LOCUS       sequence-407964-        3339 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mutagenesis of Cnr1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3339)
  AUTHORS   Gunduz-Cinar O, Castillo LI, Xia M, Van Leer E, Brockway ET, Pollack
            GA, Yasmin F, Bukalo O, Limoges A, Oreizi-Esfahani S, Kondev V, 
            Baldi R, Dong A, Harvey-White J, Cinar R, Kunos G, Li Y, Zweifel LS,
            Patel S, Holmes A
  TITLE     A cortico-amygdala neural substrate for endocannabinoid modulation 
            of fear extinction.
  JOURNAL   Neuron. 2023 Jul 19:S0896-6273(23)00482-8. doi: 
            10.1016/j.neuron.2023.06.023.
  PUBMED    37480845
REFERENCE   2  (bases 1 to 3339)
  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: "Neuron. 
            2023 Jul 19:S0896-6273(23)00482-8. doi: 
            10.1016/j.neuron.2023.06.023."
COMMENT     Sequence Label: pAAV-CMV-FLEX-SaCas9-U6-sgCnr1
FEATURES             Location/Qualifiers
     source          1..3339
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..21
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             46..3201
                     /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="KRNYILGLDIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEG
                     RRSKRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEF
                     SAALLHLAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEV
                     RGSINRFKTSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGW
                     KDIKEWYEMLMGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQ
                     IIENVFKQKKKPTLKQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEI
                     IENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAI
                     NLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFILSPVVKRSFIQSIK
                     VINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTTGKENAK
                     YLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVKQ
                     EENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFS
                     VQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERN
                     KGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIF
                     ITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDN
                     DKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKD
                     NGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNL
                     DVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLN
                     RIEVNMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQI
                     IKKG"
     CDS             3202..3249
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             3256..3336
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA"
ORIGIN
        1 ccaaagaaga agcggaaggt cggtatccac ggagtcccag cagccaagcg gaactacatc
       61 ctgggcctgg acatcggcat caccagcgtg ggctacggca tcatcgacta cgagacacgg
      121 gacgtgatcg atgccggcgt gcggctgttc aaagaggcca acgtggaaaa caacgagggc
      181 aggcggagca agagaggcgc cagaaggctg aagcggcgga ggcggcatag aatccagaga
      241 gtgaagaagc tgctgttcga ctacaacctg ctgaccgacc acagcgagct gagcggcatc
      301 aacccctacg aggccagagt gaagggcctg agccagaagc tgagcgagga agagttctct
      361 gccgccctgc tgcacctggc caagagaaga ggcgtgcaca acgtgaacga ggtggaagag
      421 gacaccggca acgagctgtc caccaaagag cagatcagcc ggaacagcaa ggccctggaa
      481 gagaaatacg tggccgaact gcagctggaa cggctgaaga aagacggcga agtgcggggc
