LOCUS       sequence-325974-        5732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses photoactivatable CaMKII(K42M: Kinase dead) in mammalian 
            cells as a negative control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5732)
  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 5732)
  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)
FEATURES             Location/Qualifiers
     source          1..5732
                     /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             join(664..666,667..669,670..1380)
                     /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(709..730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(970..989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1317..1338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3299..3420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3336..3355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3390..3409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      3427..3882
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3514..3533)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3724..3745
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3938..4042
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4043..4111,4112..4903)
                     /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(4261..4280)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5074..5662
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5563..5582
                     /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 ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      721 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      781 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      841 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      901 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      961 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1021 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1081 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1141 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1261 cactacctga gcacccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
     1321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1381 tccggactca gatctcgagc tatggctacc atcacctgca cccgattcac ggaagagtac
     1441 cagctcttcg aggaactggg aaagggagcc ttctccgtgg tgcgcaggtg tgtgaaggtg
     1501 ctggctggcc aggagtatgc tgccatgatt atcaacacca agaagctctc agccagagat
     1561 caccagaagt tggaacgcga ggcccgcatc tgccgcttgt tgaagcaccc caatatcgtc
     1621 cgactccatg acagcatctc cgaggagggg caccactacc ttatcttcga tctggtcact
     1681 ggtggggagc tgttcgaaga cattgtggcc cgggagtatt acagtgaggc tgatgccagc
     1741 cactgtatcc agcagatcct ggaggctgtg ctacactgtc accagatggg ggtggtgcat
     1801 cgcgacctga agcctgagaa tctgttgctg gcttcgaagc tcaagggtgc tgcggtgaag
     1861 ctggcagact ttggcctggc catagaggtt gagggagagc agcaggcatg gtttgggttc
     1921 gcagggacac ctggatacct ctccccagaa gtgctgcgga aggacccata cgggaagcct
     1981 gtggacctgt gggcctgtgg cgtcatcctg tatatcttgc tggttgggta tcccccattc
     2041 tgggatgagg accagcaccg cctgtaccag cagatcaaag ctggtgccta cgatttccca
     2101 tcaccagaat gggacaccgt caccccggaa gccaaggatc tgatcaataa gatgctgacc
     2161 atcaacccgt ccaaacgcat cacggccgct gaggctctca agcacccctg gatctcgcac
     2221 cgctccaagc ttctggaacg cattgaaaaa aactttgtga ttaccgatcc gcgcctgccg
     2281 gataacccga ttatttttgc gagcgatagc tttctgcagc tgaccgaata tagccgcgaa
     2341 gaaattctgg gccgcaactg ccgctttctg cagggcccgg aaaccgatcg cgcgaccgtg
     2401 cgcaaaattc gcgatgcgat tgataaccag accgaagtga ccgtgcagct gattaactat
     2461 accaaaagcg gcaaaaaatt ttggaacctg tttcatctgc agccgatgcg cgatcagaaa
     2521 ggcgatgtgc agtattttat tggcgtgcag ctggatggca ccgaacatgt gcgcgatgcg
     2581 gcggaacgcg aaggcgtgat gctgattaaa aaaaccgcgg aaaacattga tgaagcggcg
     2641 aaagagctcg aattcgccaa gtgcatgcac agacaggaga ccgtggactg cctgaagaag
     2701 ttcaatgcca ggaggaaact gaagggagcc atcctcacca ctatgctggc caccaggaac
     2761 ttctccgtgc gcaaacagga aattatcaaa gtgacagagc agctgatcga agccataagc
     2821 aatggagact ttgaatccta cacgaagatg tgcgaccctg gaatgacagc ctttgaaccg
     2881 gaggccctgg ggaacctggt cgagggcctg gaccttcatc gattctattt tgaaaacctg
     2941 tggtcccgga acagcaagcc cgtgcacacc accatcctga accctcacgt ccacctgatg
     3001 ggtgacgagt cagcctgcat cacctacatc cgcaccactc agtacctgga tgcgggtggc
     3061 atcccccgca cggcccagtc agaggagacc cgtgtctggc accgcaggga tggaaaatgg
     3121 cagatcgtcc acttccacag atcaggggcg ccctccgtcc tgccccatgg atcctaagcg
     3181 gccgcgacac tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3241 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3301 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3361 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3421 acgcgtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     3481 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     3541 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     3601 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     3661 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     3721 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     3781 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     3841 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttcctctaga taataatggt
     3901 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3961 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4021 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4081 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4141 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4201 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4261 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4321 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4381 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4441 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4501 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4561 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     4621 tggcgaacta cttactctag cttcccggca acaactgata gactggatgg aggcggataa
     4681 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     4741 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     4801 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     4861 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     4921 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4981 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5041 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5101 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5161 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5221 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5281 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5341 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5401 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5461 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5521 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5581 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5641 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5701 ttgctggcct tttgctcaca tggctcgaca ga
//
