LOCUS       sequence-368936-        6702 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gp41-1 Split N-terminal half of ABEmaxNGA with gRNA targeting mdx4cv
            nonsense mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6702)
  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 6702)
  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‐mhCMV‐miniABEmax‐N2‐ E53ogRNA
FEATURES             Location/Qualifiers
     source          1..6702
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   17..146
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        582..681
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     632..652
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      704..713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             755..775
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1376..3091
                     /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(3389..3406)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3395..3602
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3610..3858
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     3610..3630
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3781..3800
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     repeat_region   3981..4121
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3981..4110
                     /label=AAV2 ITR
     rep_origin      4196..4651
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4283..4302)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4493..4514
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4668..4687)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4787..4809
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4847..4865)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4933..5037
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5038..5106,5107..5898)
                     /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(5256..5275)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6069..6657
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6558..6577
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
       61 gcgtcgggcg acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt
      121 ggccaactcc atcactaggg gttcctgcgg cctctagact cgaggccctt cagattaaaa
      181 ataactgagg taagggcctg ggtaggggag gtggtgtgag acgctcctgt ctctcctcta
      241 tctgcccatc ggccctttgg ggaggaggaa tgtgcccaag gactaaaaaa aggccatgga
      301 gccagagggg cgagggcaac agacctttca tgggcaaacc ttggggccct gctgactgta
      361 gatgagagca gccactacgg gtctaggctg cccatgtaag gaggcaaggc ctggggacac
      421 ccgagatgcc tggttataat taacccagac atgtggctgc cccccccccc naacacctgc
      481 tgcctgctaa aaataaccct gtccctggtg gccctgcatg cccactcacg gggatttcca
      541 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      601 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      661 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgccacca tggctagcat
      721 gaaacggaca gccgacggaa gcgagttcga gtcaccaaag aagaagcgga aagtcggatc
      781 ctctgaggtg gagttttccc acgagtactg gatgagacat gccctgaccc tggccaagag
      841 ggcacgcgat gagagggagg tgcctgtggg agccgtgctg gtgctgaaca atagagtgat
      901 cggcgagggc tggaacagag ccatcggcct gcacgaccca acaggccatg ccgaaattat
      961 ggccctgagg cagggcggcc tggtcatgca gaactacaga ctgattgacg ccaccctgta
     1021 cgggacattc gagccttgcg tgatgtgcgc cggcgccatg atccactcta ggatcggccg
     1081 cgtggtgttt ggcgtgagga acgcaaaaac cggcgccgca ggctccctga tggacgtgct
     1141 gcactacccc ggcatgaatc accgcgtcga aattaccgag ggaatcctgg cagatgaatg
     1201 tgccgccctg ctgtgctatt tctttcggat gcctagacag gtgttcaatg ctcagaagaa
     1261 ggcccagagc tccaccgact ccggaggatc tagcggaggc tcctctggct ctgagacacc
     1321 tggcacaagc gagagcgcaa cacctgaaag cagcgggggc agcagcgggg ggtcagacaa
     1381 gaagtacagc atcggcctgg ccatcggcac caactctgtg ggctgggccg tgatcaccga
     1441 cgagtacaag gtgcccagca agaaattcaa ggtgctgggc aacaccgacc ggcacagcat
     1501 caagaagaac ctgatcggag ccctgctgtt cgacagcggc gaaacagccg aggccacccg
     1561 gctgaagaga accgccagaa gaagatacac cagacggaag aaccggatct gctatctgca
     1621 agagatcttc agcaacgaga tggccaaggt ggacgacagc ttcttccaca gactggaaga
     1681 gtccttcctg gtggaagagg ataagaagca cgagcggcac cccatcttcg gcaacatcgt
     1741 ggacgaggtg gcctaccacg agaagtaccc caccatctac cacctgagaa agaaactggt
     1801 ggacagcacc gacaaggccg acctgcggct gatctatctg gccctggccc acatgatcaa
     1861 gttccggggc cacttcctga tcgagggcga cctgaacccc gacaacagcg acgtggacaa
     1921 gctgttcatc cagctggtgc agacctacaa ccagctgttc gaggaaaacc ccatcaacgc
     1981 cagcggcgtg gacgccaagg ccatcctgtc tgccagactg agcaagagca gacggctgga
     2041 aaatctgatc gcccagctgc ccggcgagaa gaagaatggc ctgttcggaa acctgattgc
     2101 cctgagcctg ggcctgaccc ccaacttcaa gagcaacttc gacctggccg aggatgccaa
     2161 actgcagctg agcaaggaca cctacgacga cgacctggac aacctgctgg cccagatcgg
