LOCUS       sequence-136827-        6114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent reporter of Orai3-associated calcium influx.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6114)
  AUTHORS   Dynes JL, Amcheslavsky A, Cahalan MD
  TITLE     Genetically targeted single-channel optical recording reveals 
            multiple Orai1 gating states and oscillations in calcium influx.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jan 12;113(2):440-5. doi: 
            10.1073/pnas.1523410113. Epub 2015 Dec 28.
  PUBMED    26712003
REFERENCE   2  (bases 1 to 6114)
  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"; doi: 
            "10.1073/pnas.1523410113"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2016-01-12- 12"; volume: "113"; issue: "2"; pages: "440-5"
COMMENT     Sequence Label: G-GECO1-Orai3
FEATURES             Location/Qualifiers
     source          1..6114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      600..609
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             621..677
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     primer_bind     890..911
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1278..1297)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             1413..1853
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     regulatory      2101..2110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2895..3016
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2932..2951)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2986..3005
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3023..3478)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3160..3181)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3372..3391
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3505..3609
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3611..3968
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3611..3802
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3634..3654)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3819..3954
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4003..4797
                     /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             4003..4797
                     /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(4057..4076)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4667..4686
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4985..5004)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5029..5076
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5405..5993
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5894..5913
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg
      601 ccaccatggt cgactcatca cgtcgtaagt ggaataagac aggtcacgca gtcagagcta
      661 taggtcggct gagctcactc gagaacgtct atatcaaggc cgacaagcag aagaacggca
      721 tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cggcgtgcag ctcgcctacc
      781 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
      841 tgagcgtgca gtccatactt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc
      901 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta
      961 ccggagggag catggtgagc aagggcgagg agctgttcac cggggtggtg cccatccagg
     1021 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggtgagggcg
     1081 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     1141 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     1201 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac atccaggagc
     1261 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1321 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1381 tcctggggca caagctggag tacaacacgc gtgaccaact gactgaagag cagatcgcag
     1441 aatttaaaga ggctttctcc ctatttgaca aggacgggga tgggacgata acaaccaagg
     1501 agctggggac ggtgatgcgg tctctggggc agaaccccac agaagcagag ctgcaggaca
     1561 tgatcaatga agtagatgcc gacggtgacg gcacaatcga cttccctgag ttcctgacaa
     1621 tgatggcaag aaaaatgaaa gacacagaca gtgaagaaga aattagagaa gcgttccgtg
     1681 tgtttgataa ggacggcaat ggctacatcg gcgcagcaga gcttcgccac gtgatgacaa
     1741 accttggaga gaagttaaca gatgaagagg ttgatgaaat gatcagggta gcagacatcg
     1801 atggggatgg tcaggtaaac tacgaagagt ttgtacaaat gatgacagcg aagcagatct
     1861 cgagctcaag cttcgaattc aagggcggcg agggggacgc gggcgagcag gccccgctga
     1921 accctgaggg cgagagccct gcaggctcgg ccacgtaccg ggagttcgtg caccgcggct
     1981 acctggacct catgggggcc agtcagcact cgctgcgggc gctcagctgg cgccgcctct
     2041 acctcagccg ggccaagctc aaagcttcca gccgcacgtc tgccttgctc tcgggcttcg
     2101 ccatggtggc catggtggag gtgcagctgg agagtgacca cgagtaccca ccaggcctgc
     2161 tggtggcctt cagtgcctgc accaccgtgc tggtggctgt gcacctcttt gcactcatgg
     2221 tctccacgtg tctgctgccc cacattgaag ctgtgagcaa catccacaac ctcaactctg
     2281 tccaccagtc gccacaccag agactgcacc gctacgtgga gctggcctgg ggcttctcca
     2341 ctgccctggg cacctttctc ttccttgctg aagttgtcct ggttggttgg gtcaagtttg
     2401 tgcccattgg ggctcccttg gacacaccga cccccatggt gcccacatcc cgggtgcccg
     2461 ggactctggc accagtggct acctccctta gtccagcttc caatctccca cggtcctctg
     2521 cgtctgcagc accgtcccag gctgagccag cctgcccacc ccggcaagcc tgtggtggtg
     2581 gtggggccca tgggccaggc tggcaagcag ccatggcctc cacagccatc atggtacccg
     2641 tggggctcgt gtttgtggcc tttgccctgc atttctaccg ctccttggtg gcacacaaga
     2701 cagaccgcta caagcaggaa ctagaggaac tgaatcgcct gcagggggag ctgcaggctg
     2761 tgtgaggatc caccggatct agataactga tcataatcag ccataccaca tttgtagagg
     2821 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     2881 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     2941 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3001 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3061 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3121 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3181 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3241 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3301 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3361 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     3421 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     3481 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3541 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3601 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     3661 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3721 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3781 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3841 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3901 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3961 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4021 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4081 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4141 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4201 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4261 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4321 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4381 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     4441 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     4501 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     4561 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     4621 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     4681 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     4741 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     4801 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     4861 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     4921 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     4981 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5041 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5101 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5161 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5221 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5281 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5341 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     5401 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     5461 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     5521 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     5581 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     5641 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     5701 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     5761 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     5821 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     5881 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     5941 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6001 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6061 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcattag ttat
//
