LOCUS       sequence-148034-        4712 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pZac2.1 CMV173-SaCas9-U6-SaDMDR7-U6-SaDMDL2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4712)
  AUTHORS   Tabebordbar M, Zhu K, Cheng JK, Chew WL, Widrick JJ, Yan WX, Maesner
            C, Wu EY, Xiao R, Ran FA, Cong L, Zhang F, Vandenberghe LH, Church 
            GM, Wagers AJ
  TITLE     In vivo gene editing in dystrophic mouse muscle and muscle stem 
            cells.
  JOURNAL   Science. 2016 Jan 22;351(6271):407-11. doi: 10.1126/science.aad5177.
            Epub 2015 Dec 31.
  PUBMED    26721686
REFERENCE   2  (bases 1 to 4712)
  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: 
            "10.1126/science.aad5177"; journalName: "Science"; date: 
            "2016-01-22- 22"; volume: "351"; issue: "6271"; pages: "407-11"
COMMENT     Sequence Label: pZac2.1 CMV173-SaCas9-U6-SaDMDR7-U6-SaDMDL2
FEATURES             Location/Qualifiers
     source          1..4712
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   16..145
                     /label=AAV2 ITR
     promoter        260..359
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     310..330
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             388..3546
                     /codon_start=1
                     /product="Cas9 endonuclease from the Staphylococcus aureus 
                     Type II CRISPR/Cas system"
                     /label=SaCas9
                     /note="generates RNA-guided double strand breaks in DNA"
                     /translation="MKRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNE
                     GRRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEE
                     FSAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGE
                     VRGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFG
                     WKDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKF
                     QIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKE
                     IIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKA
                     INLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSI
                     KVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENA
                     KYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK
                     QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRF
                     SVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKER
                     NKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEI
                     FITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKD
                     NDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKK
                     DNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN
                     LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLL
                     NRIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQ
                     IIKKG"
     CDS             3550..3570
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3574..3594
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3598..3618
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3625..3651
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3661..3782
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3698..3717)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3752..3771
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3789..4037
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     promoter        4158..4406
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     repeat_region   4577..4706
                     /label=AAV2 ITR
ORIGIN
        1 aaggccttaa ttaggctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
       61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      121 gccaactcca tcactagggg ttccttgtag ttaatgatta acccgccatg ctacttatct
      181 acgaattcga gctcggtacc cactcacggg gatttccaag tctccacccc attgacgtca
      241 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      301 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
      361 tctggctaac tccatgccgg tgccaccatg aaaaggaact acattctggg gctggacatc
      421 gggattacaa gcgtggggta tgggattatt gactatgaaa caagggacgt gatcgacgca
      481 ggcgtcagac tgttcaagga ggccaacgtg gaaaacaatg agggacggag aagcaagagg
      541 ggagccaggc gcctgaaacg acggagaagg cacagaatcc agagggtgaa gaaactgctg
      601 ttcgattaca acctgctgac cgaccattct gagctgagtg gaattaatcc ttatgaagcc
      661 agggtgaaag gcctgagtca gaagctgtca gaggaagagt tttccgcagc tctgctgcac
      721 ctggctaagc gccgaggagt gcataacgtc aatgaggtgg aagaggacac cggcaacgag
      781 ctgtctacaa aggaacagat ctcacgcaat agcaaagctc tggaagagaa gtatgtcgca
      841 gagctgcagc tggaacggct gaagaaagat ggcgaggtga gagggtcaat taataggttc
