LOCUS       sequence-329030-        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(C450A)-ShadowG
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        359..562
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     513..533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(607..609,610..612,613..1323)
                     /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(3163..3165,3166..3168,3169..3681)
                     /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    3992..4113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4029..4048)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4083..4102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4120..4575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4207..4226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4417..4438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4631..4735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4736..4804,4805..5596)
                     /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(4954..4973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5767..6355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6256..6275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
       61 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      541 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
      601 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      661 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      721 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      781 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
      841 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      901 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      961 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1021 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1081 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1141 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1201 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
     1261 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1321 aagtccggac tcagatctcg agctatggct accatcacct gcacccgatt cacggaagag
     1381 taccagctct tcgaggaact gggaaaggga gccttctccg tggtgcgcag gtgtgtgaag
     1441 gtgctggctg gccaggagta tgctgccaag attatcaaca ccaagaagct ctcagccaga
     1501 gatcaccaga agttggaacg cgaggcccgc atctgccgct tgttgaagca ccccaatatc
     1561 gtccgactcc atgacagcat ctccgaggag gggcaccact accttatctt cgatctggtc
     1621 actggtgggg agctgttcga agacattgtg gcccgggagt attacagtga ggctgatgcc
     1681 agccactgta tccagcagat cctggaggct gtgctacact gtcaccagat gggggtggtg
     1741 catcgcgacc tgaagcctga gaatctgttg ctggcttcga agctcaaggg tgctgcggtg
     1801 aagctggcag actttggcct ggccatagag gttgagggag agcagcaggc atggtttggg
     1861 ttcgcaggga cacctggata cctctcccca gaagtgctgc ggaaggaccc atacgggaag
     1921 cctgtggacc tgtgggcctg tggcgtcatc ctgtatatct tgctggttgg gtatccccca
     1981 ttctgggatg aggaccagca ccgcctgtac cagcagatca aagctggtgc ctacgatttc
     2041 ccatcaccag aatgggacac cgtcaccccg gaagccaagg atctgatcaa taagatgctg
     2101 accatcaacc cgtccaaacg catcacggcc gctgaggctc tcaagcaccc ctggatctcg
     2161 caccgctcca agcttctcga acgcattgaa aaaaactttg tgattaccga tccgcgcctg
     2221 ccggataacc cgattatttt tgcgagcgat agctttctgc agctgaccga atatagccgc
     2281 gaagaaattc tgggccgcaa cgcccgcttt ctgcagggcc cggaaaccga tcgcgcgacc
     2341 gtgcgcaaaa ttcgcgatgc gattgataac cagaccgaag tgaccgtgca gctgattaac
     2401 tataccaaaa gcggcaaaaa attttggaac ctgtttcatc tgcagccgat gcgcgatcag
     2461 aaaggcgatg tgcagtattt tattggcgtg cagctggatg gcaccgaaca tgtgcgcgat
     2521 gcggcggaac gcgaaggcgt gatgctgatt aaaaaaaccg cggaaaacat tgatgaagcg
     2581 gcgaaagagc tcgaattcgc caagtgcatg cacagacagg agaccgtgga ctgcctgaag
     2641 aagttcaatg ccaggaggaa actgaaggga gccatcctca ccactatgct ggccaccagg
     2701 aacttctccg tgcgcaaaca ggaaattatc aaagtgacag agcagctgat cgaagccata
     2761 agcaatggag actttgaatc ctacacgaag atgtgcgacc ctggaatgac agcctttgaa
     2821 ccggaggccc tggggaacct ggtcgagggc ctggaccttc atcgattcta ttttgaaaac
     2881 ctgtggtccc ggaacagcaa gcccgtgcac accaccatcc tgaaccctca cgtccacctg
     2941 atgggtgacg agtcagcctg catcacctac atccgcacca ctcagtacct ggatgcgggt
     3001 ggcatccccc gcacggccca gtcagaggag acccgtgtct ggcaccgcag ggatggaaaa
     3061 tggcagatcg tccacttcca cagatcaggg gcgccctccg tcctgcccca tggatccaac
     3121 cagcagatat tcctgcggga catagaacag gtgccacagc agatggtgag caagggcgag
     3181 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3241 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3301 ctcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
     3361 tacggcctga tgtgcttcgc gcgctatcca gaccacatga agcagcacga cttcttcaag
     3421 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3481 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3541 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctcga gtacaactgg
     3601 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     3661 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3721 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3781 aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3841 gccgccggga tcactctcgg catggactaa gcggccgcga cactagatca taatcagcca
     3901 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     3961 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     4021 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4081 ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta cgcgccctgt agcggcgcat
     4141 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     4201 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4261 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4321 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4381 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4441 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     4501 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4561 taacgcttac aatttcctct agataataat ggtttcttag acgtcaggtg gcacttttcg
     4621 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4681 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4741 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4801 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4861 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4921 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4981 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5041 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5101 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5161 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5221 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5281 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5341 gcaacaactg atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5401 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5461 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5521 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5581 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5641 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5701 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5761 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5821 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5881 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5941 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6001 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6061 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6121 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6181 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6241 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6301 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6361 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatggctcg
     6421 acagatcccc tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcattagt
     6481 ta
//