      541 agcatcaaca gattcaagac cagcgactac gtgaaagaag ccaaacagct gctgaaggtg
      601 cagaaggcct accaccagct ggaccagagc ttcatcgaca cctacatcga cctgctggaa
      661 acccggcgga cctactatga gggacctggc gagggcagcc ccttcggctg gaaggacatc
      721 aaagaatggt acgagatgct gatgggccac tgcacctact tccccgagga actgcggagc
      781 gtgaagtacg cctacaacgc cgacctgtac aacgccctga acgacctgaa caatctcgtg
      841 atcaccaggg acgagaacga gaagctggaa tattacgaga agttccagat catcgagaac
      901 gtgttcaagc agaagaagaa gcccaccctg aagcagatcg ccaaagaaat cctcgtgaac
      961 gaagaggata ttaagggcta cagagtgacc agcaccggca agcccgagtt caccaacctg
     1021 aaggtgtacc acgacatcaa ggacattacc gcccggaaag agattattga gaacgccgag
     1081 ctgctggatc agattgccaa gatcctgacc atctaccaga gcagcgagga catccaggaa
     1141 gaactgacca atctgaactc cgagctgacc caggaagaga tcgagcagat ctctaatctg
     1201 aagggctata ccggcaccca caacctgagc ctgaaggcca tcaacctgat cctggacgag
     1261 ctgtggcaca ccaacgacaa ccagatcgct atcttcaacc ggctgaagct ggtgcccaag
     1321 aaggtggacc tgtcccagca gaaagagatc cccaccaccc tggtggacga cttcatcctg
     1381 agccccgtcg tgaagagaag cttcatccag agcatcaaag tgatcaacgc catcatcaag
     1441 aagtacggcc tgcccaacga catcattatc gagctggccc gcgagaagaa ctccaaggac
     1501 gcccagaaaa tgatcaacga gatgcagaag cggaaccggc agaccaacga gcggatcgag
     1561 gaaatcatcc ggaccaccgg caaagagaac gccaagtacc tgatcgagaa gatcaagctg
     1621 cacgacatgc aggaaggcaa gtgcctgtac agcctggaag ccatccctct ggaagatctg
     1681 ctgaacaacc ccttcaacta tgaggtggac cacatcatcc ccagaagcgt gtccttcgac
     1741 aacagcttca acaacaaggt gctcgtgaag caggaagaaa acagcaagaa gggcaaccgg
     1801 accccattcc agtacctgag cagcagcgac agcaagatca gctacgaaac cttcaagaag
     1861 cacatcctga atctggccaa gggcaagggc agaatcagca agaccaagaa agagtatctg
     1921 ctggaagaac gggacatcaa caggttctcc gtgcagaaag acttcatcaa ccggaacctg
     1981 gtggatacca gatacgccac cagaggcctg atgaacctgc tgcggagcta cttcagagtg
     2041 aacaacctgg acgtgaaagt gaagtccatc aatggcggct tcaccagctt tctgcggcgg
     2101 aagtggaagt ttaagaaaga gcggaacaag gggtacaagc accacgccga ggacgccctg
     2161 atcattgcca acgccgattt catcttcaaa gagtggaaga aactggacaa ggccaaaaaa
     2221 gtgatggaaa accagatgtt cgaggaaaag caggccgaga gcatgcccga gatcgaaacc
     2281 gagcaggagt acaaagagat cttcatcacc ccccaccaga tcaagcacat taaggacttc
     2341 aaggactaca agtacagcca ccgggtggac aagaagccta atagagagct gattaacgac
     2401 accctgtact ccacccggaa ggacgacaag ggcaacaccc tgatcgtgaa caatctgaac
     2461 ggcctgtacg acaaggacaa tgacaagctg aaaaagctga tcaacaagag ccccgaaaag
     2521 ctgctgatgt accaccacga cccccagacc taccagaaac tgaagctgat tatggaacag
     2581 tacggcgacg agaagaatcc cctgtacaag tactacgagg aaaccgggaa ctacctgacc
     2641 aagtactcca aaaaggacaa cggccccgtg atcaagaaga ttaagtatta cggcaacaaa
     2701 ctgaacgccc atctggacat caccgacgac taccccaaca gcagaaacaa ggtcgtgaag
     2761 ctgtccctga agccctacag attcgacgtg tacctggaca atggcgtgta caagttcgtg
     2821 accgtgaaga atctggatgt gatcaaaaaa gaaaactact acgaagtgaa tagcaagtgc
     2881 tatgaggaag ctaagaagct gaagaagatc agcaaccagg ccgagtttat cgcctccttc
     2941 tacaacaacg atctgatcaa gatcaacggc gagctgtata gagtgatcgg cgtgaacaac
     3001 gacctgctga accggatcga agtgaacatg atcgacatca cctaccgcga gtacctggaa
     3061 aacatgaacg acaagaggcc ccccaggatc attaagacaa tcgcctccaa gacccagagc
     3121 attaagaagt acagcacaga cattctgggc aacctgtatg aagtgaaatc taagaagcac
     3181 cctcagatca tcaaaaaggg caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa
     3241 aagaaaaagg gatcctaccc atacgatgtt ccagattacg cttacccata cgatgttcca
     3301 gattacgctt acccatacga tgttccagat tacgcttaa
//
