LOCUS       sequence-492867-        6653 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bioluminescent endolysosomal Ca2+ indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6653)
  AUTHORS   Calvo B, Torres-Vidal P, Delrio-Lorenzo A, Rodriguez C, Aulestia FJ,
            Rojo-Ruiz J, Callejo B, McVeigh BM, Keller M, Grimm C, Oorschot V, 
            Moiseenkova-Bell V, Yule DI, Garcia-Sancho J, Patel S, Alonso MT
  TITLE     Direct measurements of luminal Ca2+ with endo-lysosomal GFP-aequorin
            reveal functional IP3 receptors.
  JOURNAL   J Cell Biol. 2025 Dec 1;224(12):e202410094. doi: 
            10.1083/jcb.202410094. Epub 2025 Nov 7.
  PUBMED    41201441
REFERENCE   2  (bases 1 to 6653)
  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: "J Cell 
            Biol."; date: "2025-12-1"; volume: "224(12):e202410094. doi"; pages:
            " 10.1083/jcb.202410094. Epub 2025 Nov 7"
COMMENT     Sequence Label: pHSV-ELGA
FEATURES             Location/Qualifiers
     source          1..6653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     154..171
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    186..326
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(252..271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     412..434
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     regulatory      1506..1515
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2178..2888
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEQLFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2217..2238)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2478..2497)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2825..2846
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3881..4015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3918..3937)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3972..3991
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4843..4861)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4929..5033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5034..5102,5103..5894)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5252..5271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6065..6653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6554..6573
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
      241 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
      301 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
      361 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
      421 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagcat gaattattcc
      481 attatgcacg accccgcccc gacgccggca cgccgggggc ccgtggccgc ggcccgttgg
      541 tcgaaccccc ggccccgccc atccgcgcca tctgccatgg gcggggcgcg agggcgggtg
      601 ggtccgcgcc ccgccccgca tggcatctca ttaccgcccg atccggcggt ttccgcttcc
      661 gttccgcatg ctaacgagga acaggcaggg ggcggggccc gggccccgac ttcccggttc
      721 ggcggtaatg agatacgagc cccgcgcgcc cgttggccgt ccccgggccc cccggtcccg
      781 cccgccggac gccgggacca acgggacggc gggcggccca agggccgccc gccttgccgc
      841 ccccccattg gccggcgggc gggaccgccc caagggggcg gggccgccgg gtaaaagaag
      901 tgagaacgcg aagcgttcgc acttcgtccc aatatatata tattattagg gcgaagtgcg
      961 agcactggcg ccgtgcccga ctccgcgccg gccccggggg cgggcccggg cggcgggggg
     1021 cgggtctctc cggcgcacat aaaggcccgg cgcgaccgac gcccgcagac ggcgccggcc
     1081 acgaacgacg ggagcggctg cggagcacgc ggaccgggag cgggagtcgc agagggccgt
     1141 cggagcggac ggcgtcggca tcgcgacgcc ccggctcggg atcgggatcg catcggaaag
     1201 ggacacgcgg acgcgggggg gaaagacccg cccaccccac ccacgaaaca caggggacgc
     1261 accccggggg cctccgacga cagaaaccca ccggtccgcc ttttttgcac gggtaagcac
     1321 cttgggtggg cggaggaggg ggggacgcgg gggcggagga ggggggacgc gggggcggag
     1381 gaggggggac gcgggggcgg aggagggggg acgcgggggc ggaggagggg ggacgcgggg
     1441 gcggaggagg gggctcaccc gcgttcgtgc cttcccgcag gaggaacgtc ctcgtcgata
     1501 agcttgccac catggcgatt ctttttgctg ttgttgccag ggggaccact atccttgcca
     1561 aacatgcttg gtgtggagga aacttcctgg aggtgacaga gcagattctg gctaagatac
     1621 cttctgaaaa taacaaacta acgtactcac atggcaatta tttgtttcat tacatctgcc
     1681 aagacaggat tgtatatctt tgtatcactg atgatgattt tgaacgttcc cgagccttta
     1741 attttctgaa tgagataaag aagaggttcc agactactta cggttcaaga gcacagacag
     1801 cacttccata tgccatgaat agcgagttct caagtgtctt agctgcacag ctgaagcatc
     1861 actctgagaa taagggccta gacaaagtga tggagactca agcccaagtg gatgaactga
     1921 aaggaatcat ggtcagaaac atagatctgg tagctcagcg aggagaaaga ttggaattat
     1981 tgattgacaa aacagaaaat cttgtggatt cttctgtcac cttcaaagct accagcagaa
     2041 atcttgctcg agccatgtgt atgaagaacc tcaagctcac tattatcatc atcatcgtat
     2101 caattgtgtt catctatatc attgtttcac ctctctgtgg tggatttaca tggccaagct
     2161 gtgtgaagaa aaagcttagc aagggcgagc agctgttcac cggggtggtg cccatcctgg
