LOCUS       sequence-329023-        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-ShadowG
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        102..405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..609
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     560..580
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      648..657
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(654..656,657..659,660..1370)
                     /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(3210..3212,3213..3215,3216..3728)
                     /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    4039..4160
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4076..4095)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4130..4149
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4167..4622
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4254..4273)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4464..4485
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4678..4782
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4783..4851,4852..5643)
                     /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(5001..5020)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5814..6402
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6303..6322
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
       61 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      421 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      541 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      601 agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc accatggtga
      661 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      721 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      781 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
      841 ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg
      901 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      961 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1021 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1081 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     1141 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     1201 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     1261 gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     1321 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag tccggactca
     1381 gatctcgagc tatggctacc atcacctgca cccgattcac ggaagagtac cagctcttcg
     1441 aggaactggg aaagggagcc ttctccgtgg tgcgcaggtg tgtgaaggtg ctggctggcc
     1501 aggagtatgc tgccaagatt atcaacacca agaagctctc agccagagat caccagaagt
     1561 tggaacgcga ggcccgcatc tgccgcttgt tgaagcaccc caatatcgtc cgactccatg
     1621 acagcatctc cgaggagggg caccactacc ttatcttcga tctggtcact ggtggggagc
     1681 tgttcgaaga cattgtggcc cgggagtatt acagtgaggc tgatgccagc cactgtatcc
     1741 agcagatcct ggaggctgtg ctacactgtc accagatggg ggtggtgcat cgcgacctga
     1801 agcctgagaa tctgttgctg gcttcgaagc tcaagggtgc tgcggtgaag ctggcagact
     1861 ttggcctggc catagaggtt gagggagagc agcaggcatg gtttgggttc gcagggacac
     1921 ctggatacct ctccccagaa gtgctgcgga aggacccata cgggaagcct gtggacctgt
     1981 gggcctgtgg cgtcatcctg tatatcttgc tggttgggta tcccccattc tgggatgagg
     2041 accagcaccg cctgtaccag cagatcaaag ctggtgccta cgatttccca tcaccagaat
     2101 gggacaccgt caccccggaa gccaaggatc tgatcaataa gatgctgacc atcaacccgt
     2161 ccaaacgcat cacggccgct gaggctctca agcacccctg gatctcgcac cgctccaagc
     2221 ttctggaacg cattgaaaaa aactttgtga ttaccgatcc gcgcctgccg gataacccga
     2281 ttatttttgc gagcgatagc tttctgcagc tgaccgaata tagccgcgaa gaaattctgg
     2341 gccgcaactg ccgctttctg cagggcccgg aaaccgatcg cgcgaccgtg cgcaaaattc
     2401 gcgatgcgat tgataaccag accgaagtga ccgtgcagct gattaactat accaaaagcg
     2461 gcaaaaaatt ttggaacctg tttcatctgc agccgatgcg cgatcagaaa ggcgatgtgc
     2521 agtattttat tggcgtgcag ctggatggca ccgaacatgt gcgcgatgcg gcggaacgcg
     2581 aaggcgtgat gctgattaaa aaaaccgcgg aaaacattga tgaagcggcg aaagagctcg
     2641 aattcgccaa gtgcatgcac agacaggaga ccgtggactg cctgaagaag ttcaatgcca
     2701 ggaggaaact gaagggagcc atcctcacca ctatgctggc caccaggaac ttctccgtgc
     2761 gcaaacagga aattatcaaa gtgacagagc agctgatcga agccataagc aatggagact
     2821 ttgaatccta cacgaagatg tgcgaccctg gaatgacagc ctttgaaccg gaggccctgg
     2881 ggaacctggt cgagggcctg gaccttcatc gattctattt tgaaaacctg tggtcccgga
     2941 acagcaagcc cgtgcacacc accatcctga accctcacgt ccacctgatg ggtgacgagt
     3001 cagcctgcat cacctacatc cgcaccactc agtacctgga tgcgggtggc atcccccgca
     3061 cggcccagtc agaggagacc cgtgtctggc accgcaggga tggaaaatgg cagatcgtcc
     3121 acttccacag atcaggggcg ccctccgtcc tgccccatgg atccaaccag cagatattcc
     3181 tgcgggacat agaacaggtg ccacagcaga tggtgagcaa gggcgaggag ctgttcaccg
     3241 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3301 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagctc atctgcacca
     3361 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cttcggctac ggcctgatgt
     3421 gcttcgcgcg ctatccagac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3481 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3541 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3601 tcaaggagga cggcaacatc ctggggcaca agctcgagta caactggaac agccacaacg
     3661 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     3721 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     3781 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccaag ctgagcaaag
     3841 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     3901 ctctcggcat ggactaagcg gccgcgacac tagatcataa tcagccatac cacatttgta
     3961 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     4021 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4081 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4141 aaactcatca atgtatctta acgcgtacgc gccctgtagc ggcgcattaa gcgcggcggg
     4201 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     4261 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     4321 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     4381 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     4441 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     4501 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4561 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     4621 ttcctctaga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4681 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4741 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     4801 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     4861 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     4921 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     4981 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5041 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5101 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5161 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5221 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5281 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5341 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaactgata
     5401 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5461 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5521 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5581 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5641 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5701 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5761 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5821 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5881 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5941 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6001 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6061 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6121 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6181 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6241 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6301 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6361 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6421 tttttacggt tcctggcctt ttgctggcct tttgctcaca tggctcgaca gatcccctgc
     6481 gt
//
