LOCUS       sequence-238498-        7155 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 7155)
  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 7155)
  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+)_250/pcDNA3
FEATURES             Location/Qualifiers
     source          1..7155
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        121..500
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        501..704
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     655..675
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     749..768
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        749..767
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2132..2209
                     /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(2594..2612)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2595..2612)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2632..2649)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2638..2862
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2908..3336
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2995..3014)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3205..3226
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3345..3365)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3350..3679
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3530..3665
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3592..3611
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3746..4540
                     /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(3800..3819)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4410..4429
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4714..4835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4751..4770)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4805..4824
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4884..4900)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4884..4900)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4897..4919)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4908..4924
                     /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(4932..4938,4939..4956,4957..4962))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4977..4998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5115..5132)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5286..5874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5366..5385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6045..6836,6837..6905))
                     /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     6668..6687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6906..7010)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7085..7104)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
       61 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      121 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      181 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      241 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      301 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      361 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      421 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      481 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      541 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      601 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      661 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      721 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      781 ggtaccgagc tcggatccga tggtgtccaa gggcgaagag gacaacatgg ccagcctgcc
      841 tgccacccac gagctgcaca tcttcggcag catcaacggc gtggacttcg acatggtggg
      901 acagggcacc ggcaacccca acgacggcta cgaggaactg aacctgaagt ccacaaaggg
      961 cgacctgcag ttcagcccct ggattctggt gccccacatc ggctacggct tccaccagta
     1021 cctgccctac cccgacggca tgagcccttt ccaggccgct atggtggatg gcagcggcta
     1081 ccaggtgcac cggaccatgc agtttgagga cggcgccagc ctgaccgtga actaccggta
     1141 cacatacgag ggcagccaca tcaagggcga ggcccaagtg aagggcacag gctttccagc
     1201 cgacggcccc gtgatgacca atagcctgac agccgccgac tggtgcagaa gcaagaaaac
     1261 ctaccccaat gacaagacca tcatcagcac cttcaagtgg tcctacacca ccggcaatgg
     1321 caagcggtac agaagcaccg cccggaccac ctacaccttc gccaaaccta tggccgccaa
     1381 ctacctgaag aaccagccta tgtacgtgtt ccgcaagacc gagctgaagc actccaagac
     1441 agaactgaac ttcaaagagt ggcagaaagc cttcaccggg tttgaagatt tcgttgggga
     1501 ctggcgacag acagccggct acaacctgga ccaagtcctt gaacagggag gtgtgtccag
     1561 tttgtttcag aatctcgggg tgtccgtaac tccgatccaa aggattgtcc tgagcggtga
     1621 aaatgggctg aagatcgaca tccatgtcat catcccgtat gaaggtccat ggatgcatga
     1681 ccaactgaca gaagagcaga ttgcagagtt caaagaagcc ttctcattat tcgacaagga
     1741 tggggacggc accatcacca caagggaact tggcaccgtt atgaggtcgc ttggacaaaa
     1801 cccaacggaa gccgaattgc aggatatgat caatgaagtc gatgctgatg gcaatggaac
     1861 gatttacttt cctgaatttc ttactatgat ggctagaaaa atgaaggaca cagacagcga
     1921 agaggaaatc cgagaagcat tccgtgtttt tgacaaggat gggaacggct acatcagcgc
     1981 tgctcagtta cgtcacgtca tgacaaacct cggggtgaag ttaacagatg aagaagttga
     2041 tgaaatgata agggaagcag atatcgatgg tgatggccaa gtaaactatg aagagtttga
     2101 acaaatgatg acagcaaagg gaggtggctc caagaggcgc tggaagaaaa acttcattgc
     2161 cgtcagcgct gccaaccggt tcaagaagat ctccagctcc ggggcactgg agctcctgag
     2221 cggcgaccaa atgggccaga tcgaaaaaat ttttaaggtg gtgtaccctg tggatgatca
     2281 tcactttaag gtgatcctgc actatggcac actggtaatc gacggggtta cgccgaacat
     2341 gatcgactat ttcggacggc cgtatgaagg catcgccgtg ttcgacggca aaaagatcac
     2401 tgtaacaggg accctgtgga acggcaacaa aattatcgac gagcgcctga tcaaccccga
     2461 cggctccctg ctgttccgag taaccatcaa cggagtgacc ggctggcggc tgtgcgaacg
     2521 cattctggcg taagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc
     2581 tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg
     2641 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     2701 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     2761 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     2821 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
     2881 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
     2941 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3001 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3061 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3121 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3181 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3241 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3301 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     3361 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3421 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     3481 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     3541 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3601 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     3661 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     3721 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3781 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     3841 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     3901 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     3961 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4021 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4081 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4141 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4201 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4261 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4321 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4381 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4441 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4501 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4561 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4621 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4681 ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt
     4741 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4801 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct
     4861 agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     4921 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     4981 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     5041 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     5101 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5161 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5221 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     5281 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     5341 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     5401 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     5461 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     5521 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     5581 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     5641 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     5701 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     5761 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     5821 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     5881 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     5941 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6001 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6061 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6121 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6181 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6241 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     6301 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     6361 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     6421 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     6481 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     6541 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     6601 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     6661 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     6721 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     6781 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     6841 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     6901 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6961 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7021 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
     7081 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
     7141 tggaggtcgc tgagt
//