     2221 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     2281 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     2341 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     2401 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     2461 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     2521 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     2581 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     2641 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcagcg
     2701 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     2761 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     2821 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactcacggc atggacgagc
     2881 tgtacaagtc cggcgggagc ggatccggcg gccagtccgg actcagatct gtcaaactta
     2941 catcagactt cgacaaccca agatggattg gacgacacaa gcatatgttc aatttccttg
     3001 atgtcaacca caatggaaaa atctctcttg acgagatggt ctacaaggca tctgatattg
     3061 tcatcaataa ccttggagca acacctgagc aagccaaacg acacaaagat gctgtggaag
     3121 ccttcttcgg aggagctgga atgaaatatg gtgtggaaac tgattggcct gcatatattg
     3181 aaggatggaa aaaattggct actgatgaat tggagaaata cgccaaaaac gaaccaaccc
     3241 tcatccgcat ctggggtgat gctttgtttg atatcgtcga caaagcccaa aatggagcca
     3301 ttacactgga tgaatggaaa gcatacacca aagctgctgg tatcatccaa tcatcagaag
     3361 attgcgagga aacattcaga gtgtgcgata ttgatgaaag tggacaactc gatgttgatg
     3421 agatgacaag acaacattta ggattttggt acaccatgga tcctgcttgc gaaaagctct
     3481 acggtggagc tgtcccctaa gaagctctac ggtggtgatg caccctagga agatgatgtg
     3541 attttgaata aaacactgat gaattccagc tgagcgccgg tcgctaccat taccagttgg
     3601 tctggtgtca aaaataataa taaccgggca ggccatgtct gcccgtattt cgcgtaagga
     3661 aatccattat gtactattta aaaaacacaa acttttggat gttcggttta ttctttttct
     3721 tttacttttt tatcatggga gcctacttcc cgtttttccc gatttggcta catgacatca
     3781 accatatcag caaaagtgat acgggtatta tttttgccgc tatttctctg ttctcgctat
     3841 tattccaacc gctgtttggt ctgctttctg acaaactcgg aacttgttta ttgcagctta
     3901 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     3961 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctctga
     4021 cctgagattg gcggcactga ggtagagatg cccgaacccc cccgagggag cgcgggacgc
     4081 ggccggggag ggctggggcc ggggagggct ggggccgggg agggctgggg ccggggaggg
     4141 ctggggccgg ggagggctgg ggccggggag ggctggggcc ggggagggct ggggctgggg
     4201 aggggcggtg gtgtgtagca ggagcggtgt gttgcgccgg ggtacgtctg gaggagcggg
     4261 aggtgcgcgg tgacgtgtgg atgaggaaca ggagttgttg cgcggtgagt tgtcgctgtg
     4321 agttgtgttg ttgggcaggt gtggtggatg acgtgacgtg tgacgtgcgg agtgcgccgt
     4381 gctctgttgg tttcacctgt ggggagcggg gggaggagcg gggggaggag cggggggagg
     4441 agcgggggga ggagcggggg gaggagcggg gggaggagcg gggggaggag cggggggagg
     4501 agcgggggga ggagcggggg gaggagcggc cagaccccga aaacgggccc cccccaaaac
     4561 acaccccccg ggggtcgcgc gcggcccttt aaagcggcgg cggcgggcag cccgggcccc
     4621 ccgcggccga gactagcgag ttagacaggc aagcactact cgcctctgca cgcacatgct
     4681 tgcctgtcaa actctaccac cccggcacgc tctctgtctc catggcccgc cgccgccgcc
     4741 atcgcggccc ccgccgcccc cggccgcccg ggcccacggg cgccgtccca accgcacagt
     4801 cccaggtaac gactctagga attaattctt gaagacgaaa gggcctcgtg atacgcctat
     4861 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4921 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4981 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5041 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5101 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5161 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5221 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     5281 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5341 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5401 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5461 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5521 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     5581 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5641 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5701 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5761 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5821 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5881 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     5941 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6001 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6061 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6121 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6181 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6241 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6301 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6361 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6421 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6481 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6541 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6601 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaa
//
