LOCUS       sequence-238494-        7152 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 7152)
  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 7152)
  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+)_480/pcDNA3
FEATURES             Location/Qualifiers
     source          1..7152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        143..522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        523..726
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     677..697
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     771..790
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        771..789
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2151..2228
                     /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(2613..2631)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2614..2631)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2651..2668)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2657..2881
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2927..3355
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3014..3033)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3224..3245
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3364..3384)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3369..3698
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3549..3684
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3611..3630
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3765..4559
                     /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(3819..3838)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4429..4448
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4733..4854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4770..4789)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4824..4843
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4903..4919)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4903..4919)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4916..4938)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4927..4943
                     /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(4951..4957,4958..4975,4976..4981))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4996..5017
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5134..5151)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5305..5893)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5385..5404)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6064..6855,6856..6924))
                     /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     6687..6706
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6925..7029)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7104..7123)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac aaggcaaggc
       61 ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct gcttcgcgat
      121 gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata gtaatcaatt
      181 acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat
      241 ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt
      301 cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
      361 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
      421 aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct
      481 acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag
      541 tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt
      601 gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaac
      661 aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc
      721 agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat taatacgact
      781 cactataggg agacccaagc ttggtaccga gctcggatcc gatggtgtcc aagggcgaag
      841 aggacaacat ggccagcctg cctgccaccc acgagctgca catcttcggc agcatcaacg
      901 gcgtggactt cgacatggtg ggacagggca ccggcaaccc caacgacggc tacgaggaac
      961 tgaacctgaa gtccacaaag ggcgacctgc agttcagccc ctggattctg gtgccccaca
     1021 tcggctacgg cttccaccag tacctgccct accccgacgg catgagccct ttccaggccg
     1081 ctatggtgga tggcagcggc taccaggtgc accggaccat gcagtttgag gacggcgcca
     1141 gcctgaccgt gaactaccgg tacacatacg agggcagcca catcaagggc gaggcccaag
     1201 tgaagggcac aggctttcca gccgacggcc ccgtgatgac caatagcctg acagccgccg
     1261 actggtgcag aagcaagaaa acctacccca atgacaagac catcatcagc accttcaagt
     1321 ggtcctacac caccggcaat ggcaagcggt acagaagcac cgcccggacc acctacacct
     1381 tcgccaaacc tatggccgcc aactacctga agaaccagcc tatgtacgtg ttccgcaaga
     1441 ccgagctgaa gcactccaag acagaactga acttcaaaga gtggcagaaa gccttcaccg
     1501 ggtttgaaga tttcgttggg gactggcgac agacagccgg ctacaacctg gaccaagtcc
     1561 ttgaacaggg aggtgtgtcc agtttgtttc agaatctcgg ggtgtccgta actccgatcc
     1621 aaaggattgt cctgagcggt gaaaatgggc tgaagatcga catccatgtc atcatcccgt
     1681 atgaaggtcc atggatgcat gaccaactga cagaagagca gattgcagag ttcaaagaag
     1741 ccttctcatt attcgacaag gatggggacg gcaccatcac cacaagggaa cttggcaccg
     1801 ttatgaggtc gcttggacaa aacccaacgg aagccgaatt gcaggatatg atcaatgaag
     1861 tcgatgctga tggcaatgga acgatttact ttcctgaatt tcttactatg atggctagaa
     1921 aaatgaagga cacagacagc gaagaggaaa tccgagaagc attccgtgtt tttgacaagg
     1981 atgggaacgg ctacatcagc gctgctcagt tacgtcacgt catgacaaac ctcggggtga
     2041 agttaacaga tgaagaagtt gatgaaatga taagggaagc agatatcgat ggtgatggcc
     2101 aagtaaacta tgaagagttt gaacaaatga tgacagcaaa gggtggctcc aagaggcgct
     2161 ggaagaaaaa cttcattgcc gtcagcgctg ccaaccggtt caagaagatc tccagctccg
     2221 gggcactgga gctcctgagc ggcgaccaaa tgggccagat cgaaaaaatt tttaaggtgg
     2281 tgtaccctgt ggatgatcat cactttaagg tgatcctgca ctatggcaca ctggtaatcg
     2341 acggggttac gccgaacatg atcgactatt tcggacggcc gtatgaaggc atcgccgtgt
     2401 tcgacggcaa aaagatcact gtaacaggga ccctgtggaa cggcaacaaa attatcgacg
     2461 agcgcctgat caaccccgac ggctccctgc tgttccgagt aaccatcaac ggagtgaccg
     2521 gctggcggct gtgcgaacgc attctggcgt aagaattctg cagatatcca tcacactggc
     2581 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     2641 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2701 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2761 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2821 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2881 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2941 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3001 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3061 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3121 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3181 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3241 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3301 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3361 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3421 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3481 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3541 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3601 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3661 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3721 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3781 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3841 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3901 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3961 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4021 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4081 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4141 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4201 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4261 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4321 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4381 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4441 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4501 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4561 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4621 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4681 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4741 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4801 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4861 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4921 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4981 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5041 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5101 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5161 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5221 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5281 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5341 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5401 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5461 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5521 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5581 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5641 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5701 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5761 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5821 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5881 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5941 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6001 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6061 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6121 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6181 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6241 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6301 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6361 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6421 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6481 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6541 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6601 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6661 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6721 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6781 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6841 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6901 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6961 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7021 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7081 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7141 ctgctccctg ct
//
