LOCUS       sequence-250801-        5942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for calmodulin 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5942)
  AUTHORS   Chang JY, Nakahata Y, Hayano Y, Yasuda R
  TITLE     Mechanisms of Ca(2+)/calmodulin-dependent kinase II activation in 
            single dendritic spines.
  JOURNAL   Nat Commun. 2019 Jun 25;10(1):2784. doi: 10.1038/s41467-019-10694-z.
  PUBMED    31239443
REFERENCE   2  (bases 1 to 5942)
  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: "2019-06-25"; volume: "10(1):2784. doi"; pages: " 
            10.1038/s41467-019-10694-z"
COMMENT     Sequence Label: pCAG-mCherry-CaM
FEATURES             Location/Qualifiers
     source          1..5942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          261..1281
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1201..1223
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1289..1308
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1342..1351
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1348..2055
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1493..1511)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1761..1780
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1955..1978
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     CDS             2080..2529
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPT
                     EAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(2556..2575)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2621..2676
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3037..3053)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3037..3053)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3050..3072)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3061..3077
                     /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(3085..3091,3092..3109,3110..3115))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3130..3151
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3210..3405
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3256..3391
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3411..3545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3448..3467)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3502..3521
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3613..3630)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3784..4372)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3864..3883)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4543..5334,5335..5403))
                     /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     5166..5185
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5404..5508)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5543..5922
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5924..5942,1..260)
                     /label=chicken beta-actin promoter
ORIGIN
        1 ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt tatttatttt
       61 ttaattattt tgtgcagcga tgggggcggg gggggggggg ggggcgcgcg ccgggggggg
      121 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
      181 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
      241 agcgaagcgc gcggcgggcg ggagtcgctg cgcgctgcct tcgccccgtg ccccgctccg
      301 ccgccgcctc gcgccgcccg ccccggctct gactgaccgc gttactccca caggtgagcg
      361 ggcgggacgg cccttctcct ccgggctgta attagcgctt ggtttaatga cggcttgttt
      421 cttttctgtg gctgcgtgaa agccttgagg ggctccggga gggccctttg tgcgggggga
      481 gcggctcggg gggtgcgtgc gtgtgtgtgt gcgtggggag cgccgcgtgc ggctccgcgc
      541 tgcccggcgg ctgtgagcgc tgcgggcgcg gcgcggggct ttgtgcgctc cgcagtgtgc
      601 gcgaggggag cgcggccggg ggcggtgccc cgcggtgcgg ggggggctgc gaggggaaca
      661 aaggctgcgt gcggggtgtg tgcgtggggg ggtgagcagg gggtgtgggc gcgtcggtcg
      721 ggctgcaacc ccccctgcac ccccctcccc gagttgctga gcacggcccg gcttcgggtg
      781 cggggctccg tacggggcgt ggcgcggggc tcgccgtgcc gggcgggggg tggcggcagg
      841 tgggggtgcc gggcggggcg gggccgcctc gggccgggga gggctcgggg gaggggcgcg
      901 gcggcccccg gagcgccggc ggctgtcgag gcgcggcgag ccgcagccat tgccttttat
      961 ggtaatcgtg cgagagggcg cagggacttc ctttgtccca aatctgtgcg gagccgaaat
     1021 ctgggaggcg ccgccgcacc ccctctagcg ggcgcggggc gaagcggtgc ggcgccggca
     1081 ggaaggaaat gggcggggag ggccttcgtg cgtcgccgcg ccgccgtccc cttctccctc
     1141 tccagcctcg gggctgtccg cggggggacg gctgccttcg ggggggacgg ggcagggcgg
     1201 ggttcggctt ctggcgtgtg accggcggct ctagctagag cctctgctaa ccatgttcat
     1261 gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat
     1321 tttggcaaag aattcgctag cgccaccatg gtgagcaagg gcgaggagga caacatggcc
     1381 atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag
     1441 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1501 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg
     1561 tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc
     1621 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc
     1681 gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc
     1741 accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1801 tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag
     1861 ctgaaggacg gcggccacta cgacgccgag gtcaagacca cctacaaggc caagaagccc
     1921 gtgcagctgc ccggcgccta caacgtcaac atcaagctgg acatcacctc ccacaacgag
     1981 gactacacca tcgtggaaca gtacgagcgc gccgagggcc gccactccac cggcggcatg
     2041 gacgagctgt acaagtccgg actcagatct cgagctcaaa tggctgatca gctgactgaa
     2101 gagcagattg ctgaattcaa ggaagctttc tccctattcg ataaagatgg tgacggcacc
     2161 atcacaacca aggaactggg gaccgtcatg cggtcactgg gtcagaaccc aacagaagcc
     2221 gagctgcagg atatgatcaa cgaagtggat gctgatggca atggcaccat tgacttccca
     2281 gagttcttga ctatgatggc tagaaaaatg aaagacacag atagcgaaga agagatccgc
     2341 gaggccttcc gagtgtttga caaggatggg aatggttaca tcagtgcggc agaactgcgc
     2401 cacgtcatga caaacttagg agaaaagcta acagatgaag aagtagatga aatgatcaga
     2461 gaagcagata ttgatggcga cggacaagtc aactatgaag aattcgtaca gatgatgact
     2521 gcaaaatgag cggccgcgac tctagagtcg acggatcttt ttccctctgc caaaaattat
     2581 ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa atttattttc
     2641 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     2701 aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata tgcccatatg
     2761 ctggctgcca tgaacaaagg ttggctataa agaggtcatc agtatatgaa acagccccct
     2821 gctgtccatt ccttattcca tagaaaagcc ttgacttgag gttagatttt ttttatattt
     2881 tgttttgtgt tatttttttc tttaacatcc ctaaaatttt ccttacatgt tttactagcc
     2941 agatttttcc tcctctcctg actactccca gtcatagctg tccctcttct cttatggaga
     3001 tccctcgacc tgcagcccaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3061 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3121 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3181 gtcgggaaac ctgtcgtgcc agcggatccg catctcaatt agtcagcaac catagtcccg
     3241 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     3301 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     3361 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     3421 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3481 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3541 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3601 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3661 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3721 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3781 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3841 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3901 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3961 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4021 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4081 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4141 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4201 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4261 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4321 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4381 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4441 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4501 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4561 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4621 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4681 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4741 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4801 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4861 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4921 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4981 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     5041 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     5101 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     5161 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     5221 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     5281 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     5341 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5401 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5461 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5521 aaaagtgcca cctgggtcga tcgacattga ttattgacta gttattaata gtaatcaatt
     5581 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
     5641 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
     5701 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
     5761 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
     5821 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
     5881 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
     5941 tt
//
