LOCUS       sequence-325984-        6500 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tdTomato and photoactivatable CaMKII(T286A: 
            Autophosphorylation deficient) in mammalian cells to induce synaptic
            plasticity..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6500)
  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 6500)
  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-tdTomato-P2A-paCaMKII(T286A)
FEATURES             Location/Qualifiers
     source          1..6500
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        112..415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        416..619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     570..590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      658..667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             664..2091
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGRYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             2101..2157
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    4067..4188
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4104..4123)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4158..4177
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      4195..4650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4282..4301)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4492..4513
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4706..4810
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4811..4879,4880..5671)
                     /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(5029..5048)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5842..6430
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6331..6350
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcccctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
       61 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      121 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      181 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      241 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      301 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      361 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      421 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      481 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      541 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      601 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggtcgcc
      661 accatggtga gcaagggcga ggaggtcatc aaagagttca tgcgcttcaa ggtgcgcatg
      721 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
      781 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
      841 gacatcctgt ccccccagtt catgtacggc tccaaggcgt acgtgaagca ccccgccgac
      901 atccccgatt acaagaagct gtccttcccc gagggcttca agtgggagcg cgtgatgaac
      961 ttcgaggacg gcggtctggt gaccgtgacc caggactcct ccctgcagga cggcacgctg
     1021 atctacaagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     1081 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     1141 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1201 accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt ggacaccaag
     1261 ctggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga gcgctccgag
     1321 ggccgccacc acctgttcct ggggcatggc accggcagca ccggcagcgg cagctccggc
     1381 accgcctcct ccgaggacaa caacatggcc gtcatcaaag agttcatgcg cttcaaggtg
     1441 cgcatggagg gctccatgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
     1501 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
     1561 gcctgggaca tcctgtcccc ccagttcatg tacggctcca aggcgtacgt gaagcacccc
     1621 gccgacatcc ccgattacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
     1681 atgaacttcg aggacggcgg tctggtgacc gtgacccagg actcctccct gcaggacggc
     1741 acgctgatct acaaggtgaa gatgcgcggc accaacttcc cccccgacgg ccccgtaatg
     1801 cagaagaaga ccatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
     1861 ctgaagggcg agatccacca ggccctgaag ctgaaggacg gcggccgcta cctggtggag
     1921 ttcaagacca tctacatggc caagaagccc gtgcaactgc ccggctacta ctacgtggac
     1981 accaagctgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgagcgc
     2041 tccgagggcc gccaccacct gttcctgtac ggcatggacg agctgtacaa gggaagcgga
     2101 gccactaact tctccctgtt gaaacaagca ggggatgtcg aagagaatcc cgggccagga
     2161 tctcgagcta tggctaccat cacctgcacc cgattcacgg aagagtacca gctcttcgag
     2221 gaactgggaa agggagcctt ctccgtggtg cgcaggtgtg tgaaggtgct ggctggccag
     2281 gagtatgctg ccaagattat caacaccaag aagctctcag ccagagatca ccagaagttg
     2341 gaacgcgagg cccgcatctg ccgcttgttg aagcacccca atatcgtccg actccatgac
     2401 agcatctccg aggaggggca ccactacctt atcttcgatc tggtcactgg tggggagctg
     2461 ttcgaagaca ttgtggcccg ggagtattac agtgaggctg atgccagcca ctgtatccag
     2521 cagatcctgg aggctgtgct acactgtcac cagatggggg tggtgcatcg cgacctgaag
     2581 cctgagaatc tgttgctggc ttcgaagctc aagggtgctg cggtgaagct ggcagacttt
     2641 ggcctggcca tagaggttga gggagagcag caggcatggt ttgggttcgc agggacacct
     2701 ggatacctct ccccagaagt gctgcggaag gacccatacg ggaagcctgt ggacctgtgg
     2761 gcctgtggcg tcatcctgta tatcttgctg gttgggtatc ccccattctg ggatgaggac
     2821 cagcaccgcc tgtaccagca gatcaaagct ggtgcctacg atttcccatc accagaatgg
     2881 gacaccgtca ccccggaagc caaggatctg atcaataaga tgctgaccat caacccgtcc
     2941 aaacgcatca cggccgctga ggctctcaag cacccctgga tctcgcaccg ctccaagctt
     3001 ctggaacgca ttgaaaaaaa ctttgtgatt accgatccgc gcctgccgga taacccgatt
     3061 atttttgcga gcgatagctt tctgcagctg accgaatata gccgcgaaga aattctgggc
     3121 cgcaactgcc gctttctgca gggcccggaa accgatcgcg cgaccgtgcg caaaattcgc
     3181 gatgcgattg ataaccagac cgaagtgacc gtgcagctga ttaactatac caaaagcggc
     3241 aaaaaatttt ggaacctgtt tcatctgcag ccgatgcgcg atcagaaagg cgatgtgcag
     3301 tattttattg gcgtgcagct ggatggcacc gaacatgtgc gcgatgcggc ggaacgcgaa
     3361 ggcgtgatgc tgattaaaaa aaccgcggaa aacattgatg aagcggcgaa agagctcgaa
     3421 ttcgccaagt gcatgcacag acaggaggcc gtggactgcc tgaagaagtt caatgccagg
     3481 aggaaactga agggagccat cctcaccact atgctggcca ccaggaactt ctccgtgcgc
     3541 aaacaggaaa ttatcaaagt gacagagcag ctgatcgaag ccataagcaa tggagacttt
     3601 gaatcctaca cgaagatgtg cgaccctgga atgacagcct ttgaaccgga ggccctgggg
     3661 aacctggtcg agggcctgga ccttcatcga ttctattttg aaaacctgtg gtcccggaac
     3721 agcaagcccg tgcacaccac catcctgaac cctcacgtcc acctgatggg tgacgagtca
     3781 gcctgcatca cctacatccg caccactcag tacctggatg cgggtggcat cccccgcacg
     3841 gcccagtcag aggagacccg tgtctggcac cgcagggatg gaaaatggca gatcgtccac
     3901 ttccacagat caggggcgcc ctccgtcctg ccccatggat cctaagcggc cgcgacacta
     3961 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     4021 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     4081 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4141 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtacgcgc
     4201 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4261 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4321 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4381 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     4441 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4501 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     4561 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4621 attttaacaa aatattaacg cttacaattt cctctagata ataatggttt cttagacgtc
     4681 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4741 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4801 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     4861 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     4921 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4981 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5041 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5101 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5161 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5221 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5281 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     5341 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     5401 tactctagct tcccggcaac aactgataga ctggatggag gcggataaag ttgcaggacc
     5461 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     5521 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     5581 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     5641 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     5701 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5761 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5821 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5881 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5941 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     6001 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6061 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6121 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6181 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6241 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6301 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     6361 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     6421 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6481 tgctcacatg gctcgacaga
//
