LOCUS       sequence-311156-        6238 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cyan-fluorescent homoFRET/anisotropy-based genetically encoded 
            calcium indicator, targeted to endoplasmic reticulum lumen..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6238)
  AUTHORS   Ross BL, Tenner B, Markwardt ML, Zviman A, Shi G, Kerr JP, Snell NE,
            McFarland JJ, Mauban JR, Ward CW, Rizzo MA, Zhang J
  TITLE     Single-color, ratiometric biosensors for detecting signaling 
            activities in live cells.
  JOURNAL   eLife. 2018 Jul 3;7. pii: 35458. doi: 10.7554/eLife.35458
  PUBMED    29968564
REFERENCE   2  (bases 1 to 6238)
  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: "eLife."; 
            date: "2018-07-3"; volume: "7. pii: 35458. doi"; pages: " 
            10.7554/eLife.3545"
COMMENT     Sequence Label: pEGFPC1-Cerulean3-Cerulean3-FLARE-D1ER
FEATURES             Location/Qualifiers
     source          1..6238
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        57..360
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        361..564
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     515..535
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      603..612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             609..701
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="MDSKGSSQKGSRLLLLLVVSNLLLCQGVVST"
     regulatory      711..720
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1239..1258)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2034..2123
                     /codon_start=1
                     /product="signal sequence from bovine prolactin"
                     /label=prolactin signal sequence
                     /translation="DSKGSSQKGSRLLLLLVVSNLLLCQGVVST"
     polyA_signal    3022..3143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3059..3078)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3113..3132
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3150..3605)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3287..3308)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3499..3518
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3632..3736
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3738..4095
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3738..3929
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3761..3781)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3946..4081
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4130..4924
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4130..4924
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4184..4203)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4794..4813
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5112..5131)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5156..5203
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5532..6120
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6021..6040
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
       61 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
      121 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
      181 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
      241 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
      301 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
      361 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
      421 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      481 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      541 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcggcaaagc
      601 ttgccaccat ggacagcaaa ggttcgtcgc agaaagggtc ccgcctgctc ctgctgctgg
      661 tggtgtcaaa tctactcttg tgccagggtg tggtctccac cggaccggtc gccaccatgg
      721 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
      781 tgctgctgcc cgacaaccac tacctgagca cccagtccaa gctgagcaaa gaccccaacg
      841 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
      901 tggacgagct gtacaagggt ggcagcggtg gcatggtgag caagggcgag gagctgttca
      961 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1021 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatcagca
     1081 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgagc tggggcgtgc
     1141 agagcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1201 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1261 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1321 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacgcc atccacggca
     1381 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc ggcctcaact
     1441 gcaacatcga gtccggactc agatctcgag ctcaagcttc taattctgca gcccgcatgc
     1501 atgaccaact gacagaagag cagattgcaa aattcaaaga agccttctca ttattcgaca
     1561 aggatgggga cggcaccatc accacaaagg aacttggcac cgttatgagg tcgcttggac
     1621 aaaacccaac ggaagcagaa ttgcaggata tgatcaatga agtcgatgct gatggcaatg
     1681 gaacgattta ctttcctgaa tttcttacta tgatggctag aaaaatgaag gacacagaca
     1741 gcgaagagag aatccgaaaa gcattccgtg tttttgacaa ggatgggaac ggctacatca
     1801 gcgctgctga attacgtcac gtcatgacaa acctcgggga gaagttaaca gatgaagaag
     1861 ttgatgaaat gataagggaa gcagatatcg atggtgatgg ccaagtaaac tatgaagagt
     1921 ttgtacaaat gatgacagca aaggggggga agagggagtg ggagaaaaac ttcattgccg
     1981 tcagcgctgc caacgagttc gagaagatct ccagctccgg ggcactggag ctcgacagca
     2041 aaggttcgtc gcagaaaggg tcccgcctgc tcctgctgct ggtggtgtca aatctactct
     2101 tgtgccaggg tgtggtctcc accggaccag tcgtgtcaaa gggcgaagag ctgtttactg
     2161 gagtcgtccc aattctggtg gagctggatg gagatgtcaa cgggcataag ttttcagtgc
     2221 gaggagaggg agaaggagac gctaccaacg gcaagctgac actgaaattc atcagcacca
     2281 cagggaagct gcccgtgcct tggccaaccc tggtcactac cctgagttgg ggcgtccagt
     2341 catttgcccg atacccagac cacatgaagc agcatgattt ctttaaatcc gctatgcccg
     2401 agggctacgt gcaggaacgg acaattttct ttaaggacga tggaacatac aaaactagag
     2461 cagaggtgaa gttcgaagga gacactctgg tcaacaggat cgagctgaag ggcattgact
     2521 ttaaagaaga tggaaatatc ctgggccaca aactggagta caacgccatc catggcaacg
     2581 tgtacattac agctgataag cagaaaaacg ggattaaggc aaatttcgga ctgaactgca
     2641 atgtggaaga cgggagtgtc cagctggccg atcactacca gcagaacact cctatcggcg
     2701 acgggcccgt gctgctgcct gataatcatt atctgagcac ccagtccgtc ctgtctaagg
     2761 accccaatga gaaaagagat cacatggtgc tgctggagtt tgtgaccgct gctggcatta
     2821 ccctggggat ggacgagctg tacaagaagg acgagctgta agaattctgc agtcgacggt
     2881 accgcgggcc cgggatccac cggatctaga taactgatca taatcagcca taccacattt
     2941 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa
     3001 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     3061 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3121 tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt tgttaaaatt
     3181 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3241 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3301 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3361 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3421 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3481 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3541 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     3601 cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     3661 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     3721 ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg
     3781 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3841 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3901 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3961 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     4021 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     4081 ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca
     4141 agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg
     4201 ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg
     4261 cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc
     4321 agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt
     4381 cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc
     4441 atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca
     4501 tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc
     4561 acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg
     4621 gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct
     4681 cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc
     4741 tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc
     4801 tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta
     4861 cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt
     4921 ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga
     4981 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
     5041 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg
     5101 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     5161 ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg
     5221 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     5281 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     5341 aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat
     5401 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     5461 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     5521 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     5581 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     5641 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     5701 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     5761 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     5821 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     5881 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     5941 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6001 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6061 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6121 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     6181 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagt
//
