LOCUS       sequence-148037-        4738 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV vector containing SaCas9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4738)
  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 4738)
  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 SV40-CMV-SaCas9-3xNLS
FEATURES             Location/Qualifiers
     source          1..4738
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     complement(186..206)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     enhancer        436..815
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        816..1019
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     970..990
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1053..4211
                     /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             4215..4235
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4239..4259
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4263..4283
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4290..4316
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(4344..4361)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4350..4557
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4609..4738
                     /label=AAV2 ITR
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgcg gcctctagac
      181 tcgagggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
      241 tatgcaaagc atgcatctca attagtcagc aaccagtccc ggtctcctcc catgcatgtc
      301 aatattggcc attagccata ttattcattg gttatatagc ataaatcaat attggctatt
      361 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
      421 tatgaccgcc atgttggcat tgattattga ctagttatta atagtaatca attacggggt
      481 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      541 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      601 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      661 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
      721 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
      781 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
      841 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      901 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
      961 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagaggtc
     1021 gtttagtgaa ccgtcagatc accggtgcca ccatgaaaag gaactacatt ctggggctgg
     1081 acatcgggat tacaagcgtg gggtatggga ttattgacta tgaaacaagg gacgtgatcg
     1141 acgcaggcgt cagactgttc aaggaggcca acgtggaaaa caatgaggga cggagaagca
     1201 agaggggagc caggcgcctg aaacgacgga gaaggcacag aatccagagg gtgaagaaac
     1261 tgctgttcga ttacaacctg ctgaccgacc attctgagct gagtggaatt aatccttatg
     1321 aagccagggt gaaaggcctg agtcagaagc tgtcagagga agagttttcc gcagctctgc
     1381 tgcacctggc taagcgccga ggagtgcata acgtcaatga ggtggaagag gacaccggca
     1441 acgagctgtc tacaaaggaa cagatctcac gcaatagcaa agctctggaa gagaagtatg
     1501 tcgcagagct gcagctggaa cggctgaaga aagatggcga ggtgagaggg tcaattaata
     1561 ggttcaagac aagcgactac gtcaaagaag ccaagcagct gctgaaagtg cagaaggctt
     1621 accaccagct ggatcagagc ttcatcgata cttatatcga cctgctggag actcggagaa
     1681 cctactatga gggaccagga gaagggagcc ccttcggatg gaaagacatc aaggaatggt
     1741 acgagatgct gatgggacat tgcacctatt ttccagaaga gctgagaagc gtcaagtatg
     1801 cttataacgc agatctgtac aacgccctga atgacctgaa caacctggtc atcaccaggg
     1861 atgaaaacga gaaactggaa tactatgaga agttccagat catcgaaaac gtgtttaagc
     1921 agaagaaaaa gcctacactg aaacagattg ctaaggagat cctggtcaac gaagaggaca
     1981 tcaagggcta ccgggtgaca agcactggaa aaccagagtt caccaatctg aaagtgtatc
     2041 acgatattaa ggacatcaca gcacggaaag aaatcattga gaacgccgaa ctgctggatc
     2101 agattgctaa gatcctgact atctaccaga gctccgagga catccaggaa gagctgacta
     2161 acctgaacag cgagctgacc caggaagaga tcgaacagat tagtaatctg aaggggtaca
     2221 ccggaacaca caacctgtcc ctgaaagcta tcaatctgat tctggatgag ctgtggcata
     2281 caaacgacaa tcagattgca atctttaacc ggctgaagct ggtcccaaaa aaggtggacc
     2341 tgagtcagca gaaagagatc ccaaccacac tggtggacga tttcattctg tcacccgtgg
     2401 tcaagcggag cttcatccag agcatcaaag tgatcaacgc catcatcaag aagtacggcc
     2461 tgcccaatga tatcattatc gagctggcta gggagaagaa cagcaaggac gcacagaaga
     2521 tgatcaatga gatgcagaaa cgaaaccggc agaccaatga acgcattgaa gagattatcc
     2581 gaactaccgg gaaagagaac gcaaagtacc tgattgaaaa aatcaagctg cacgatatgc
     2641 aggagggaaa gtgtctgtat tctctggagg ccatccccct ggaggacctg ctgaacaatc
     2701 cattcaacta cgaggtcgat catattatcc ccagaagcgt gtccttcgac aattccttta
     2761 acaacaaggt gctggtcaag caggaagaga actctaaaaa gggcaatagg actcctttcc
     2821 agtacctgtc tagttcagat tccaagatct cttacgaaac ctttaaaaag cacattctga
     2881 atctggccaa aggaaagggc cgcatcagca agaccaaaaa ggagtacctg ctggaagagc
     2941 gggacatcaa cagattctcc gtccagaagg attttattaa ccggaatctg gtggacacaa
     3001 gatacgctac tcgcggcctg atgaatctgc tgcgatccta tttccgggtg aacaatctgg
     3061 atgtgaaagt caagtccatc aacggcgggt tcacatcttt tctgaggcgc aaatggaagt
     3121 ttaaaaagga gcgcaacaaa gggtacaagc accatgccga agatgctctg attatcgcaa
     3181 atgccgactt catctttaag gagtggaaaa agctggacaa agccaagaaa gtgatggaga
     3241 accagatgtt cgaagagaag caggccgaat ctatgcccga aatcgagaca gaacaggagt
     3301 acaaggagat tttcatcact cctcaccaga tcaagcatat caaggatttc aaggactaca
     3361 agtactctca ccgggtggat aaaaagccca acagagagct gatcaatgac accctgtata
     3421 gtacaagaaa agacgataag gggaataccc tgattgtgaa caatctgaac ggactgtacg
     3481 acaaagataa tgacaagctg aaaaagctga tcaacaaaag tcccgagaag ctgctgatgt
     3541 accaccatga tcctcagaca tatcagaaac tgaagctgat tatggagcag tacggcgacg
     3601 agaagaaccc actgtataag tactatgaag agactgggaa ctacctgacc aagtatagca
     3661 aaaaggataa tggccccgtg atcaagaaga tcaagtacta tgggaacaag ctgaatgccc
     3721 atctggacat cacagacgat taccctaaca gtcgcaacaa ggtggtcaag ctgtcactga
     3781 agccatacag attcgatgtc tatctggaca acggcgtgta taaatttgtg actgtcaaga
     3841 atctggatgt catcaaaaag gagaactact atgaagtgaa tagcaagtgc tacgaagagg
     3901 ctaaaaagct gaaaaagatt agcaaccagg cagagttcat cgcctccttt tacaacaacg
     3961 acctgattaa gatcaatggc gaactgtata gggtcatcgg ggtgaacaat gatctgctga
     4021 accgcattga agtgaatatg attgacatca cttaccgaga gtatctggaa aacatgaatg
     4081 ataagcgccc ccctcgaatt atcaaaacaa ttgcctctaa gactcagagt atcaaaaagt
     4141 actcaaccga cattctggga aacctgtatg aggtgaagag caaaaagcac cctcagatta
     4201 tcaaaaaggg cgaccccaag aagaagagga aggttgatcc aaaaaagaag agaaaggtag
     4261 atcccaagaa aaaaagaaaa gtcggatcct acccatacga tgttccagat tacgcttaag
     4321 aattcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4381 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4441 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4501 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg
     4561 gccgctagaa tgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag
     4621 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4681 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcag
//
