LOCUS       sequence-462300-        6169 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 6169)
  AUTHORS   Dynes JL, Yeromin AV, Cahalan MD
  TITLE     Photoswitching alters fluorescence readout of jGCaMP8 Ca(2+) 
            indicators tethered to Orai1 channels.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Sep 26;120(39):e2309328120. doi: 
            10.1073/pnas.2309328120. Epub 2023 Sep 20.
  PUBMED    37729200
REFERENCE   2  (bases 1 to 6169)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2023-09-26"; volume: "120(39):e2309328120. 
            doi"; pages: " 10.1073/pnas.2309328120. Epub 2023 Sep 20"
COMMENT     Sequence Label: Orai1-jGCaMP8f
FEATURES             Location/Qualifiers
     source          1..6169
                     /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      922..931
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1865..1886
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1992..2013)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2253..2272)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2957..3078
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2994..3013)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3048..3067
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3085..3540)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3222..3243)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3434..3453
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3567..3671
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3673..4030
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3673..3864
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3696..3716)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3881..4016
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4065..4859
                     /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             4065..4859
                     /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(4119..4138)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4729..4748
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5047..5066)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5091..5138
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5467..6055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5956..5975
                     /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 gctagcgcta
      601 ccggactcag atctcgagcc accatgcatc cggagcccgc cccgcccccg agccgcagca
      661 gtcccgagct tcccccaagc ggcggcagca ccaccagcgg cagccgccgg agccgccgcc
      721 gcagcgggga cggggagccc ccgggggccc cgccaccgcc gccgtccgcc gtcacctacc
      781 cggactggat cggccagagt tactccgagg tgatgagcct caacgagcac tccatgcagg
      841 cgctgtcctg gcgcaagctc tacttgagcc gcgccaagct taaagcctcc agccggacct
      901 cggctctgct ctccggcttc gccatggtgg caatggtgga ggtgcagctg gacgctgacc
      961 acgactaccc accggggctg ctcatcgcct tcagtgcctg caccacagtg ctggtggctg
     1021 tgcacctgtt tgcgctcatg atcagcacct gcatcctgcc caacatcgag gcggtgagca
     1081 acgtgcacaa tctcaactcg gtcaaggagt ccccccatga gcgcatgcac cgccacatcg
     1141 agctggcctg ggccttctcc accgtcatcg gcacgctgct cttcctagct gaggtggtgc
     1201 tgctctgctg ggtcaagttc ttgcccctca agaagcagcc aggccagcca aggcccacca
     1261 gcaagccccc cgccagtggc gcagcagcca acgtcagcac cagcggcatc accccgggcc
     1321 aggcagctgc catcgcctcg accaccatca tggtgccctt cggcctgatc tttatcgtct
     1381 tcgccgtcca cttctaccgc tcactggtta gccataagac cgaccgacag ttccaggagc
     1441 tcaacgagct ggcggagttt gcccgcttac aggaccagct ggaccacaga ggggaccacc
     1501 ccctgacgcc cggcagccac tatgcccgaa ttctgcagtc gacggtaccg cgggcccggg
     1561 atccaccggt cgccaccctg cacacgcgtc gcaagaagac cttcaaggag gtggccaccg
     1621 ccgtgaagat catcgcccgc ctgatgggcc tgaagatcaa cgtctatatc aaggccgaca
     1681 agcagaagaa cggcatcaag gcgaacttcc acatccgcca caacatcgag gacggcggcg
     1741 tgcagctcgc ctaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1801 ccgacaacca ctacctgagc gtggagtcca aactttcgaa agaccccaac gagaagcgcg
     1861 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1921 tgtacaaggg cggtaccgga gggagcatgg tgagcaaggg cgaggagctg ttcaccgggg
     1981 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2041 gcgagggtga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2101 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2161 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2221 gctacatcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2281 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2341 aggaggacgg caacatcctg gggcacaagc tggagtacaa cctgccggac caactgactg
     2401 aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac ggggatggga
     2461 caataacaac caaggagctg gggacggtga tgcggtctct cggactgaac cccacagaag
     2521 cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca atcgacttcc
     2581 ctgagttcct gacaatgatg gcaagaaaaa tgaaatacag ggacacggaa gaagaaatta
     2641 gagaagcgtt cggtgtgttt gataaggatg gcaatggcta catcagtgca gcagagcttc
     2701 gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat gaaatgatca
     2761 gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta caaatgatga
     2821 cagcgaagct ttaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     2881 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2941 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     3001 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     3061 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     3121 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     3181 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     3241 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     3301 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3361 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3421 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3481 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3541 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3601 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3661 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3721 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3781 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3841 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3901 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3961 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4021 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4081 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4141 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4201 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     4261 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4321 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4381 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4441 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4501 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4561 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4621 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4681 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4741 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4801 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4861 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4921 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4981 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     5041 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     5101 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     5161 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     5221 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     5281 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     5341 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5401 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5461 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5521 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5581 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     5641 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5701 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5761 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5821 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5881 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5941 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6001 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6061 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     6121 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
