LOCUS       sequence-117195-        7165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of ORAI1-hBACCS2, a fusion protein of ORAI1 and
            human blue light-activated Ca2+ channel switch.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7165)
  AUTHORS   Ishii T, Sato K, Kakumoto T, Miura S, Touhara K, Takeuchi S, Nakata 
            T
  TITLE     Light generation of intracellular Ca(2+) signals by a genetically 
            encoded protein BACCS.
  JOURNAL   Nat Commun. 2015 Aug 18;6:8021. doi: 10.1038/ncomms9021.
  PUBMED    26282514
REFERENCE   2  (bases 1 to 7165)
  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: "2015-08-18"; volume: "6:8021. doi"; pages: " 
            10.1038/ncomms9021"
COMMENT     Sequence Label: ORAI1-hBACCS2
FEATURES             Location/Qualifiers
     source          1..7165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      909..918
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1565..1591
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1598..1624
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1631..1657
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    3224..3812
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3277..3297)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3695..3706)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3953..4074
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3990..4009)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4044..4063
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4081..4536)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4218..4239)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4430..4449
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4563..4667
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4669..5026
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4669..4860
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4692..4712)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4877..5012
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5061..5855
                     /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             5061..5855
                     /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(5115..5134)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5725..5744
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6043..6062)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6087..6134
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6463..7051
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6952..6971
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgctc
      601 tcgagccacc atgcatccgg agcccgcccc gcccccgagc cgtagcagtc ccgagcttcc
      661 cccaagcggc ggcagcacca ccagcggcag ccgccggagc cgccgccgca gcggggacgg
      721 ggagcccccg ggggccccgc caccgccgcc gtccgccgtc acctacccgg actggatcgg
      781 ccagagttac tccgaggtga tgagcctcaa cgagcactcc atgcaggcgc tgtcctggcg
      841 caagctctac ttgagccgcg ccaagcttaa agcctccagc cggacctcgg ctctgctctc
      901 cggcttcgcc atggtggcaa tggtggaggt gcagctggac gctgaccacg actacccacc
      961 ggggctgctc atcgccttca gtgcctgcac cacagtgctg gtggctgtgc acctgtttgc
     1021 gctcatgatc agcacctgca tcctgcccaa catcgaggcg gtgagcaacg tgcacaatct
     1081 caactcggtc aaggagtccc cccatgagcg catgcaccgc cacatcgagc tggcctgggc
     1141 cttctccacc gtcattggca cgctgctctt cctagctgag gtggtgctgc tctgctgggt
     1201 caagttcttg cccctcaaga agcagccagg ccagccaagg cccaccagca agccccccgc
     1261 cagtggcgca gcagccaacg tcagcaccag cggcatcacc ccgggccagg cagctgccat
     1321 cgcctcgacc accatcatgg tgcccttcgg cctgatcttt atcgtcttcg ccgtccactt
     1381 ctaccgctca ctggttagcc ataagaccga ccgacagttc caggagctca acgagctggc
     1441 ggagtttgcc cgcttacagg accagctgga ccacagaggg gaccaccccc tgacgcccgg
     1501 cagccactat gcccgaattc tccagtccac ggtgccacgg gccagagatc caccggtcgg
     1561 aggatacccc tatgacgttc cagattatgc tggaagctat ccatacgatg tccccgacta
     1621 cgctggctcc tatccctacg atgtgccaga ctatgcccca ccggtcggag gaagcggtgg
     1681 atctttggct actacacttg aacgtattga gaagaacttt gtcattactg acccaagatt
     1741 gccagataat cccattatat tcgcgtccga tagtttcttg cagttgacag aatatagccg
     1801 tgaagaaatt ttgggaagaa actgcaggtt tctacaaggt cctgaaactg atcgcgcgac
     1861 agtgagaaaa attagagatg ccatagataa ccaaacagag gtcactgttc agctgattaa
     1921 ttatacaaag agtggtaaaa agttctggaa cctctttcac ttgcagccta tgcgagatca
     1981 gaagggagat gtccagtact ttattggggt tcagttggat ggaactgagc atgtccgaga
     2041 tgctgccgag agagagggag tcatgctgat taagaaaact gcagaaaatc ttcagaagtg
     2101 gctgcagctg acacatgagg tggaggtgca atattacaac atcaagaagc aaaatgctga
     2161 gaagcagctg ctggtggcca aggagggggc tgagaagata aaaaagaaga gaaacacact
     2221 ctttggcacc ttccacgtgg cccacagctc ttccctggat gatgtagatc ataaaattct
     2281 aacagctaag caagcactga gcgaggtgac agcagcattg cgggagcgcc tgcaccgctg
     2341 gcaacagatc gagatcctct gtggcttcca gattgtcaac aaccctggca tccacggagg
     2401 gtcaggagga agtggaggag gtatcctgca gtctcggggt ggcagtggtg gttcaggcgg
     2461 ttctggatct ttggctacta cacttgaacg tattgagaag aactttgtca ttactgaccc
     2521 aagattgcca gataatccca ttatattcgc gtccgatagt ttcttgcagt tgacagaata
     2581 tagccgtgaa gaaattttgg gaagaaactg caggtttcta caaggtcctg aaactgatcg
     2641 cgcgacagtg agaaaaatta gagatgccat agataaccaa acagaggtca ctgttcagct
     2701 gattaattat acaaagagtg gtaaaaagtt ctggaacctc tttcacttgc agcctatgcg
     2761 agatcagaag ggagatgtcc agtactttat tggggttcag ttggatggaa ctgagcatgt
     2821 ccgagatgct gccgagagag agggagtcat gctgattaag aaaactgcag aaaatcttca
     2881 gaagtggctg cagctgacac atgaggtgga ggtgcaatat tacaacatca agaagcaaaa
     2941 tgctgagaag cagctgctgg tggccaagga gggggctgag aagataaaaa agaagagaaa
     3001 cacactcttt ggcaccttcc acgtggccca cagctcttcc ctggatgatg tagatcataa
     3061 aattctaaca gctaagcaag cactgagcga ggtgacagca gcattgcggg agcgcctgca
     3121 ccgctggcaa cagatcgaga tcctctgtgg cttccagatt gtcaacaacc ctggcatcca
     3181 ctgagtcgac ggtaccgcgg gcccgggatc taagcttatc gataatcaac ctctggatta
     3241 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     3301 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     3361 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     3421 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     3481 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     3541 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     3601 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     3661 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     3721 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3781 gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtggatcca ccggatctag
     3841 ataactgatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3901 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3961 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4021 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaacgcgt
     4081 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     4141 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     4201 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     4261 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     4321 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     4381 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     4441 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     4501 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4561 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4621 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4681 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4741 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4801 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4861 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4921 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4981 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     5041 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     5101 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5161 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5221 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     5281 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5341 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5401 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5461 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5521 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5581 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5641 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5701 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5761 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5821 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5881 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5941 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     6001 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     6061 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     6121 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     6181 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     6241 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     6301 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6361 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6421 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6481 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6541 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6601 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6661 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6721 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6781 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6841 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6901 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6961 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7021 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7081 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     7141 ggataaccgt attaccgcca tgcat
//
