LOCUS       sequence-136141-        6132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent reporter of Orai1-associated calcium influx.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6132)
  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 6132)
  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-Orai1
FEATURES             Location/Qualifiers
     source          1..6132
                     /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      2176..2185
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2913..3034
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2950..2969)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3004..3023
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3041..3496)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3178..3199)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3390..3409
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3523..3627
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3629..3986
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3629..3820
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3652..3672)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3837..3972
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4021..4815
                     /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             4021..4815
                     /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(4075..4094)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4685..4704
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5003..5022)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5047..5094
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5423..6011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5912..5931
                     /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 catccggagc ccgccccgcc cccgagccgc agcagtcccg
     1921 agcttccccc aagcggcggc agcaccacca gcggcagccg ccggagccgc cgccgcagcg
     1981 gggacgggga gcccccgggg gccccgccac cgccgccgtc cgccgtcacc tacccggact
     2041 ggatcggcca gagttactcc gaggtgatga gcctcaacga gcactccatg caggcgctgt
     2101 cctggcgcaa gctctacttg agccgcgcca agcttaaagc ctccagccgg acctcggctc
     2161 tgctctccgg cttcgccatg gtggcaatgg tggaggtgca gctggacgct gaccacgact
     2221 acccaccggg gctgctcatc gccttcagtg cctgcaccac agtgctggtg gctgtgcacc
     2281 tgtttgcgct catgatcagc acctgcatcc tgcccaacat cgaggcggtg agcaacgtgc
     2341 acaatctcaa ctcggtcaag gagtcccccc atgagcgcat gcaccgccac atcgagctgg
     2401 cctgggcctt ctccaccgtc atcggcacgc tgctcttcct agctgaggtg gtgctgctct
     2461 gctgggtcaa gttcttgccc ctcaagaagc agccaggcca gccaaggccc accagcaagc
     2521 cccccgccag tggcgcagca gccaacgtca gcaccagcgg catcaccccg ggccaggcag
     2581 ctgccatcgc ctcgaccacc atcatggtgc ccttcggcct gatctttatc gtcttcgccg
     2641 tccacttcta ccgctcactg gttagccata agaccgaccg acagttccag gagctcaacg
     2701 agctggcgga gtttgcccgc ttacaggacc agctggacca cagaggggac caccccctga
     2761 cgcccggcag ccactatgcc tagggatcca ccggatctag ataactgatc ataatcagcc
     2821 ataccacatt tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc
     2881 tgaaacataa aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt
     2941 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3001 gttgtggttt gtccaaactc atcaatgtat cttaacgcgt aaattgtaag cgttaatatt
     3061 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     3121 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     3181 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     3241 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     3301 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     3361 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     3421 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     3481 ccgctacagg gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     3541 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     3601 cttcaataat attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt
     3661 gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3721 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     3781 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     3841 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     3901 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     3961 tttttggagg cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc
     4021 atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc
     4081 ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca
     4141 gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg
     4201 caagacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg
     4261 ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag
     4321 gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg
     4381 cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc
     4441 atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa
     4501 gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac
     4561 ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     4621 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     4681 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     4741 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     4801 gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc
     4861 tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg
     4921 ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg
     4981 cccaccctag ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg
     5041 ctatgacggc aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac
     5101 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
     5161 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
     5221 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac
     5281 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     5341 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     5401 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     5461 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     5521 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     5581 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     5641 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     5701 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     5761 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     5821 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     5881 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     5941 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6001 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6061 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca
     6121 tgcattagtt at
//
