LOCUS       sequence-329026-        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(I539E)-ShadowG
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        103..406
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        407..610
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     561..581
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      649..658
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(655..657,658..660,661..1371)
                     /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(3211..3213,3214..3216,3217..3729)
                     /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    4040..4161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4077..4096)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4131..4150
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4168..4623
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4255..4274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4465..4486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4679..4783
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4784..4852,4853..5644)
                     /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(5002..5021)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5815..6403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6304..6323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatcccctg attctgtgga taaccgtatt accgccatgc attagttatt aatagtaatc
       61 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
      121 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
      181 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
      241 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
      301 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
      361 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
      421 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
      481 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
      541 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
      601 aagcagagct ggtttagtga accgtcagat ccgctagcgc taccggtcgc caccatggtg
      661 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      721 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag
      781 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
      841 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac
      901 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag
      961 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac
     1021 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg
     1081 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc
     1141 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac
     1201 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1261 agcacccagt ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg
     1321 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtccggactc
     1381 agatctcgag ctatggctac catcacctgc acccgattca cggaagagta ccagctcttc
     1441 gaggaactgg gaaagggagc cttctccgtg gtgcgcaggt gtgtgaaggt gctggctggc
     1501 caggagtatg ctgccaagat tatcaacacc aagaagctct cagccagaga tcaccagaag
     1561 ttggaacgcg aggcccgcat ctgccgcttg ttgaagcacc ccaatatcgt ccgactccat
     1621 gacagcatct ccgaggaggg gcaccactac cttatcttcg atctggtcac tggtggggag
     1681 ctgttcgaag acattgtggc ccgggagtat tacagtgagg ctgatgccag ccactgtatc
     1741 cagcagatcc tggaggctgt gctacactgt caccagatgg gggtggtgca tcgcgacctg
     1801 aagcctgaga atctgttgct ggcttcgaag ctcaagggtg ctgcggtgaa gctggcagac
     1861 tttggcctgg ccatagaggt tgagggagag cagcaggcat ggtttgggtt cgcagggaca
     1921 cctggatacc tctccccaga agtgctgcgg aaggacccat acgggaagcc tgtggacctg
     1981 tgggcctgtg gcgtcatcct gtatatcttg ctggttgggt atcccccatt ctgggatgag
     2041 gaccagcacc gcctgtacca gcagatcaaa gctggtgcct acgatttccc atcaccagaa
     2101 tgggacaccg tcaccccgga agccaaggat ctgatcaata agatgctgac catcaacccg
     2161 tccaaacgca tcacggccgc tgaggctctc aagcacccct ggatctcgca ccgctccaag
     2221 cttctggaac gcattgaaaa aaactttgtg attaccgatc cgcgcctgcc ggataacccg
     2281 attatttttg cgagcgatag ctttctgcag ctgaccgaat atagccgcga agaaattctg
     2341 ggccgcaact gccgctttct gcagggcccg gaaaccgatc gcgcgaccgt gcgcaaaatt
     2401 cgcgatgcga ttgataacca gaccgaagtg accgtgcagc tgattaacta taccaaaagc
     2461 ggcaaaaaat tttggaacct gtttcatctg cagccgatgc gcgatcagaa aggcgatgtg
     2521 cagtatttta ttggcgtgca gctggatggc accgaacatg tgcgcgatgc ggcggaacgc
     2581 gaaggcgtga tgctgattaa aaaaaccgcg gaaaacgaag atgaagcggc gaaagagctc
     2641 gaattcgcca agtgcatgca cagacaggag accgtggact gcctgaagaa gttcaatgcc
     2701 aggaggaaac tgaagggagc catcctcacc actatgctgg ccaccaggaa cttctccgtg
     2761 cgcaaacagg aaattatcaa agtgacagag cagctgatcg aagccataag caatggagac
     2821 tttgaatcct acacgaagat gtgcgaccct ggaatgacag cctttgaacc ggaggccctg
     2881 gggaacctgg tcgagggcct ggaccttcat cgattctatt ttgaaaacct gtggtcccgg
     2941 aacagcaagc ccgtgcacac caccatcctg aaccctcacg tccacctgat gggtgacgag
     3001 tcagcctgca tcacctacat ccgcaccact cagtacctgg atgcgggtgg catcccccgc
     3061 acggcccagt cagaggagac ccgtgtctgg caccgcaggg atggaaaatg gcagatcgtc
     3121 cacttccaca gatcaggggc gccctccgtc ctgccccatg gatccaacca gcagatattc
     3181 ctgcgggaca tagaacaggt gccacagcag atggtgagca agggcgagga gctgttcacc
     3241 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3301 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagct catctgcacc
     3361 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccttcggcta cggcctgatg
     3421 tgcttcgcgc gctatccaga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3481 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3541 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3601 ttcaaggagg acggcaacat cctggggcac aagctcgagt acaactggaa cagccacaac
     3661 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     3721 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     3781 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa
     3841 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     3901 actctcggca tggactaagc ggccgcgaca ctagatcata atcagccata ccacatttgt
     3961 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     4021 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     4081 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4141 caaactcatc aatgtatctt aacgcgtacg cgccctgtag cggcgcatta agcgcggcgg
     4201 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     4261 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     4321 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4381 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4441 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     4501 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     4561 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     4621 tttcctctag ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     4681 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4741 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     4801 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     4861 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4921 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4981 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     5041 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     5101 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     5161 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     5221 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     5281 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     5341 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaactgat
     5401 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     5461 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     5521 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     5581 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     5641 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     5701 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5761 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5821 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     5881 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5941 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     6001 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6061 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6121 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6181 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6241 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6301 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6361 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6421 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atggctcgac agatcccctg
     6481 cg
//
