LOCUS       sequence-477254-        5262 bp    DNA     circular SYN 26-JUN-2025
DEFINITION  Calmodulin-1 for CaMK2-alpha sensor assays in mammalian cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5262)
  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 5262)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 26, 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-Calmodulin1 (CaM)
FEATURES             Location/Qualifiers
     source          1..5262
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..667
                     /label=chicken beta-actin promoter
     intron          668..1688
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1608..1630
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1696..1715
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1749..1758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1755..2204
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="MADQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPT
                     EAELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYISAA
                     ELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     complement(2284..2303)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2349..2404
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2765..2781)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2765..2781)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2778..2800)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2789..2805
                     /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(2813..2819,2820..2837,2838..2843))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2858..2879
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2938..3133
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2984..3119
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3139..3273
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3176..3195)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3230..3249
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3341..3358)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3512..4100)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3592..3611)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4271..5062,5063..5131))
                     /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     4894..4913
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5132..5236)
                     /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 gggcgggcca ggcggggcgg ggcggggcga
      541 ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg
      601 aaagtttcct tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg
      661 gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg
      721 ccgcccgccc cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc
      781 ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct
      841 gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcgggggg
      901 tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg
      961 tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc
     1021 ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg
     1081 gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc
     1141 cctgcacccc cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtac
     1201 ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg
     1261 cggggcgggg ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag
     1321 cgccggcggc tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga
     1381 gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg
     1441 ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg
     1501 cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg
     1561 ctgtccgcgg ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg
     1621 gcgtgtgacc ggcggctcta gctagagcct ctgctaacca tgttcatgcc ttcttctttt
     1681 tcctacagct cctgggcaac gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat
     1741 tcgctagcgc caccatggct gatcagctga ctgaagagca gattgctgaa ttcaaggaag
     1801 ctttctccct attcgataaa gatggtgacg gcaccatcac aaccaaggaa ctggggaccg
     1861 tcatgcggtc actgggtcag aacccaacag aagccgagct gcaggatatg atcaacgaag
     1921 tggatgctga tggcaatggc accattgact tcccagagtt cttgactatg atggctagaa
     1981 aaatgaaaga cacagatagc gaagaagaga tccgcgaggc cttccgagtg tttgacaagg
     2041 atgggaatgg ttacatcagt gcggcagaac tgcgccacgt catgacaaac ttaggagaaa
     2101 agctaacaga tgaagaagta gatgaaatga tcagagaagc agatattgat ggcgacggac
     2161 aagtcaacta tgaagaattc gtacagatga tgactgcaaa atgagagctc aagcttcgaa
     2221 ttctgcagtc gacggtaccg cgggcccggg atcctaagcg gccgcgactc tagagtcgac
     2281 ggatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     2341 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     2401 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     2461 ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt ggctataaag
     2521 aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata gaaaagcctt
     2581 gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt taacatccct
     2641 aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac tactcccagt
     2701 catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc ttggcgtaat
     2761 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     2821 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     2881 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag cggatccgca
     2941 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3001 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3061 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3121 aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3181 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3241 aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg
     3301 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     3361 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     3421 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3481 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3541 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3601 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3661 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3721 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3781 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3841 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3901 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3961 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     4021 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     4081 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     4141 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     4201 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     4261 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     4321 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     4381 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     4441 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     4501 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     4561 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     4621 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     4681 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     4741 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     4801 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     4861 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     4921 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4981 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     5041 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     5101 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     5161 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     5221 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tg
//
