LOCUS       sequence-469102-        8415 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  T305D/T306D phosphomimetic mutant of pCAG-2dV-Camui.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8415)
  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 8415)
  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 (T305D/T306D)
FEATURES             Location/Qualifiers
     source          1..8415
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    828..962
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(852..871)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     906..925
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(968..1163)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      982..1117
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1222..1243
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1258..1263,1264..1281,1282..1288)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1296..1312
                     /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)."
     primer_bind     1301..1323
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1320..1336
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1320..1336
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1679..1698
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1697..1752
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1798..1817
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(1833..2546,2547..2549,2550..2552))
                     /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"
     regulatory      2549..2558
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(join(4032..4742,4743..4745,4746..4748))
                     /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      4745..4754
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(join(4773..5483,5484..5486,5487..5489))
                     /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      5486..5495
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(5539..5558)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          5566..6586
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(5624..5646)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     promoter        complement(6587..6864)
                     /label=chicken beta-actin promoter
     enhancer        6866..7245
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7280..7384
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7385..7453,7454..8245)
                     /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(7603..7622)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagcggat ccagacatga
      841 taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta
      901 tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag
      961 ttagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc tggaatagct
     1021 cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag ccatggggcg
     1081 gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg ggcgggacta
     1141 tggttgctga ctaattgaga tgcggatccg ctggcacgac aggtttcccg actggaaagc
     1201 gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt
     1261 acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac
     1321 aggaaacagc tatgaccatg attacgccaa gcttgggctg caggtcgagg gatctccata
     1381 agagaagagg gacagctatg actgggagta gtcaggagag gaggaaaaat ctggctagta
     1441 aaacatgtaa ggaaaatttt agggatgtta aagaaaaaaa taacacaaaa caaaatataa
     1501 aaaaaatcta acctcaagtc aaggcttttc tatggaataa ggaatggaca gcagggggct
     1561 gtttcatata ctgatgacct ctttatagcc aacctttgtt catggcagcc agcatatggg
     1621 catatgttgc caaactctaa accaaatact cattctgatg ttttaaatga tttgccctcc
     1681 catatgtcct tccgagtgag agacacaaaa aattccaaca cactattgca atgaaaataa
     1741 atttccttta ttagccagaa gtcagatgct caaggggctt catgatgtcc ccataatttt
     1801 tggcagaggg aaaaagatcc gtcgactcta gactacttgt acagctcgtc catgccgaga
     1861 gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt ctcgttgggg
     1921 tctttgctca gcttggactg ggtgctcagg tagtggttgt cgggcagcag cacggggccg
     1981 tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgctgcc gtcctcgatg
     2041 ttgtggcgga tcttgaagtt caccttgatg ccgttcttct gcttgtcggc catgatatag
     2101 acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc gtcctccttg
     2161 aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa cttcacctcg
     2221 gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg gacgtagcct
     2281 tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta gcggctgaag
     2341 cactgcacgc cgtaggtcag ggtggtcacg agggtgggcc agggcacggg cagcttgccg
     2401 gtggtgcaga tgaacttcag ggtcagcttg ccgtaggtgg catcgccctc gccctcgccg
     2461 gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat gggcaccacc
     2521 ccggtgaaca gctcctcgcc cttgctcacc atggtggcga ccggtagcgc tagtccatgg
     2581 ggcaggacgg agggcgcccc agatctgtgg aagtggacga tctgccattt tccatccctg
     2641 cggtgccaga cacgggtctc ctctgactgg gccgtgcggg ggatgccacc cgcatccagg
     2701 tactgagtga tgcggatgta ggcgatgcag gctgactcgt cacccatcag gtggatgtga
     2761 gggttcagga tggtggtgtg cacgggcttg ctgttccggg accacaggtt ttcaaaatag
     2821 aatcgatgaa agtccaggcc ctcgaccagg ttccccaggg cctccggttc aaaggctgtc
     2881 attccagggt cgcacatctt cgtgtaggat tcaaagtctc cattgcttat ggcttcgatc
     2941 agctgctctg tcactttgat aatttcctgt ttgcgcactt tggtgtcttc atcctcgatg
     3001 gtggtgttgg tgctctcaga ggattccttc acgccatcat tcttcttgtt tcctccactc
     3061 ttccctccgg agaagttcct ggtggccagc atatcgtcga ggatggctcc cttcagtttc
     3121 ctcctggcat tgaacttctt caggcagtcc acggtctcct gtctgtgcat gcaggaggcc
     3181 acagtggagc ggtgcgagat ccaggggtgc ttgagagcct cagcggccgt gatgcgtttg
     3241 gacgggttga tggtcagcat cttattgatc agatccttgg cttccggggt gacggtgtcc
     3301 cattctggtg atgggaaatc gtaggcacca gctttgatct gctggtacag gcggtgctgg
     3361 tcctcatccc agaatggggg atacccaacc agcaagatat acaggatgac gccacaggcc
