LOCUS       sequence-325945-        5716 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses light-insensitive photoactivatable CaMKII(SD: Super Dark) 
            in mammalian cells as a negative control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5716)
  AUTHORS   Shibata ACE, Ueda HH, Eto K, Onda M, Sato A, Ohba T, Nabekura J, 
            Murakoshi H
  TITLE     Photoactivatable CaMKII induces synaptic plasticity in single 
            synapses.
  JOURNAL   Nat Commun. 2021 Feb 2;12(1):751. doi: 10.1038/s41467-021-21025-6.
  PUBMED    33531495
REFERENCE   2  (bases 1 to 5716)
  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: "2021-02-2"; volume: "12(1):751. doi"; pages: " 
            10.1038/s41467-021-21025-6"
COMMENT     Sequence Label: pAAV-CaMP0.4-FHS-paCaMKII(SD)-WPRE3
FEATURES             Location/Qualifiers
     source          1..5716
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      584..593
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             593..616
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             617..634
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             638..661
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             join(689..697,698..712)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(2599..2619)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   2979..3119
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2979..3108
                     /label=AAV2 ITR
     rep_origin      3194..3649
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3281..3300)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3491..3512
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3666..3685)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3785..3807
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3845..3863)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3931..4035
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4036..4104,4105..4896)
                     /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(4254..4273)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5067..5655
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5556..5575
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtctgtta aaagggccct atagttggag gtgggggagg
      181 taggaagagc gatgatcact tgtggactaa gtttgttcgc atccccttct ccaaccccct
      241 cagtacatca ccctggggga acagggtcca cttgctcctg ggcccacaca gtcctgcagt
      301 attgtgtata taaggccagg gcaaagagga gcaggtttta aagtgaaagg caggcaggtg
      361 ttggggaggc agttaccggg gcaacgggaa cagggcgttt cggaggtggt tgccatgggg
      421 acctggatgc tgacgaaggc tcgcgaggct gtgagcagcc acagtgccct gctcagaagc
      481 cccaagctcg tcagtcaagc cggttctccg tttgcactca ggagcacggg caggcgagtg
      541 gcccctagtt ctgggggcag cgggggatct ccgggtaccg gtcgccacca tggattacaa
      601 ggacgatgac gataagcatc atcatcatca tcataagtgg tcacatccac aattcgagaa
      661 gggtactggt ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctgta
      721 caagtccgga ctcagatctc gagctatggc taccatcacc tgcacccgat tcacggaaga
      781 gtaccagctc ttcgaggaac tgggaaaggg agccttctcc gtggtgcgca ggtgtgtgaa
      841 ggtgctggct ggccaggagt atgctgccaa gattatcaac accaagaagc tctcagccag
      901 agatcaccag aagttggaac gcgaggcccg catctgccgc ttgttgaagc accccaatat
      961 cgtccgactc catgacagca tctccgagga ggggcaccac taccttatct tcgatctggt
     1021 cactggtggg gagctgttcg aagacattgt ggcccgggag tattacagtg aggctgatgc
     1081 cagccactgt atccagcaga tcctggaggc tgtgctacac tgtcaccaga tgggggtggt
     1141 gcatcgcgac ctgaagcctg agaatctgtt gctggcttcg aagctcaagg gtgctgcggt
     1201 gaagctggca gactttggcc tggccataga ggttgaggga gagcagcagg catggtttgg
     1261 gttcgcaggg acacctggat acctctcccc agaagtgctg cggaaggacc catacgggaa
     1321 gcctgtggac ctgtgggcct gtggcgtcat cctgtatatc ttgctggttg ggtatccccc
     1381 attctgggat gaggaccagc accgcctgta ccagcagatc aaagctggtg cctacgattt
     1441 cccatcacca gaatgggaca ccgtcacccc ggaagccaag gatctgatca ataagatgct
     1501 gaccatcaac ccgtccaaac gcatcacggc cgctgaggct ctcaagcacc cctggatctc
     1561 gcaccgctca aagcttcttg aacgtattga gaagaacttt gtcattactg acccaagatt
     1621 gccagataat cccattatat tcgcgtccga tagtttcttg cagttgacag aatatagccg
     1681 tgaagaaatt ttgggaagaa acgccaggtt tctacaaggt cctgaaactg atcgcgcgac
     1741 agtgagaaaa attagagatg ccatagataa ccaaacagag gtcactgttc agctgattaa
     1801 ttatacaaag agtggtaaaa agttctggaa cctctttcac ttgcagccta tgcgagatca
     1861 gaagggagat gtccagtact ttattggggt tcagaaggat ggaactgagc atgtccgaga
     1921 tgctgccgag agagaggcag tcatggagat taagaaaact gcagaagaaa ttgatgaggc
     1981 ggcaaaagag ctcgaattcg ccaagtgcat gcacagacag gagaccgtgg actgcctgaa
     2041 gaagttcaat gccaggagga aactgaaggg agccatcctc accactatgc tggccaccag
     2101 gaacttctcc gtgcgcaaac aggaaattat caaagtgaca gagcagctga tcgaagccat
     2161 aagcaatgga gactttgaat cctacacgaa gatgtgcgac cctggaatga cagcctttga
     2221 accggaggcc ctggggaacc tggtcgaggg cctggacctt catcgattct attttgaaaa
     2281 cctgtggtcc cggaacagca agcccgtgca caccaccatc ctgaaccctc acgtccacct
     2341 gatgggtgac gagtcagcct gcatcaccta catccgcacc actcagtacc tggatgcggg
     2401 tggcatcccc cgcacggccc agtcagagga gacccgtgtc tggcaccgca gggatggaaa
     2461 atggcagatc gtccacttcc acagatcagg ggcgccctcc gtcctgcccc atggatccta
     2521 agcggccgcc aattccgctc gagataatca acctctggat tacaaaattt gtgaaagatt
     2581 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     2641 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     2701 gttagttctt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
     2761 tcggctgttg ggcactgaca attccgtggt gtttatttgt gaaatttgtg atgctattgc
     2821 tttatttgta accatctagc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     2881 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     2941 ttcaggggga gatgtgggag gttttttaaa gcggccgcag gaacccctag tgatggagtt
     3001 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3061 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     3121 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     3181 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3241 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     3301 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3361 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     3421 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3481 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     3541 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     3601 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     3661 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     3721 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3781 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     3841 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3901 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3961 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4021 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4081 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4141 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4201 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4261 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4321 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4381 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4441 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4501 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4561 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4621 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4681 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4741 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4801 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4861 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4921 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4981 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5041 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5101 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5161 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5221 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5281 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5341 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5401 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5461 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5521 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5581 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5641 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5701 ccttttgctc acatgt
//
