LOCUS       sequence-329024-        6482 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses paCaMKII in mammalian cells to measure structural change 
            of paCaMKII..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6482)
  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 6482)
  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: CMV-mEGFP(A206K)-paCaMKII(T286A)-ShadowG
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        43..346
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        347..550
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     501..521
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      589..598
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(595..597,598..600,601..1311)
                     /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"
     CDS             join(3151..3153,3154..3156,3157..3669)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     polyA_signal    3980..4101
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4017..4036)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4071..4090
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4108..4563
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4195..4214)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4405..4426
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4619..4723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4724..4792,4793..5584)
                     /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(4942..4961)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5755..6343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6244..6263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
       61 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      121 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      181 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      241 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      301 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      361 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      421 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      481 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      541 aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg
      601 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      661 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      721 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      781 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      841 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      901 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
      961 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1021 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1081 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1141 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1201 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1261 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc
     1321 agatctcgag ctatggctac catcacctgc acccgattca cggaagagta ccagctcttc
     1381 gaggaactgg gaaagggagc cttctccgtg gtgcgcaggt gtgtgaaggt gctggctggc
     1441 caggagtatg ctgccaagat tatcaacacc aagaagctct cagccagaga tcaccagaag
     1501 ttggaacgcg aggcccgcat ctgccgcttg ttgaagcacc ccaatatcgt ccgactccat
     1561 gacagcatct ccgaggaggg gcaccactac cttatcttcg atctggtcac tggtggggag
     1621 ctgttcgaag acattgtggc ccgggagtat tacagtgagg ctgatgccag ccactgtatc
     1681 cagcagatcc tggaggctgt gctacactgt caccagatgg gggtggtgca tcgcgacctg
     1741 aagcctgaga atctgttgct ggcttcgaag ctcaagggtg ctgcggtgaa gctggcagac
     1801 tttggcctgg ccatagaggt tgagggagag cagcaggcat ggtttgggtt cgcagggaca
     1861 cctggatacc tctccccaga agtgctgcgg aaggacccat acgggaagcc tgtggacctg
     1921 tgggcctgtg gcgtcatcct gtatatcttg ctggttgggt atcccccatt ctgggatgag
     1981 gaccagcacc gcctgtacca gcagatcaaa gctggtgcct acgatttccc atcaccagaa
     2041 tgggacaccg tcaccccgga agccaaggat ctgatcaata agatgctgac catcaacccg
     2101 tccaaacgca tcacggccgc tgaggctctc aagcacccct ggatctcgca ccgctccaag
     2161 cttctggaac gcattgaaaa aaactttgtg attaccgatc cgcgcctgcc ggataacccg
     2221 attatttttg cgagcgatag ctttctgcag ctgaccgaat atagccgcga agaaattctg
     2281 ggccgcaact gccgctttct gcagggcccg gaaaccgatc gcgcgaccgt gcgcaaaatt
     2341 cgcgatgcga ttgataacca gaccgaagtg accgtgcagc tgattaacta taccaaaagc
     2401 ggcaaaaaat tttggaacct gtttcatctg cagccgatgc gcgatcagaa aggcgatgtg
     2461 cagtatttta ttggcgtgca gctggatggc accgaacatg tgcgcgatgc ggcggaacgc
     2521 gaaggcgtga tgctgattaa aaaaaccgcg gaaaacattg atgaagcggc gaaagagctc
     2581 gaattcgcca agtgcatgca cagacaggag gccgtggact gcctgaagaa gttcaatgcc
     2641 aggaggaaac tgaagggagc catcctcacc actatgctgg ccaccaggaa cttctccgtg
     2701 cgcaaacagg aaattatcaa agtgacagag cagctgatcg aagccataag caatggagac
     2761 tttgaatcct acacgaagat gtgcgaccct ggaatgacag cctttgaacc ggaggccctg
     2821 gggaacctgg tcgagggcct ggaccttcat cgattctatt ttgaaaacct gtggtcccgg
     2881 aacagcaagc ccgtgcacac caccatcctg aaccctcacg tccacctgat gggtgacgag
     2941 tcagcctgca tcacctacat ccgcaccact cagtacctgg atgcgggtgg catcccccgc
     3001 acggcccagt cagaggagac ccgtgtctgg caccgcaggg atggaaaatg gcagatcgtc
     3061 cacttccaca gatcaggggc gccctccgtc ctgccccatg gatccaacca gcagatattc
     3121 ctgcgggaca tagaacaggt gccacagcag atggtgagca agggcgagga gctgttcacc
     3181 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3241 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct catctgcacc
     3301 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgatg
     3361 tgcttcgcgc gctatccaga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3421 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3481 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3541 ttcaaggagg acggcaacat cctggggcac aagctcgagt acaactggaa cagccacaac
     3601 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3661 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3721 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     3781 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3841 actctcggca tggactaagc ggccgcgaca ctagatcata atcagccata ccacatttgt
     3901 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     3961 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4021 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4081 caaactcatc aatgtatctt aacgcgtacg cgccctgtag cggcgcatta agcgcggcgg
     4141 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4201 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4261 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4321 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4381 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4441 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4501 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4561 tttcctctag ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4621 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4681 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4741 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4801 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4861 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4921 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4981 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5041 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5101 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5161 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5221 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5281 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaactgat
     5341 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5401 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5461 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5521 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5581 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5641 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5701 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5761 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5821 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5881 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5941 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6001 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6061 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6121 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6181 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6241 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6301 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6361 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac agatcccctg
     6421 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta ttaatagtaa
     6481 tc
//