     3421 cacaggtcca caggcttccc gtatgggtcc ttccgcagca cttctgggga gaggtatcca
     3481 ggtgtccctg cgaacccaaa ccatgcctgc tgctctccct caacctctat ggccaggcca
     3541 aagtctgcca gcttcaccgc agcacccttg agcttcgaag ccagcaacag attctcaggc
     3601 ttcaggtcgc gatgcaccac ccccatctgg tgacagtgta gcacagcctc caggatctgc
     3661 tggatacagt ggctggcatc agcctcactg taatactccc gggccacaat gtcttcgaac
     3721 agctccccac cagtgaccag atcgaagata aggtagtggt gcccctcctc ggagatgctg
     3781 tcatggagtc ggacgatatt ggggtgcttc aacaagcggc agatgcgggc ctcgcgttcc
     3841 aacttctggt gatctctggc tgagagcttc ttggtgttga taatcttggc agcatactcc
     3901 tggccagcca gcaccttcac acacctgcgc accacggaga aggctccctt tcccagttcc
     3961 tcgaagagct ggtactcttc cgtgaatcgg gtgcaggtga tggtagccat agctcgagat
     4021 ctgagaccag acttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac
     4081 tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagctt ggactggtag
     4141 ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg
     4201 tagtggtcgg cgagctgcac gccgccgtcc tcgatgttgt ggcggatctt gaagttggcc
     4261 ttgatgccgt tcttctgctt gtcggcggtg atatagacgt tgtggctgtt ccagttgtac
     4321 tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg
     4381 cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg
     4441 tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag
     4501 tcgtgctgct tcatgtggtc ggggtagcgg gcgaagcact gcaggccgta gcccagggtg
     4561 gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatcag cttcagggtc
     4621 agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt
     4681 acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg
     4741 ctcaccatgg tggcgacctg tagcgctccg gacttgtaca gctcgtccat gccgagagtg
     4801 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     4861 ttgctcagct tggactggta gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     4921 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg
     4981 tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
     5041 ttgtggctgt tccagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     5101 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     5161 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     5221 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     5281 tgcaggccgt agcccagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     5341 gtgcagatca gcttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     5401 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     5461 gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtagcgctag cgaattcttt
     5521 gccaaaatga tgagacagca caataaccag cacgttgccc aggagctgta ggaaaaagaa
     5581 gaaggcatga acatggttag cagaggctct agctagagcc gccggtcaca cgccagaagc
     5641 cgaaccccgc cctgccccgt cccccccgaa ggcagccgtc cccccgcgga cagccccgag
     5701 gctggagagg gagaagggga cggcggcgcg gcgacgcacg aaggccctcc ccgcccattt
     5761 ccttcctgcc ggcgccgcac cgcttcgccc cgcgcccgct agagggggtg cggcggcgcc
     5821 tcccagattt cggctccgca cagatttggg acaaaggaag tccctgcgcc ctctcgcacg
     5881 attaccataa aaggcaatgg ctgcggctcg ccgcgcctcg acagccgccg gcgctccggg
     5941 ggccgccgcg cccctccccc gagccctccc cggcccgagg cggccccgcc ccgcccggca
     6001 cccccacctg ccgccacccc ccgcccggca cggcgagccc cgcgccacgc cccgtacgga
     6061 gccccgcacc cgaagccggg ccgtgctcag caactcgggg aggggggtgc agggggggtt
     6121 gcagcccgac cgacgcgccc acaccccctg ctcacccccc cacgcacaca ccccgcacgc
     6181 agcctttgtt cccctcgcag ccccccccgc accgcggggc accgcccccg gccgcgctcc
     6241 cctcgcgcac actgcggagc gcacaaagcc ccgcgccgcg cccgcagcgc tcacagccgc
     6301 cgggcagcgc ggagccgcac gcggcgctcc ccacgcacac acacacgcac gcaccccccg
     6361 agccgctccc cccgcacaaa gggccctccc ggagcccctc aaggctttca cgcagccaca
     6421 gaaaagaaac aagccgtcat taaaccaagc gctaattaca gcccggagga gaagggccgt
     6481 cccgcccgct cacctgtggg agtaacgcgg tcagtcagag ccggggcggg cggcgcgagg
     6541 cggcggcgga gcggggcacg gggcgaaggc agcgcgcagc gactcccgcc cgccgcgcgc
     6601 ttcgcttttt atagggccgc cgccgccgcc gcctcgccat aaaaggaaac tttcggagcg
     6661 cgccgctctg attggctgcc gccgcacctc tccgcctcgc cccgccccgc ccctcgcccc
     6721 gccccgcccc gcctggcgcg cgcccccccc ccccccccgc ccccatcgct gcacaaaata
     6781 attaaaaaat aaataaatac aaaattgggg gtggggaggg gggggagatg gggagagtga
     6841 agcagaacgt ggggctcacc tcgaccatgg taatagcgat gactaatacg tagatgtact
     6901 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     6961 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     7021 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     7081 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     7141 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     7201 accccgtaat tgattactat taataactag tcaataatca atgtcgatcg acccaggtgg
     7261 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     7321 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     7381 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     7441 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     7501 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     7561 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     7621 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     7681 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     7741 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     7801 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     7861 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     7921 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     7981 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     8041 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     8101 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     8161 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     8221 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     8281 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     8341 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     8401 gatcaaagga tcttc
//