      901 aagacaagcg actacgtcaa agaagccaag cagctgctga aagtgcagaa ggcttaccac
      961 cagctggatc agagcttcat cgatacttat atcgacctgc tggagactcg gagaacctac
     1021 tatgagggac caggagaagg gagccccttc ggatggaaag acatcaagga atggtacgag
     1081 atgctgatgg gacattgcac ctattttcca gaagagctga gaagcgtcaa gtacgcttat
     1141 aacgcagatc tgtacaacgc cctgaatgac ctgaacaacc tggtcatcac cagggatgaa
     1201 aacgagaaac tggaatacta tgagaagttc cagatcatcg aaaacgtgtt taagcagaag
     1261 aaaaagccta cactgaaaca gattgctaag gagatcctgg tcaacgaaga ggacatcaag
     1321 ggctaccggg tgacaagcac tggaaaacca gagttcacca atctgaaagt gtatcacgat
     1381 attaaggaca tcacagcacg gaaagaaatc attgagaacg ccgaactgct ggatcagatt
     1441 gctaagatcc tgactatcta ccagagctcc gaggacatcc aggaagagct gactaacctg
     1501 aacagcgagc tgacccagga agagatcgaa cagattagta atctgaaggg gtacaccgga
     1561 acacacaacc tgtccctgaa agctatcaat ctgattctgg atgagctgtg gcatacaaac
     1621 gacaatcaga ttgcaatctt taaccggctg aagctggtcc caaaaaaggt ggacctgagt
     1681 cagcagaaag agatcccaac cacactggtg gacgatttca ttctgtcacc cgtggtcaag
     1741 cggagcttca tccagagcat caaagtgatc aacgccatca tcaagaagta cggcctgccc
     1801 aatgatatca ttatcgagct ggctagggag aagaacagca aggacgcaca gaagatgatc
     1861 aatgagatgc agaaacgaaa ccggcagacc aatgaacgca ttgaagagat tatccgaact
     1921 accgggaaag agaacgcaaa gtacctgatt gaaaaaatca agctgcacga tatgcaggag
     1981 ggaaagtgtc tgtattctct ggaggccatc cccctggagg acctgctgaa caatccattc
     2041 aactacgagg tcgatcatat tatccccaga agcgtgtcct tcgacaattc ctttaacaac
     2101 aaggtgctgg tcaagcagga agagaactct aaaaagggca ataggactcc tttccagtac
     2161 ctgtctagtt cagattccaa gatctcttac gaaaccttta aaaagcacat tctgaatctg
     2221 gccaaaggaa agggccgcat cagcaagacc aaaaaggagt acctgctgga agagcgggac
     2281 atcaacagat tctccgtcca gaaggatttt attaaccgga atctggtgga cacaagatac
     2341 gctactcgcg gcctgatgaa tctgctgcga tcctatttcc gggtgaacaa tctggatgtg
     2401 aaagtcaagt ccatcaacgg cgggttcaca tcttttctga ggcgcaaatg gaagtttaaa
     2461 aaggagcgca acaaagggta caagcaccat gccgaagatg ctctgattat cgcaaatgcc
     2521 gacttcatct ttaaggagtg gaaaaagctg gacaaagcca agaaagtgat ggagaaccag
     2581 atgttcgaag agaagcaggc cgaatctatg cccgaaatcg agacagaaca ggagtacaag
     2641 gagattttca tcactcctca ccagatcaag catatcaagg atttcaagga ctacaagtac
     2701 tctcaccggg tggataaaaa gcccaacaga gagctgatca atgacaccct gtatagtaca
     2761 agaaaagacg ataaggggaa taccctgatt gtgaacaatc tgaacggact gtacgacaaa
     2821 gataatgaca agctgaaaaa gctgatcaac aaaagtcccg agaagctgct gatgtaccac
     2881 catgatcctc agacatatca gaaactgaag ctgattatgg agcagtacgg cgacgagaag
     2941 aacccactgt ataagtacta tgaagagact gggaactacc tgaccaagta tagcaaaaag
     3001 gataatggcc ccgtgatcaa gaagatcaag tactatggga acaagctgaa tgcccatctg
     3061 gacatcacag acgattaccc taacagtcgc aacaaggtgg tcaagctgtc actgaagcca
     3121 tacagattcg atgtctatct ggacaacggc gtgtataaat ttgtgactgt caagaatctg
     3181 gatgtcatca aaaaggagaa ctactatgaa gtgaatagca agtgctacga agaggctaaa
     3241 aagctgaaaa agattagcaa ccaggcagag ttcatcgcct ccttttacaa caacgacctg
     3301 attaagatca atggcgaact gtatagggtc atcggggtga acaatgatct gctgaaccgc
     3361 attgaagtga atatgattga catcacttac cgagagtatc tggaaaacat gaatgataag
     3421 cgcccccctc gaattatcaa aacaattgcc tctaagactc agagtatcaa aaagtactca
     3481 accgacattc tgggaaacct gtatgaggtg aagagcaaaa agcaccctca gattatcaaa
     3541 aagggcgacc ccaagaagaa gaggaaggtt gatccaaaaa agaagagaaa ggtagatccc
     3601 aagaaaaaaa gaaaagtcgg atcctaccca tacgatgttc cagattacgc ttaagaattc
     3661 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     3721 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     3781 taggtaccga gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg
     3841 ttagagagat aattagaatt aatttgactg taaacacaaa gatattagta caaaatacgt
     3901 gacgtagaaa gtaataattt cttgggtagt ttgcagtttt aaaattatgt tttaaaatgg
     3961 actatcatat gcttaccgta acttgaaagt atttcgattt cttggcttta tatatcttgt
     4021 ggaaaggacg aaacaccgca gtaatgtgtc ataccttcgt ttaagtactc tgtgctggaa
     4081 acagcacaga atctacttaa acaaggcaaa atgccgtgtt tatctcgtca acttgttggc
     4141 gagatttttt tccgcgggag ggcctatttc ccatgattcc ttcatatttg catatacgat
     4201 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
     4261 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
     4321 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
     4381 atatcttgtg gaaaggacga aacaccgata taatagaaat tattcatgtt taagtactct
     4441 gtgctggaaa cagcacagaa tctacttaaa caaggcaaaa tgccgtgttt atctcgtcaa
     4501 cttgttggcg agattttttt ggggatcctc tagagtcgag tagataagta gcatggcggg
     4561 ttaatcatta actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct
     4621 cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg
     4681 gcctcagtga gcgagcgagc gcgcagcctt aa
//
