LOCUS       sequence-180397-        5742 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of mEGFP and paAIP2 for optogenetic control 
            of CaMKII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5742)
  AUTHORS   Murakoshi H, Shin ME, Parra-Bueno P, Szatmari EM, Shibata AC, Yasuda
            R
  TITLE     Kinetics of Endogenous CaMKII Required for Synaptic Plasticity 
            Revealed by Optogenetic Kinase Inhibitor.
  JOURNAL   Neuron. 2017 Apr 5;94(1):37-47.e5. doi: 
            10.1016/j.neuron.2017.02.036. Epub 2017 Mar 16.
  PUBMED    28318784
REFERENCE   2  (bases 1 to 5742)
  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: 
            "10.1016/j.neuron.2017.02.036"; journalName: "Neuron"; date: 
            "2017-04-5- 5"; volume: "94"; issue: "1"; pages: "37-47.e5"
COMMENT     Sequence Label: CaMKIIP-mEGFP-P2A-paAIP2/pAAV
FEATURES             Location/Qualifiers
     source          1..5742
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   54..183
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        227..1515
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1538..1547
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1544..1546,1547..1549,1550..2260)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1589..1610)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1850..1869)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2197..2218
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2270..2326
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     repeat_region   3058..3198
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3058..3187
                     /label=AAV2 ITR
     rep_origin      3273..3728
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3360..3379)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3570..3591
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3745..3764)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3864..3886
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3924..3942)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4010..4114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4115..4183,4184..4975)
                     /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(4333..4352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5146..5734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5635..5654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag
       61 gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc
      121 cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt
      181 cctgcggccg cacgcgtatt accgccatgc attagttatt aattaacatt atggccttag
      241 gtcacttcat ctccatgggg ttcttcttct gattttctag aaaatgagat gggggtgcag
      301 agagcttcct cagtgacctg cccagggtca catcagaaat gtcagagcta gaacttgaac
      361 tcagattact aatcttaaat tccatgcctt gggggcatgc aagtacgata tacagaagga
      421 gtgaactcat tagggcagat gaccaatgag tttaggaaag aagagtccag ggcagggtac
      481 atctacacca cccgcccagc cctgggtgag tccagccacg ttcacctcat tatagttgcc
      541 tctctccagt cctaccttga cgggaagcac aagcagaaac tgggacagga gccccaggag
      601 accaaatctt catggtccct ctgggaggat gggtggggag agctgtggca gaggcctcag
      661 gaggggccct gctgctcagt ggtgacagat aggggtgaga aagcagacag agtcattccg
      721 tcagcattct gggtctgttt ggtacttctt ctcacgctaa ggtggcggtg tgatatgcac
      781 aatggctaaa aagcagggag agctggaaag aaacaaggac agagacagag gccaagtcaa
      841 ccagaccaat tcccagagga agcaaagaaa ccattacaga gactacaagg gggaagggaa
      901 ggagagatga attagcttcc cctgtaaacc ttagaaccca gctgttgcca gggcaacggg
      961 gcaatacctg tctcttcaga ggagatgaag ttgccagggt aactacatcc tgtctttctc
     1021 aaggaccatc ccagaatgtg gcacccacta gccgttacca tagcaactgc ctctttgccc
     1081 cacttaatcc catcccgtct gttaaaaggg ccctatagtt ggaggtgggg gaggtaggaa
     1141 gagcgatgat cacttgtgga ctaagtttgt tcgcatcccc ttctccaacc ccctcagtac
     1201 atcaccctgg gggaacaggg tccacttgct cctgggccca cacagtcctg cagtattgtg
     1261 tatataaggc cagggcaaag aggagcaggt tttaaagtga aaggcaggca ggtgttgggg
     1321 aggcagttac cggggcaacg ggaacagggc gtttcggagg tggttgccat ggggacctgg
     1381 atgctgacga aggctcgcga ggctgtgagc agccacagtg ccctgctcag aagccccaag
     1441 ctcgtcagtc aagccggttc tccgtttgca ctcaggagca cgggcaggcg agtggcccct
     1501 agttctgggg gcagcggggg atccccgggt accggtcgcc accatggtga gcaagggcga
     1561 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1621 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     1681 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     1741 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     1801 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     1861 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1921 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1981 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     2041 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     2101 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     2161 caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2221 cgccgccggg atcactctcg gcatggacga gctgtacaag ggaagcggag ccactaactt
     2281 ctccctgttg aaacaagcag gggatgtcga agagaatccc gggccaggat ccatgctggc
     2341 gaccaccctg gaacgcattg aaaaaaactt tgtgattacc gatccgcgcc tgccggataa
     2401 cccgattatt tttgcgagcg atagctttct gcagctgacc gaatatagcc gcgaagaaat
     2461 tctgggccgc aactgccgct ttctgcaggg cccggaaacc gatcgcgcga ccgtgcgcaa
     2521 aattcgcgat gcgattgata accagaccga agtgaccgtg cagctgatta actataccaa
     2581 aagcggcaaa aaattttgga acctgtttca tctgcagccg atgcgcgatc agaaaggcga
     2641 tgtgcagtat tttattggcg tgcagctgga tggcaccgaa catgtgcgcg atgcggcgga
     2701 acgcgaaggc gtgatgctga ttaaaaaaac cgcggaaaac attaagaaga aactgagaag
     2761 acaggaagcg tttgatgcgc tgtaaaagct tgtggcatcc ctgtgacccc tccccagtgc
     2821 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     2881 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     2941 tatggagcaa ggggcaagtt gggaagaccc aaattgctgg gattacaggc gtgaaccact
     3001 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg
     3061 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3121 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3181 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3241 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3301 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3361 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3421 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3481 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3541 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3601 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3661 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3721 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3781 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3841 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3901 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3961 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4021 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4081 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4201 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4261 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4321 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4381 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4441 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4501 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4561 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4621 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4681 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4741 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4801 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     4861 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4921 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4981 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5041 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5101 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5161 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5221 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5281 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5341 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5401 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5461 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5521 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5581 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5641 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5701 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc ag
//