     2221 cgaccagtac gccgacctgt ttctggccgc caagaacctg tccgacgcca tcctgctgag
     2281 cgacatcctg agagtgaaca ccgagatcac caaggccccc ctgagcgcct ctatgatcaa
     2341 gagatacgac gagcaccacc aggacctgac cctgctgaaa gctctcgtgc ggcagcagct
     2401 gcctgagaag tacaaagaga ttttcttcga ccagagcaag aacggctacg ccggctacat
     2461 tgacggcgga gccagccagg aagagttcta caagttcatc aagcccatcc tggaaaagat
     2521 ggacggcacc gaggaactgc tcgtgaagct gaacagagag gacctgctgc ggaagcagcg
     2581 gaccttcgac aacggcagca tcccccacca gatccacctg ggagagctgc acgccattct
     2641 gcggcggcag gaagattttt acccattcct gaaggacaac cgggaaaaga tcgagaagat
     2701 cctgaccttc cgcatcccct actacgtggg ccctctggcc aggggaaaca gcagattcgc
     2761 ctggatgacc agaaagagcg aggaaaccat caccccctgg aacttcgagg aagtggtgga
     2821 caagggcgct tccgcccaga gcttcatcga gcggatgacc aacttcgata agaacctgcc
     2881 caacgagaag gtgctgccca agcacagcct gctgtacgag tacttcaccg tgtataacga
     2941 gctgaccaaa gtgaaatacg tgaccgaggg aatgagaaag cccgccttcc tgagcggcga
     3001 gcagaaaaag gccatcgtgg acctgctgtt caagaccaac cggaaagtga ccgtgaagca
     3061 gctgaaagag gactacttca agaaaatcga gtgtttggat ctgaaaacgc aagttcaaac
     3121 gccacagggt atgaaagaaa tatccaatat acaggtcggc gatctcgtct tgtctaacac
     3181 tggctataac gaggtgctga atgtatttcc aaaaagcaag aaaaaaagtt acaagataac
     3241 tctggaagat ggaaaagaaa ttatctgttc tgaggagcat ctgtttccga cccaaacagg
     3301 ggagatgaat atcagtggcg gtctcaaaga gggtatgtgt ttgtatgtca aggaataact
     3361 agtagaattc ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct
     3421 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3481 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3541 ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag gcatgctggg
     3601 gaggtacctg agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct
     3661 gttagagaga taattggaat taatttgact gtaaacacaa agatattagt acaaaatacg
     3721 tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg ttttaaaatg
     3781 gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt atatatcttg
     3841 tggaaaggac gaaacaccgg ttatctcctg ttctgcagcg tttcagagct atgctggaaa
     3901 cagcatagca agttgaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
     3961 ggtgcttttt ttgcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     4021 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4081 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     4141 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4201 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4261 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4321 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4381 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4441 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4501 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     4561 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4621 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4681 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4741 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4801 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4861 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4921 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4981 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5041 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5101 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5161 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5221 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5281 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5341 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5401 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5461 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5521 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5581 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5641 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5701 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc
     5761 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5821 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5881 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5941 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6001 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6061 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6121 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6181 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6241 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6301 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6361 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6421 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6481 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6541 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6601 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6661 gccagcaacg cggccttttt acggttcctg gccttttgct gg
//
