LOCUS       sequence-238497-        7158 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  luminescent reporter for calcium signaling.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7158)
  AUTHORS   Suzuki K, Kimura T, Shinoda H, Bai G, Daniels MJ, Arai Y, Nakano M, 
            Nagai T
  TITLE     Five colour variants of bright luminescent protein for real-time 
            multicolour bioimaging.
  JOURNAL   Nat Commun. 2016 Dec 14;7:13718. doi: 10.1038/ncomms13718.
  PUBMED    27966527
REFERENCE   2  (bases 1 to 7158)
  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: "2016-12-14"; volume: "7:13718. doi"; pages: " 
            10.1038/ncomms13718"
COMMENT     Sequence Label: GeNL(Ca2+)_60/pcDNA3
FEATURES             Location/Qualifiers
     source          1..7158
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        149..528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        529..732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     683..703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     777..796
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        777..795
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2163..2240
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     promoter        complement(2625..2643)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2626..2643)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2663..2680)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2669..2893
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2939..3367
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3026..3045)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3236..3257
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3376..3396)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3381..3710
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3561..3696
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3623..3642
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3777..4571
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3831..3850)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4441..4460
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4745..4866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4782..4801)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4836..4855
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4915..4931)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4915..4931)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4928..4950)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4939..4955
                     /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(4963..4969,4970..4987,4988..4993))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5008..5029
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5146..5163)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5317..5905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5397..5416)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6076..6867,6868..6936))
                     /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     6699..6718
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6937..7041)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7116..7135)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
       61 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
      121 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
      181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      721 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
      781 acgactcact atagggagac ccaagcttgg taccgagctc ggatccgatg gtgtccaagg
      841 gcgaagagga caacatggcc agcctgcctg ccacccacga gctgcacatc ttcggcagca
      901 tcaacggcgt ggacttcgac atggtgggac agggcaccgg caaccccaac gacggctacg
      961 aggaactgaa cctgaagtcc acaaagggcg acctgcagtt cagcccctgg attctggtgc
     1021 cccacatcgg ctacggcttc caccagtacc tgccctaccc cgacggcatg agccctttcc
     1081 aggccgctat ggtggatggc agcggctacc aggtgcaccg gaccatgcag tttgaggacg
     1141 gcgccagcct gaccgtgaac taccggtaca catacgaggg cagccacatc aagggcgagg
     1201 cccaagtgaa gggcacaggc tttccagccg acggccccgt gatgaccaat agcctgacag
     1261 ccgccgactg gtgcagaagc aagaaaacct accccaatga caagaccatc atcagcacct
     1321 tcaagtggtc ctacaccacc ggcaatggca agcggtacag aagcaccgcc cggaccacct
     1381 acaccttcgc caaacctatg gccgccaact acctgaagaa ccagcctatg tacgtgttcc
     1441 gcaagaccga gctgaagcac tccaagacag aactgaactt caaagagtgg cagaaagcct
     1501 tcaccgggtt tgaagatttc gttggggact ggcgacagac agccggctac aacctggacc
     1561 aagtccttga acagggaggt gtgtccagtt tgtttcagaa tctcggggtg tccgtaactc
     1621 cgatccaaag gattgtcctg agcggtgaaa atgggctgaa gatcgacatc catgtcatca
     1681 tcccgtatga aggtccatgg atgcatgacc aactgacaga agagcagatt gcagagttca
     1741 aagaagcctt ctcattattc gacaaggatg gggacggcac catcaccaca agggaacttg
     1801 gcaccgttat gaggtcgctt ggacaaaacc caacggaagc cgaattgcag gatatgatca
     1861 atgaagtcga tgctgatggc aatggaacga tttactttcc tgaatttctt actatgatgg
     1921 ctagaaaaat gaaggacaca gacagcgaag aggaaatccg agaagcattc cgtgtttttg
     1981 acaaggatgg gaacggctac atcagcgctg ctgaattacg tcacgtcatg acaaacctcg
     2041 gggtgaagtt aacagatgaa gaagttgatg aaatgataag ggaagcagat atcgatggtg
     2101 atggccaagt aaactatgaa gagtttgaac aaatgatgac agcaaagggg ggaggtggct
     2161 ccaagaggcg ctggaagaaa aacttcattg ccgtcagcgc tgccaaccgg ttcaagaaga
     2221 tctccagctc cggggcactg gagctcctga gcggcgacca aatgggccag atcgaaaaaa
     2281 tttttaaggt ggtgtaccct gtggatgatc atcactttaa ggtgatcctg cactatggca
     2341 cactggtaat cgacggggtt acgccgaaca tgatcgacta tttcggacgg ccgtatgaag
     2401 gcatcgccgt gttcgacggc aaaaagatca ctgtaacagg gaccctgtgg aacggcaaca
     2461 aaattatcga cgagcgcctg atcaaccccg acggctccct gctgttccga gtaaccatca
     2521 acggagtgac cggctggcgg ctgtgcgaac gcattctggc gtaagaattc tgcagatatc
     2581 catcacactg gcggccgctc gagcatgcat ctagagggcc ctattctata gtgtcaccta
     2641 aatgctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2701 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2761 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2821 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2881 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2941 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3001 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3061 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3121 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3181 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3241 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3301 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3361 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3421 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3481 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3541 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3601 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3661 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3721 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     3781 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     3841 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     3901 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     3961 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4021 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4081 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4141 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4201 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4261 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4321 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4381 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4441 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4501 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4561 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     4621 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4681 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4741 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4801 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4861 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4921 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4981 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     5041 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     5101 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     5161 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     5221 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     5281 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     5341 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5401 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5461 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     5521 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5581 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5641 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5701 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     5761 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5821 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5881 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5941 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     6001 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     6061 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     6121 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     6181 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     6241 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     6301 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6361 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6421 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6481 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6541 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6601 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6661 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6721 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6781 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6841 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6901 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6961 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     7021 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     7081 gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     7141 ttaagccagt atctgctc
//
