LOCUS       sequence-434512-        8416 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Improved FLIM sensor for CaMK2-alpha activation (mammalian 
            expression, cytosol).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8416)
  AUTHORS   Jain A, Nakahata Y, Pancani T, Watabe T, Rusina P, South K, Adachi 
            K, Yan L, Simorowski N, Furukawa H, Yasuda R
  TITLE     Dendritic, delayed, stochastic CaMKII activation in behavioural time
            scale plasticity.
  JOURNAL   Nature. 2024 Oct 9. doi: 10.1038/s41586-024-08021-8.
  PUBMED    39385027
REFERENCE   2  (bases 1 to 8416)
  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: "Nature. 
            2024 Oct 9. doi: 10.1038/s41586-024-08021-8."
COMMENT     Sequence Label: pCAG-2dV-Camui (wt)
FEATURES             Location/Qualifiers
     source          1..8416
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..668
                     /label=chicken beta-actin promoter
     intron          669..1689
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1609..1631
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1697..1716
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1760..1769
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1766..1768,1769..1771,1772..2482)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2501..2510
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2507..2509,2510..2512,2513..3223)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNWNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      4697..4706
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4703..4705,4706..4708,4709..5422)
                     /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"
     primer_bind     complement(5438..5457)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    5503..5558
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(5557..5576)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5919..5935)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5919..5935)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5932..5954)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5943..5959
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5967..5973,5974..5991,5992..5997))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6012..6033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6092..6287
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      6138..6273
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    6293..6427
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6330..6349)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6384..6403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6495..6512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6666..7254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6746..6765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7425..8216,8217..8285))
                     /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     8048..8067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8286..8390)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggg ggcgcgcgcc rggsggggsg gggsggggsg
      541 rggggsgggg sggggcgrgg cggagaggtg cggcggcagc caatcagagc ggcgcgctcc
      601 gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc
      661 ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc
      721 gccgcccgcc ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc
      781 cttctcctcc gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc
      841 tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggg
      901 gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg ctccgcgctg cccggcggct
      961 gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg cagtgtgcgc gaggggagcg
     1021 cggccggggg cggtgccccg cggtgcgggg ggggctgcga ggggaacaaa ggctgcgtgc
     1081 ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc gtcggtcggg ctgcaacccc
     1141 ccctgcaccc ccctccccga gttgctgagc acggcccggc ttcgggtgcg gggctccgta
     1201 cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg gcggcaggtg ggggtgccgg
     1261 gcggggcggg gccgcctcgg gccggggrgg gctcggggga ggggcgcggc ggcccccgga
     1321 gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg ccttttatgg taatcgtgcg
     1381 agagggcgca gggacttcct ttgtcccaaa tctgtgcgga gccgaaatct gggaggcgcc
     1441 gccgcacccc ctctagcggg cgcggggcga agcggtgcgg cgccggcagg aaggaaatgg
     1501 gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct tctccctctc cagcctcggg
     1561 gctgtccgcg gggggacggc tgccttcggg ggggacgggg cagggcgggg ttcggcttct
     1621 ggcgtgtgac cggcggctct agctagagcc tctgctaacc atgttcatgc cttcttcttt
     1681 ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa
     1741 ttcgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt
     1801 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1861 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     1921 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt
     1981 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     2041 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     2101 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     2161 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tggaacagcc acaacgtcta
     2221 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     2281 cgaggacggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg
     2341 ccccgtgctg ctgcccgaca accactacct gagctaccag tccaagctga gcaaagaccc
     2401 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct
     2461 cggcatggac gagctgtaca agtccggagc gctacaggtc gccaccatgg tgagcaaggg
     2521 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2581 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2641 gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     2701 gggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     2761 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2821 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2881 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2941 ctggaacagc cacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     3001 cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgccgacc actaccagca
     3061 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     3121 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3181 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtctggtc tcagatctcg
     3241 agctatggct accatcacct gcacccgatt cacggaagag taccagctct tcgaggaact
     3301 gggaaaggga gccttctccg tggtgcgcag gtgtgtgaag gtgctggctg gccaggagta
     3361 tgctgccaag attatcaaca ccaagaagct ctcagccaga gatcaccaga agttggaacg
     3421 cgaggcccgc atctgccgct tgttgaagca ccccaatatc gtccgactcc atgacagcat
     3481 ctccgaggag gggcaccact accttatctt cgatctggtc actggtgggg agctgttcga
     3541 agacattgtg gcccgggagt attacagtga ggctgatgcc agccactgta tccagcagat
     3601 cctggaggct gtgctacact gtcaccagat gggggtggtg catcgcgacc tgaagcctga
     3661 gaatctgttg ctggcttcga agctcaaggg tgctgcggtg aagctggcag actttggcct
     3721 ggccatagag gttgagggag agcagcaggc atggtttggg ttcgcaggga cacctggata
     3781 cctctcccca gaagtgctgc ggaaggaccc atacgggaag cctgtggacc tgtgggcctg
     3841 tggcgtcatc ctgtatatct tgctggttgg gtatccccca ttctgggatg aggaccagca
     3901 ccgcctgtac cagcagatca aagctggtgc ctacgatttc ccatcaccag aatgggacac
     3961 cgtcaccccg gaagccaagg atctgatcaa taagatgctg accatcaacc cgtccaaacg
     4021 catcacggcc gctgaggctc tcaagcaccc ctggatctcg caccgctcca ctgtggcctc
     4081 ctgcatgcac agacaggaga ccgtggactg cctgaagaag ttcaatgcca ggaggaaact
     4141 gaagggagcc atcctcacca ctatgctggc caccaggaac ttctccggag ggaagagtgg
     4201 aggaaacaag aagaatgatg gcgtgaagga atcctctgag agcaccaaca ccaccatcga
     4261 ggatgaagac accaaagtgc gcaaacagga aattatcaaa gtgacagagc agctgatcga
     4321 agccataagc aatggagact ttgaatccta cacgaagatg tgcgaccctg gaatgacagc
     4381 ctttgaaccg gaggccctgg ggaacctggt cgagggcctg gactttcatc gattctattt
     4441 tgaaaacctg tggtcccgga acagcaagcc cgtgcacacc accatcctga accctcacat
     4501 ccacctgatg ggtgacgagt cagcctgcat cgcctacatc cgcatcactc agtacctgga
     4561 tgcgggtggc atcccccgca cggcccagtc agaggagacc cgtgtctggc accgcaggga
     4621 tggaaaatgg cagatcgtcc acttccacag atctggggcg ccctccgtcc tgccccatgg
     4681 actagcgcta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
     4741 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     4801 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     4861 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     4921 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     4981 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     5041 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     5101 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     5161 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     5221 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     5281 cgtgctgctg cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa
     5341 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     5401 catggacgag ctgtacaagt agtctagagt cgacggatct ttttccctct gccaaaaatt
     5461 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     5521 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     5581 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     5641 tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg aaacagcccc
     5701 ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt ttttttatat
     5761 tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat gttttactag
     5821 ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt ctcttatgga
     5881 gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt tcctgtgtga
     5941 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     6001 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     6061 cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca accatagtcc
     6121 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6241 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt
     6301 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     6361 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     6421 ctggatccgc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     6481 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     6541 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     6601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     7021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     7081 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     7141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     7201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     7261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     7321 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     7381 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     7441 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     7501 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     7561 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     7621 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7681 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7741 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7801 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7861 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7921 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7981 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     8041 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     8101 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     8161 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     8221 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     8281 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     8341 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     8401 cgaaaagtgc cacctg
//
