc gaattcgagc ccaagaagaa gaggaaagtc taaccggtca
     5701 tcatcaccat caccattgag tttaaacccg ctgatcagcc tcgactgtgc cttctagttg
     5761 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     5821 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     5881 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     5941 gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gctggggctc
     6001 gataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6061 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6121 ctagggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6181 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6241 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6301 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6361 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6421 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6481 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6541 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6601 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6661 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6721 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6781 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6841 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6901 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6961 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7021 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacactcagt ggaacgaaaa
     7081 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7141 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     7201 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7261 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7321 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7381 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7441 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7501 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7561 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7621 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7681 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7741 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7801 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7861 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7921 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7981 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8041 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8101 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     8161 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatc
//
