LOCUS       sequence-368933-        6669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gp41-1 Split N-terminal half of ABEmaxNGA driven by MHP1 promoter 
            with gRNA targeting mdx4cv nonsense mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6669)
  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 6669)
  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: pX601‐MHP1‐miniABEmaxNG‐N2‐(GG)
FEATURES             Location/Qualifiers
     source          1..6669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             706..726
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1327..3042
                     /codon_start=1
                     /product="N-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(N)
                     /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
                     LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
                     LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
                     RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
                     NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
                     GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
                     QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
                     KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
                     SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
                     TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE"
     primer_bind     complement(3340..3357)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3346..3553
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3561..3809
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3561..3581
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3732..3751
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   3932..4072
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3932..4061
                     /label=AAV2 ITR
     rep_origin      4147..4602
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4234..4253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4444..4465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4619..4638)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4738..4760
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4798..4816)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4884..4988
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4989..5057,5058..5849)
                     /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(5207..5226)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6020..6608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6509..6528
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcctcta gattagggtt agggttaggg ttagggctcg aggcccttca
      181 gattaaaaat aactgaggta agggcctggg taggggaggt ggtgtgagac gctcctgtct
      241 ctcctctatc tgcccatcgg ccctttgggg aggaggaatg tgcccaagga ctaaaaaaag
      301 gccatggagc cagaggggcg agggcaacag acctttcatg ggcaaacctt ggggccctgc
      361 tgactgtaga tgagagcagc cactacgggt ctaggctgcc catgtaagga ggcaaggcct
      421 ggggacaccc gagatgcctg gttataatta acccagacat gtggctgccc ccccccccca
      481 acacctgctg cctgctaaaa ataaccctgt ccctggtggc cctgcatgcc ctccctgggg
      541 acagcccctc ctggctagtc acaccctgta ggctcctcta tataacccag gggcacaggg
      601 gctgccctca ttctaccacc acctccacag cacagacaga cactcaggag ccagcccacc
      661 atggctagca tgaaacggac agccgacgga agcgagttcg agtcaccaaa gaagaagcgg
      721 aaagtcggat cctctgaggt ggagttttcc cacgagtact ggatgagaca tgccctgacc
      781 ctggccaaga gggcacgcga tgagagggag gtgcctgtgg gagccgtgct ggtgctgaac
      841 aatagagtga tcggcgaggg ctggaacaga gccatcggcc tgcacgaccc aacaggccat
      901 gccgaaatta tggccctgag gcagggcggc ctggtcatgc agaactacag actgattgac
      961 gccaccctgt acgggacatt cgagccttgc gtgatgtgcg ccggcgccat gatccactct
     1021 aggatcggcc gcgtggtgtt tggcgtgagg aacgcaaaaa ccggcgccgc aggctccctg
     1081 atggacgtgc tgcactaccc cggcatgaat caccgcgtcg aaattaccga gggaatcctg
     1141 gcagatgaat gtgccgccct gctgtgctat ttctttcgga tgcctagaca ggtgttcaat
     1201 gctcagaaga aggcccagag ctccaccgac tccggaggat ctagcggagg ctcctctggc
     1261 tctgagacac ctggcacaag cgagagcgca acacctgaaa gcagcggggg cagcagcggg
     1321 gggtcagaca agaagtacag catcggcctg gccatcggca ccaactctgt gggctgggcc
     1381 gtgatcaccg acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac
     1441 cggcacagca tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc
     1501 gaggccaccc ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc
     1561 tgctatctgc aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac
     1621 agactggaag agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc
     1681 ggcaacatcg tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga
     1741 aagaaactgg tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc
     1801 cacatgatca agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc
     1861 gacgtggaca agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac
     1921 cccatcaacg ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc
     1981 agacggctgg aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga
     2041 aacctgattg ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc
     2101 gaggatgcca aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg
     2161 gcccagatcg gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc
     2221 atcctgctga gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc
     2281 tctatgatca agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg
     2341 cggcagcagc tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac
     2401 gccggctaca ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc
     2461 ctggaaaaga tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg
     2521 cggaagcagc ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg
     2581 cacgccattc tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag
     2641 atcgagaaga tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac
     2701 agcagattcg cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag
     2761 gaagtggtgg acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat
     2821 aagaacctgc ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc
     2881 gtgtataacg agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc
     2941 ctgagcggcg agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg
     3001 accgtgaagc agctgaaaga ggactacttc aagaaaatcg agtgtttgga tctgaaaacg
     3061 caagttcaaa cgccacaggg tatgaaagaa atatccaata tacaggtcgg cgatctcgtc
     3121 ttgtctaaca ctggctataa cgaggtgctg aatgtatttc caaaaagcaa gaaaaaaagt
     3181 tacaagataa ctctggaaga tggaaaagaa attatctgtt ctgaggagca tctgtttccg
     3241 acccaaacag gggagatgaa tatcagtggc ggtctcaaag agggtatgtg tttgtatgtc
     3301 aaggaataac tagtagaatt cctagagctc gctgatcagc ctcgactgtg ccttctagtt
     3361 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
     3421 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
     3481 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gagaatagca
     3541 ggcatgctgg ggaggtacct gagggcctat ttcccatgat tccttcatat ttgcatatac
     3601 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
     3661 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
     3721 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
     3781 tatatatctt gtggaaagga cgaaacaccg gttatctcct gttctgcagc gtttcagagc
     3841 tatgctggaa acagcatagc aagttgaaat aaggctagtc cgttatcaac ttgaaaaagt
     3901 ggcaccgagt cggtgctttt tttgcggccg caggaacccc tagtgatgga gttggccact
     3961 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4021 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     4081 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4141 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4201 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     4261 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4321 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4381 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4441 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     4501 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     4561 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4621 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4681 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4741 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4801 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4861 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4921 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4981 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5041 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5101 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5161 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5221 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5281 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5341 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5401 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5461 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5521 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5581 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5641 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5701 gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5761 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5821 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5881 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5941 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6001 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6061 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6121 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6181 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6241 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6301 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6361 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6421 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6481 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6541 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6601 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6661 ctcacatgt
//
