LOCUS       sequence-89557-e        6625 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric (Blue to Green) fluorescent indicator for calcium 
            signaling in the endoplasmic reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6625)
  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 6625)
  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..6625
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      4..459
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(91..110)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     301..322
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        709..1012
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1015..1290
                     /label=chicken beta-actin promoter
     intron          1291..2299
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     2223..2245
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2355..2371
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2356..2372
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      2381..2390
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     join(2387..2431,2514..2525)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(2387..2432,2515..2525)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             2562..2618
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     2831..2852
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3216..3235)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             3801..3830
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3964..3983)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4029..4084
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4083..4102)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4437..4457)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4437..4455)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4476..4492)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4476..4492)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4489..4511)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4500..4516
                     /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(4524..4530,4531..4548,4549..4554))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4569..4590
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4707..4724)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4878..5466)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4958..4977)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5637..6428,6429..6497))
                     /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     6260..6279
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6498..6602)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
       61 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      121 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      181 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      241 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      301 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      361 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      421 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccta
      661 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      721 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      781 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggacta
      841 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      901 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      961 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tgggtcgagg
     1021 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt
     1081 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggcgcgcg
     1141 ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca
     1201 gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg
     1261 ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg cgttgccttc gccccgtgcc
     1321 ccgctccgcg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     1381 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
     1441 ctcgtttctt ttctgtggct gcgtgaaagc cttaaagggc tccgggaggg ccctttgtgc
     1501 gggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg
     1561 cccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     1621 gtgtgcgcga ggggagcgcg gccgggggcg gtgccccgcg gtgcgggggg gctgcgaggg
     1681 gaacaaaggc tgcgtgcggg gtgtgtgcgt gggggggtga gcagggggtg tgggcgcggc
     1741 ggtcgggctg taaccccccc ctgcaccccc ctccccgagt tgctgagcac ggcccggctt
     1801 cgggtgcggg gctccgtgcg gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc
     1861 ggcaggtggg ggtgccgggc ggggcggggc cgcctcgggc cggggagggc tcgggggagg
     1921 ggcgcggcgg ccccggagcg ccggcggctg tcgaggcgcg gcgagccgca gccattgcct
     1981 tttatggtaa tcgtgcgaga gggcgcaggg acttcctttg tcccaaatct ggcggagccg
     2041 aaatctggga ggcgccgccg caccccctct agcgggcgcg ggcgaagcgg tgcggcgccg
     2101 gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt ccccttctcc
     2161 atctccagcc tcggggctgc cgcaggggga cggctgcctt cgggggggac ggggcagggc
     2221 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     2281 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt gttgtgctgt ctcatcattt
     2341 tggcaaagga ttcctcgagg tcgacggtat cgataagctt gccaccatgg gatggagctg
     2401 tatcatcctc ttcttggtag caacagctac aggtaagggg ttaacagtag caggcttgag
     2461 gtctggacat atatatgggt gacaatgaca tccactttgc ctttctctcc acaggcgcgc
     2521 actcccaggt ccaactgcag ggatccatgg tcgactcttc acgtcgtaag tggaataaga
     2581 caggtcacgc agtcagagct ataggtcggc tgagctcacc agagaacgtg tatataaagg
     2641 ccgacgagca gaagaacggc atcaaggcgt acttcaagat ccgccacaac atcgagggcg
     2701 gcggcgtgca gctcgcctac cactaccagc agatcacccc cataggcgac ggccccgtgc
     2761 tgctgcccga caaccactac ctgagcgtgc agtccatact ttcgaaagac cccaacgaga
     2821 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     2881 acgagctgta caagggtggc agcggtggca tggtgagcaa gggcgaggag ctgttcaccg
     2941 gggtggtgcc catccaggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
     3001 ccggcgaggg tgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
     3061 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgtcctac ggcgtgcagt
     3121 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
     3181 aaggctacat ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     3241 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     3301 tcaaggagga cggcaacatc ctggggcaca agctggagta cagcacgcgt gaccaactga
     3361 ctgaagagca gatcgcagaa tttaaagagg ctttctccct atttgacaag gacggggatg
     3421 ggacgataac aaccaaggat ctggggacgg tgatgcggtc tctggggcag aaccccacag
     3481 aagcagagct ccaggacatg atcaatgaag tagatgccga cggtgacggc acaatcgact
     3541 tccctgagtt cctgacaatg atggcaccta aaatgcagga cacagacagt gaagaggaaa
     3601 ttcgcgaagc gttccgtgtg tgggataagg atggcaatgg ctacatcggc gcagcagagc
     3661 ttcgccacgt gatgacaaac cttggagaga agttaacaga tgaagaggtt gatgaaatga
     3721 tcagggtagc agacatcgat ggagaaggtc aggtaaacta cgaagacttt gtacaaatga
     3781 tgacagcgaa ggcggccgca gaacaaaaac tcatctcaga agaggatctg aatggggccg
     3841 caagcgagaa ggacgagctg tagactagtt ctagagcggc cgcttcactc ctcaggtgca
     3901 ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg
     3961 agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac
     4021 ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct
     4081 ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt
     4141 ttagagtttg gcaacatatg ccatatgctg gctgccatga acaaaggtgg ctataaagag
     4201 gtcatcagta tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga
     4261 cttgaggtta gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa
     4321 aattttcctt acatgtttta ctagccagat ttttcctcct ctcctgacta ctcccagtca
     4381 tagctgtccc tcttctctta tgaagatccc tcgacctgga gctccagctt ttgttccctt
     4441 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     4501 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     4561 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     4621 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     4681 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     4741 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     4801 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4861 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4921 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     4981 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5041 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5101 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5161 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5221 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5281 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5341 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5401 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5461 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5521 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     5581 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     5641 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     5701 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     5761 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     5821 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     5881 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     5941 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     6001 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     6061 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     6121 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     6181 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     6241 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     6301 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     6361 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     6421 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     6481 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     6541 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     6601 cgcacatttc cccgaaaagt gccac
//
