LOCUS       sequence-136822-        6138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Green fluorescent reporter of mutant Orai1Y80E-associated calcium 
            influx.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6138)
  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 6138)
  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-GECO1.2-Orai1Y80E
FEATURES             Location/Qualifiers
     source          1..6138
                     /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"
     CDS             1419..1859
                     /codon_start=1
                     /product="calcium-binding protein"
                     /label=calmodulin
                     /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRSLGQNPTEA
                     ELQDMINEVDADGDGTIDFPEFLTMMARKMKDTDSEEEIREAFRVFDKDGNGYIGAAEL
                     RHVMTNLGEKLTDEEVDEMIRVADIDGDGQVNYEEFVQMMTAK"
     regulatory      2182..2191
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2919..3040
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2956..2975)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3010..3029
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3047..3502)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3184..3205)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3396..3415
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3529..3633
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3635..3992
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3635..3826
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3658..3678)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3843..3978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4027..4821
                     /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             4027..4821
                     /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(4081..4100)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4691..4710
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5009..5028)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5053..5100
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5429..6017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5918..5937
                     /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 cgacgagcag aagaacggca
      721 tcaaggcgta cttcaagatc cgccacaaca tcgagggcgg cggcgtgcag ctcgcctacc
      781 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
      841 tgagcgtgca gtccatgctt tcgaaagacc ccaacgagaa gcgcgatcac atggtcctgc
      901 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagggcggta
      961 ccggagggag cgaatccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     1021 tccaggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggtg
     1081 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     1141 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     1201 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc
     1261 aggagcgcac catcttcttc aagggcgacg gcaactacaa gacccgcgcc gaggtgaagt
     1321 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     1381 gcaacatcct ggggcacaag ctggagtaca acacgcgtga ccaactgact gaagagcaga
     1441 tcgcagaatt taaagaggct ttctccctat ttgacaagga cggggatggg acgataacaa
     1501 ccaaggagct ggggacggtg atgcggtctc tggggcagaa ccccacagaa gcagagctgc
     1561 aggacatgat caatgaagta gatgccgacg gtgacggcac aatcgacttc cctgagttcc
     1621 tgacaatgat ggcaagaaaa atgaaagaca cagacagtga agaagaaatt agagaagcgt
     1681 tccgtgtgtt tgataaggac ggcaatggct acatcggcgc agcagagctt cgccacgtga
     1741 tgacaaacct tggagagaag ttaacagatg aagaggttga tgaaatgatc agggtagcag
     1801 acatcgatgg ggatggtcag gtaaactacg aagagtttgt acaaatgatg acagcgaagc
     1861 agatctcgag ctcaagcttc gaattccatc cggagcccgc cccgcccccg agccgcagca
     1921 gtcccgagct tcccccaagc ggcggcagca ccaccagcgg cagccgccgg agccgccgcc
     1981 gcagcgggga cggggagccc ccgggggccc cgccaccgcc gccgtccgcc gtcacctacc
     2041 cggactggat cggccagagt tactccgagg tgatgagcct caacgagcac tccatgcagg
     2101 cgctgtcctg gcgcaagctc gagttgagcc gcgccaagct taaagcctcc agccggacct
     2161 cggctctgct ctccggcttc gccatggtgg caatggtgga ggtgcagctg gacgctgacc
     2221 acgactaccc accggggctg ctcatcgcct tcagtgcctg caccacagtg ctggtggctg
     2281 tgcacctgtt tgcgctcatg atcagcacct gcatcctgcc caacatcgag gcggtgagca
     2341 acgtgcacaa tctcaactcg gtcaaggagt ccccccatga gcgcatgcac cgccacatcg
     2401 agctggcctg ggccttctcc accgtcatcg gcacgctgct cttcctagct gaggtggtgc
     2461 tgctctgctg ggtcaagttc ttgcccctca agaagcagcc aggccagcca aggcccacca
     2521 gcaagccccc cgccagtggc gcagcagcca acgtcagcac cagcggcatc accccgggcc
     2581 aggcagctgc catcgcctcg accaccatca tggtgccctt cggcctgatc tttatcgtct
     2641 tcgccgtcca cttctaccgc tcactggtta gccataagac cgaccgacag ttccaggagc
     2701 tcaacgagct ggcggagttt gcccgcttac aggaccagct ggaccacaga ggggaccacc
     2761 ccctgacgcc cggcagccac tatgcctagg gatccaccgg atctagataa ctgatcataa
     2821 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2881 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2941 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3001 attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt
     3061 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     3121 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     3181 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     3241 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     3301 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     3361 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     3421 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     3481 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3541 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3601 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3661 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3721 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3781 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3841 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3901 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3961 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     4021 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     4081 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     4141 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     4201 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     4261 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     4321 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     4381 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     4441 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     4501 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4561 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4621 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4681 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4741 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4801 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4861 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4921 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4981 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     5041 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     5101 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     5161 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     5221 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     5281 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5341 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5401 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5461 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5521 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5581 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5641 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5701 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5761 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5821 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5881 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5941 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6001 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6061 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6121 ccgccatgca ttagttat
//
