LOCUS       sequence-368932-        6575 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gp41-1 Split C-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 6575)
  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 6575)
  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‐ABEmaxC2‐NG‐E53 ogRNA
FEATURES             Location/Qualifiers
     source          1..6575
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   30..159
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             794..3178
                     /codon_start=1
                     /product="C-terminal portion of Streptococcus pyogenes Cas9
                     (Zetsche et al., 2015)"
                     /label=Cas9(C)
                     /translation="CFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILED
                     IVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK
                     TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKG
                     ILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQI
                     LKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDN
                     KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDK
                     AGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF
                     YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGK
                     ATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMP
                     QVNIVKKTEVQTGGFSKESIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKV
                     EKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELEN
                     GRKRMLASARFLQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLD
                     EIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKY
                     FDTTIDRKVYRSTKEVLDATLIHQSITGLYETRIDLSQLGGD"
     CDS             3221..3241
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3275..3292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3281..3488
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3496..3744
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3496..3516
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3667..3686
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   3867..4007
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3867..3996
                     /label=AAV2 ITR
     rep_origin      4082..4537
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4169..4188)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4379..4400
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4554..4573)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4673..4695
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4733..4751)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4819..4923
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4924..4992,4993..5784)
                     /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(5142..5161)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5955..6543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6444..6463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac
       61 tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
      121 gcagagaggg agtggccaac tccatcacta ggggttcctg cggcctctag actcgaggcc
      181 cttcagatta aaaataactg aggtaagggc ctgggtaggg gaggtggtgt gagacgctcc
      241 tgtctctcct ctatctgccc atcggccctt tggggaggag gaatgtgccc aaggactaaa
      301 aaaaggccat ggagccagag gggcgagggc aacagacctt tcatgggcaa accttggggc
      361 cctgctgact gtagatgaga gcagccacta cgggtctagg ctgcccatgt aaggaggcaa
      421 ggcctgggga cacccgagat gcctggttat aattaaccca gacatgtggc tgcccccccc
      481 cccaacacct gctgcctgct aaaaataacc ctgtccctgg tggccctgca tgccctccct
      541 ggggacagcc cctcctggct agtcacaccc tgtaggctcc tctatataac ccaggggcac
      601 aggggctgcc ctcattctac caccacctcc acagcacaga cagacactca ggagccagct
      661 agccaccatg atgctcaaga agatcctcaa gattgaagag ttggacgagc gcgagcttat
      721 agacatagaa gtcagtggta atcacctttt ctacgcaaat gacattttga ctcacaactc
      781 ctcttcagac gtttgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc
      841 ctccctgggc acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa
      901 tgaggaaaac gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag
      961 agagatgatc gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa
     1021 gcagctgaag cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg
     1081 catccgggac aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc
     1141 caacagaaac ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca
     1201 gaaagcccag gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg
     1261 cagccccgcc attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa
     1321 agtgatgggc cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac
     1381 cacccagaag ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa
     1441 agagctgggc agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga
     1501 gaagctgtac ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga
     1561 catcaaccgg ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga
     1621 cgactccatc gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa
     1681 cgtgccctcc gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc
     1741 caagctgatt acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag
     1801 cgaactggat aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa
     1861 gcacgtggca cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct
     1921 gatccgggaa gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga
     1981 tttccagttt tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct
     2041 gaacgccgtc gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt
     2101 gtacggcgac tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat
     2161 cggcaaggct accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga
     2221 gattaccctg gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac
     2281 cggggagatc gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat
     2341 gccccaagtg aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc
     2401 tatccggccc aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa
     2461 gaagtacggc ggcttcgtca gccccaccgt ggcctattct gtgctggtgg tggccaaagt
     2521 ggaaaagggc aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat
     2581 ggaaagaagc agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga
     2641 agtgaaaaag gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg
     2701 ccggaagaga atgctggcct ctgcccgctt tctgcagaag ggaaacgaac tggccctgcc
     2761 ctccaaatat gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc
     2821 cgaggataat gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat
     2881 catcgagcag atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa
     2941 agtgctgtcc gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat
     3001 catccacctg tttaccctga ccaatctggg agcccctcgg gccttcaagt actttgacac
     3061 caccatcgac cggaaggtgt accggagcac caaagaggtg ctggacgcca ccctgatcca
     3121 ccagagcatc accggcctgt acgagacacg gatcgacctg tctcagctgg gaggtgactc
     3181 tggcggctca aaaagaaccg ccgacggcag cgaattcgag cccaagaaga agaggaaagt
     3241 ctaactagta gaattcctag agctcgctga tcagcctcga ctgtgccttc tagttgccag
     3301 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     3361 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     3421 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagagaa tagcaggcat
     3481 gctggggagg tacctgaggg cctatttccc atgattcctt catatttgca tatacgatac
     3541 aaggctgtta gagagataat tggaattaat ttgactgtaa acacaaagat attagtacaa
     3601 aatacgtgac gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt
     3661 aaaatggact atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat
     3721 atcttgtgga aaggacgaaa caccggttat ctcctgttct gcagcgtttc agagctatgc
     3781 tggaaacagc atagcaagtt gaaataaggc tagtccgtta tcaacttgaa aaagtggcac
     3841 cgagtcggtg ctttttttgc ggccgcagga acccctagtg atggagttgg ccactccctc
     3901 tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt
     3961 tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc gcctgatgcg
     4021 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag
     4081 tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     4141 gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     4201 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4261 agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg
     4321 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     4381 ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc ttttgattta
     4441 taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta acaaaaattt
     4501 aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct cagtacaatc
     4561 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     4621 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     4681 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg
     4741 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4801 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     4861 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     4921 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     4981 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5041 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5101 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5161 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5221 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5281 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5341 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5401 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5461 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5521 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5581 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtgga
     5641 agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5701 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5761 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     5821 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     5881 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     5941 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6001 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6061 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6121 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6181 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6241 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6301 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6361 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6421 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6481 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6541 aaaaacgcca gcaacgcggc ctttttacgg ttcct
//
