LOCUS       sequence-194349-        5474 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Ratiometric (Blue to Green) fluorescent indicator for calcium 
            signaling in the endoplasmic reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5474)
  AUTHORS   Suzuki J, Kanemaru K, Ishii K, Ohkura M, Okubo Y, Iino M
  TITLE     Imaging intraorganellar Ca(2+) at subcellular resolution using 
            CEPIA.
  JOURNAL   Nat Commun. 2014 Jun 13;5:4153. doi: 10.1038/ncomms5153.
  PUBMED    24923787
REFERENCE   2  (bases 1 to 5474)
  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: "2014-06-13"; volume: "5:4153. doi"; pages: " 
            10.1038/ncomms5153"
COMMENT     Sequence Label: pCIS GEM-CEPIA1er
FEATURES             Location/Qualifiers
     source          1..5474
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     36..58
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     120..139
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     168..184
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     169..185
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      194..203
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     join(200..244,327..338)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(200..245,328..338)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             375..431
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     644..665
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1029..1048)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1614..1643
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1777..1796)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1842..1897
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1896..1915)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     promoter        complement(2252..2270)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2291..2307)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2291..2307)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2304..2326)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2315..2331
                     /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(2339..2345,2346..2363,2364..2369))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2384..2405
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2522..2539)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2693..3281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2773..2792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3452..4243,4244..4312))
                     /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     4075..4094
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4313..4417)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4444..4899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4531..4550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4741..4762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5025..5047
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5039..5056
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5040..5056
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5063..5082
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5063..5081
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        5146..5449
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
       61 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
      121 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa ggattcctcg aggtcgacgg
      181 tatcgataag cttgccacca tgggatggag ctgtatcatc ctcttcttgg tagcaacagc
      241 tacaggtaag gggttaacag tagcaggctt gaggtctgga catatatatg ggtgacaatg
      301 acatccactt tgcctttctc tccacaggcg cgcactccca ggtccaactg cagggatcca
      361 tggtcgactc ttcacgtcgt aagtggaata agacaggtca cgcagtcaga gctataggtc
      421 ggctgagctc accagagaac gtgtatataa aggccgacga gcagaagaac ggcatcaagg
      481 cgtacttcaa gatccgccac aacatcgagg gcggcggcgt gcagctcgcc taccactacc
      541 agcagatcac ccccataggc gacggccccg tgctgctgcc cgacaaccac tacctgagcg
      601 tgcagtccat actttcgaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
      661 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt ggcagcggtg
      721 gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatccag gtcgagctgg
      781 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct
      841 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      901 ccctcgtgac caccctgtcc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      961 agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct
     1021 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1081 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1141 acaagctgga gtacagcacg cgtgaccaac tgactgaaga gcagatcgca gaatttaaag
     1201 aggctttctc cctatttgac aaggacgggg atgggacgat aacaaccaag gatctgggga
     1261 cggtgatgcg gtctctgggg cagaacccca cagaagcaga gctccaggac atgatcaatg
     1321 aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcac
     1381 ctaaaatgca ggacacagac agtgaagagg aaattcgcga agcgttccgt gtgtgggata
     1441 aggatggcaa tggctacatc ggcgcagcag agcttcgcca cgtgatgaca aaccttggag
     1501 agaagttaac agatgaagag gttgatgaaa tgatcagggt agcagacatc gatggagaag
     1561 gtcaggtaaa ctacgaagac tttgtacaaa tgatgacagc gaaggcggcc gcagaacaaa
     1621 aactcatctc agaagaggat ctgaatgggg ccgcaagcga gaaggacgag ctgtagacta
     1681 gttctagagc ggccgcttca ctcctcaggt gcaggctgcc tatcagaagg tggtggctgg
     1741 tgtggccaat gccctggctc acaaatacca ctgagatctt tttccctctg ccaaaaatta
     1801 tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga aatttatttt
     1861 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     1921 aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat atgcccatat
     1981 gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga aacagccccc
     2041 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     2101 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     2161 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
     2221 atccctcgac ctggagctcc agcttttgtt ccctttagtg agggttaatt tcgagcttgg
     2281 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2341 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2401 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     2461 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2521 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2581 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2641 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2701 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2761 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2821 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2881 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2941 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3001 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3061 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3121 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3181 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     3241 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     3301 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     3361 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     3421 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3481 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3541 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3601 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3661 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3721 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3781 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3841 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3901 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3961 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4021 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4081 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4141 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4201 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     4261 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     4321 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     4381 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     4441 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4501 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4561 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4621 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4681 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4741 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4801 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4861 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
     4921 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     4981 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     5041 taaaacgacg gccagtgaat tgtaatacga ctcactatag ggcgaattgg gtacctagta
     5101 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     5161 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     5221 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     5281 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     5341 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     5401 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     5461 ccccacgttc tgct
//
