LOCUS       sequence-329025-        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(K42M)-ShadowG
FEATURES             Location/Qualifiers
     source          1..6482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        65..368
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        369..572
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     523..543
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      611..620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(617..619,620..622,623..1333)
                     /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(3173..3175,3176..3178,3179..3691)
                     /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    4002..4123
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4039..4058)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4093..4112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4130..4585
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4217..4236)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4427..4448
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4641..4745
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4746..4814,4815..5606)
                     /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(4964..4983)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5777..6365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6266..6285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
       61 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
      121 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
      181 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
      241 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
      301 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      361 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      421 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      481 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      541 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      601 gctaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      661 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      721 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      781 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      841 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      901 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      961 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1021 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1081 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1141 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1201 gctgcccgac aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa
     1261 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1321 cgagctgtac aagtccggac tcagatctcg agctatggct accatcacct gcacccgatt
     1381 cacggaagag taccagctct tcgaggaact gggaaaggga gccttctccg tggtgcgcag
     1441 gtgtgtgaag gtgctggctg gccaggagta tgctgccatg attatcaaca ccaagaagct
     1501 ctcagccaga gatcaccaga agttggaacg cgaggcccgc atctgccgct tgttgaagca
     1561 ccccaatatc gtccgactcc atgacagcat ctccgaggag gggcaccact accttatctt
     1621 cgatctggtc actggtgggg agctgttcga agacattgtg gcccgggagt attacagtga
     1681 ggctgatgcc agccactgta tccagcagat cctggaggct gtgctacact gtcaccagat
     1741 gggggtggtg catcgcgacc tgaagcctga gaatctgttg ctggcttcga agctcaaggg
     1801 tgctgcggtg aagctggcag actttggcct ggccatagag gttgagggag agcagcaggc
     1861 atggtttggg ttcgcaggga cacctggata cctctcccca gaagtgctgc ggaaggaccc
     1921 atacgggaag cctgtggacc tgtgggcctg tggcgtcatc ctgtatatct tgctggttgg
     1981 gtatccccca ttctgggatg aggaccagca ccgcctgtac cagcagatca aagctggtgc
     2041 ctacgatttc ccatcaccag aatgggacac cgtcaccccg gaagccaagg atctgatcaa
     2101 taagatgctg accatcaacc cgtccaaacg catcacggcc gctgaggctc tcaagcaccc
     2161 ctggatctcg caccgctcca agcttctgga acgcattgaa aaaaactttg tgattaccga
     2221 tccgcgcctg ccggataacc cgattatttt tgcgagcgat agctttctgc agctgaccga
     2281 atatagccgc gaagaaattc tgggccgcaa ctgccgcttt ctgcagggcc cggaaaccga
     2341 tcgcgcgacc gtgcgcaaaa ttcgcgatgc gattgataac cagaccgaag tgaccgtgca
     2401 gctgattaac tataccaaaa gcggcaaaaa attttggaac ctgtttcatc tgcagccgat
     2461 gcgcgatcag aaaggcgatg tgcagtattt tattggcgtg cagctggatg gcaccgaaca
     2521 tgtgcgcgat gcggcggaac gcgaaggcgt gatgctgatt aaaaaaaccg cggaaaacat
     2581 tgatgaagcg gcgaaagagc tcgaattcgc caagtgcatg cacagacagg agaccgtgga
     2641 ctgcctgaag aagttcaatg ccaggaggaa actgaaggga gccatcctca ccactatgct
     2701 ggccaccagg aacttctccg tgcgcaaaca ggaaattatc aaagtgacag agcagctgat
     2761 cgaagccata agcaatggag actttgaatc ctacacgaag atgtgcgacc ctggaatgac
     2821 agcctttgaa ccggaggccc tggggaacct ggtcgagggc ctggaccttc atcgattcta
     2881 ttttgaaaac ctgtggtccc ggaacagcaa gcccgtgcac accaccatcc tgaaccctca
     2941 cgtccacctg atgggtgacg agtcagcctg catcacctac atccgcacca ctcagtacct
     3001 ggatgcgggt ggcatccccc gcacggccca gtcagaggag acccgtgtct ggcaccgcag
     3061 ggatggaaaa tggcagatcg tccacttcca cagatcaggg gcgccctccg tcctgcccca
     3121 tggatccaac cagcagatat tcctgcggga catagaacag gtgccacagc agatggtgag
     3181 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     3241 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     3301 gaccctgaag ctcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     3361 caccttcggc tacggcctga tgtgcttcgc gcgctatcca gaccacatga agcagcacga
     3421 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     3481 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     3541 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctcga
     3601 gtacaactgg aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     3661 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     3721 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     3781 cacccagtcc aagctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     3841 gttcgtgacc gccgccggga tcactctcgg catggactaa gcggccgcga cactagatca
     3901 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3961 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     4021 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4081 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaacgcgta cgcgccctgt
     4141 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     4201 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     4261 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     4321 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     4381 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     4441 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     4501 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     4561 aacaaaatat taacgcttac aatttcctct agataataat ggtttcttag acgtcaggtg
     4621 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4681 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4741 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     4801 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     4861 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     4921 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     4981 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5041 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5101 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5161 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5221 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5281 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5341 tagcttcccg gcaacaactg atagactgga tggaggcgga taaagttgca ggaccacttc
     5401 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5461 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5521 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5581 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     5641 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     5701 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     5761 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     5821 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     5881 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     5941 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6001 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6061 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6121 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6181 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6241 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6301 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6361 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6421 acatggctcg acagatcccc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6481 at
//
