LOCUS       sequence-89554-e        6299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent indicator for calcium imaging in the endoplasmic 
            reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6299)
  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 6299)
  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: pCMV G-CEPIA1er
FEATURES             Location/Qualifiers
     source          1..6299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        12..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      650..659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     join(656..700,783..794)
                     /codon_start=1
                     /product="modified signal sequence from the mouse 
                     immunoglobulin heavy chain variable region"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             join(656..701,784..794)
                     /codon_start=1
                     /product="signal sequence from the mouse immunoglobulin 
                     heavy chain variableregion"
                     /label=IgH signal sequence
                     /translation="MGWSCIILFLVATATGAHS"
     CDS             831..887
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     1100..1121
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1488..1507)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1623..2063
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKDLGTVLRSLGQNPTEA
                     ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVWDKDGNGYISAADL
                     RHVMTNLGEKLTDEEVDEMIREADIDGEGQVNYEEFVQMMTAK"
     CDS             2073..2102
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(2156..2173)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2162..2386
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2432..2860
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2519..2538)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2729..2750
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2869..2889)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2874..3203
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3054..3189
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3116..3135
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3229..4023
                     /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="MVEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3283..3302)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3893..3912
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4197..4318
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4234..4253)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4288..4307
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4351..4368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4522..5110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4602..4621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5281..6072,6073..6141))
                     /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     5904..5923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6142..6246)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtaccgaatt cacattgatt attgagtagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacggtt agtcatcgct attaccatag tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctttctggc
      601 taactagaga acccactgct tactggcacg tggaaattaa tacgacgtgg ccaccatggg
      661 atggagctgt atcatcctct tcttggtagc aacagctaca ggtaaggggt taacagtagc
      721 aggcttgagg tctggacata tatatgggtg acaatgacat ccactttgcc tttctctcca
      781 caggcgcgca ctcccaggtc caactgcagg gatccatggt cgactcttca cgtcgtaagt
      841 ggaataagac aggtcacgca gtcagagcta taggtcggct gagctcactc gagaacgtct
      901 atatcaaggc cgacgagcag aagaacggca tcaaggcgta cttcaagatc cgccacaaca
      961 tcgagggcgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg
     1021 gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccatactt tcgaaagacc
     1081 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1141 tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgagc aagggcgagg
     1201 agctgttcac cggggtggtg cccatccagg tcgagctgga cggcgacgta aacggccaca
     1261 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt
     1321 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     1381 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     1441 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact
     1501 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     1561 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacacgc
     1621 gtgaccaact gactgaagag cagatcgcag aatttaaaga ggctttctcc ctatttgaca
     1681 aggacgggga tgggacgata acaaccaagg atctggggac ggtgctgcgg tctctggggc
     1741 agaaccccac agaagcagag ctccaggaca tgatcaatga agtagatgcc gacggtaatg
     1801 gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa gacacagaca
     1861 gtgaagaaga aattcgcgaa gcgttccgtg tgtgggataa ggatggcaat ggctacatct
     1921 ctgcagcaga ccttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg
     1981 ttgatgaaat gatcagggaa gcagatatcg atggagaagg tcaggtaaac tacgaagagt
     2041 ttgtacaaat gatgacagcg aaggcggccg cagaacaaaa actcatctca gaagaggatc
     2101 tgaatggggc cgcaagcgag aaggacgagc tgtagtctag aagctcgctg atcagcctcg
     2161 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
     2221 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
     2281 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
     2341 tgggaagaca atagcaggca tgctggggat ggcccgggct ctatggcttc tgaggcggaa
     2401 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2461 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2521 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2581 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctctc ccccaaaaaa
     2641 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2701 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2761 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2821 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2881 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2941 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3001 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3061 ccctaactcc gcccagttcc gcccattctc cgcccctagg ctgactaatt ttttttattt
     3121 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3181 ttggaggcct aggcttttgc aaaaagctcc ccccccggga ggtccacaat ggttgaacaa
     3241 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg
     3301 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc
     3361 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactcca ggacgaggca
     3421 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc
     3481 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca
     3541 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat
     3601 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca
     3661 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg
     3721 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgta tgcccgacgg cgaggatctc
     3781 gtcgtgactc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct
     3841 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct
     3901 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac
     3961 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc
     4021 tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4081 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4141 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4201 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4261 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4321 atgtctgtat accgtcgatc tttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     4381 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     4441 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     4501 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4561 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4621 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4681 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4741 ttcggtgtag gtcgttcgct ccaagctggg ctgtatgcac gaaccccccg ttcagcccga
     4801 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4861 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4921 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4981 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5041 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5101 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5161 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5221 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5281 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     5341 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     5401 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     5461 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5521 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5581 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5641 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5701 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5761 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5821 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5881 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5941 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6001 cagttcgatg taacccactc gggcacccaa ctgatcttca gcatctttta ctttcaccag
     6061 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6121 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6181 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6241 tccgcgcaca tttccccgaa aagtgccacc tgacgtcaga tcgacggatc gggagatcg
//
