LOCUS       sequence-150677-        7609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GCaMP6 is a genetically encoded fluorescent Ca2+ sensor . 
            TPC2-GCaMP6 allows the visualization of Ca2+ release from 
            intracellular Ca2+ stores in real time.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7609)
  AUTHORS   Ambrosio AL, Boyle JA, Di Pietro SM
  TITLE     TPC2 mediates new mechanisms of platelet dense granule membrane 
            dynamics through regulation of Ca2+ release.
  JOURNAL   Mol Biol Cell. 2015 Sep 15;26(18):3263-74. doi: 
            10.1091/mbc.E15-01-0058. Epub 2015 Jul 22.
  PUBMED    26202466
REFERENCE   2  (bases 1 to 7609)
  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: 
            "10.1091/mbc.E15-01-0058"; journalName: "Mol Biol Cell"; date: 
            "2015-09-15- 15"; volume: "26"; issue: "18"; pages: "3263-74"
COMMENT     Sequence Label: pGCaMP6m-N3-TPC2
FEATURES             Location/Qualifiers
     source          1..7609
                     /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"
     CDS             join(2923..2931,2932..2949,2950..2952,2953..2985,2986..2988,
                     2989..3012,3013..3039,3040..3096,3097..3102,3103..3372,
                     3373..3390,3391..3825,3826..3831,3832..4275)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6m
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      3019..3028
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     3309..3330
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3436..3457)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3697..3716)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    4397..4518
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4434..4453)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4488..4507
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4525..4980)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4662..4683)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4874..4893
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5007..5111
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5113..5470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5113..5304
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5136..5156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5321..5456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5505..6299
                     /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             5505..6299
                     /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(5559..5578)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6169..6188
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6487..6506)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6531..6578
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6907..7495
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7396..7415
                     /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 atctcgagct caagcttatg gcggaacccc aggcggagtc ggagcccctg
      661 ctgggcgggg cccgcggcgg tggcggcgac tggccggcgg ggctgaccac ttaccgcagc
      721 atccaagtcg gccctggtgc cgcggccagg tgggacctct gcattgatca ggctgtggtc
      781 ttcatcgaag atgctattca gtaccgctcc atcaaccacc gggtggatgc cagctcgatg
      841 tggctttacc gacggtatta ctcgaacgta tgccaacgga ctttgagctt caccatcttc
      901 ttgatcctgt ttttggcttt tatcgagacc ccatcctcac tcaccagcac ggcggacgtg
      961 cgctaccgcg ctgccccctg ggagccgccc tgcggcctga ccgagagtgt cgaggtgctc
     1021 tgcctgctgg tctttgcggc cgacctctct gtgaagggtt acctgttcgg gtgggcccat
     1081 ttccagaaaa acctttggct gctgggctac ctcgtggtgc tggtggtgtc tctggtggac
     1141 tggaccgtgt ccctgagtct cgtgtgtcat gagcccctgc ggatccgccg gcttctccgt
     1201 cccttcttcc tgctgcagaa ctcctctatg atgaagaaga ccttgaaatg catccgctgg
     1261 tcgctgccgg aaatggccag cgtcgggctg ctgctggcca tccacctgtg cctcttcacc
     1321 atgttcggaa tgctgctgtt cgctggtggg aagcaggatg atgggcagga cagggagagg
     1381 ctgacctact tccagaacct gcctgagtct ctgacttccc tcctggtgct gctgaccacg
     1441 gccaacaacc ccgatgtgat gattcctgcg tattccaaga accgggccta tgccatcttc
     1501 ttcatagtct tcactgtgat aggaagcctg tttctgatga acctgctgac agccatcatc
     1561 tacagtcagt tccggggcta cctgatgaaa tctctccaga cctcgctgtt tcggaggcgg
     1621 ctgggaaccc gggctgcctt tgaagtccta tcctccatgg tgggggaggg aggagccttc
     1681 cctcaggcag ttggggtgaa gccccagaac ttgctgcagg tgcttcagaa ggtccagctg
     1741 gacagctccc acaaacaggc catgatggag aaggtgcgtt cctacggcag tgttctgctg
     1801 tcagctgagg agtttcagaa gctcttcaac gagcttgaca gaagtgtggt taaagagcac
     1861 ccgccgaggc ccgagtacca gtctccgttt ctgcagagcg cccagttcct cttcggccac
     1921 tactactttg actacctggg gaacctcatc gccctggcaa acctggtgtc catttgcgtg
     1981 ttcctggtgc tggatgcaga tgtgctgcct gctgagcgtg atgacttcat cctggggatt
     2041 ctcaactgcg tcttcattgt gtactacctg ttggagatgc tgctcaaggt ctttgccctg
     2101 ggcctgcgag ggtacctgtc ctaccccagc aacgtgtttg acgggctcct caccgttgtc
     2161 ctgctggttt tggagatctc aactctggct gtgtaccgat tgccacaccc aggctggagg
     2221 ccggagatgg tgggcctgct gtcgctgtgg gacatgaccc gcatgctgaa catgctcatc
     2281 gtgttccgct tcctgcgtat catccccagc atgaagccga tggccgtggt ggccagtacc
     2341 gtcctgggcc tggtgcagaa catgcgtgct tttggcggga tcctggtggt ggtctactac
     2401 gtatttgcca tcattgggat caacttgttt agaggcgtca ttgtggctct tcctggaaac
     2461 agcagcctgg cccctgccaa tggctcggcg ccctgtggga gcttcgagca gctggagtac
     2521 tgggccaaca acttcgatga ctttgcggct gccctggtca ctctgtggaa cttgatggtg
     2581 gtgaacaact ggcaggtgtt tctggatgca tatcggcgct actcaggccc gtggtccaag
     2641 atctattttg tattgtggtg gctggtgtcg tctgtcatct gggtcaacct gtttctggcc
     2701 ctgattctgg agaacttcct tcacaagtgg gacccccgca gccacctgca gccccttgct
     2761 gggaccccag aggccaccta ccagatgact gtggagctcc tgttcaggga tattctggag
     2821 gagcccgagg aggatgagct cacagagagg ctgagccagc acccgcacct gtggctgtgc
     2881 agggagaatt ctgcagtcga cggtaccgcg ggcccgggat ccatgggttc tcatcatcat
     2941 catcatcatg gtatggctag catgactggt ggacagcaaa tgggtcggga tctgtacgac
     3001 gatgacgata aggatctcgc caccatggtc gactcatcac gtcgtaagtg gaataagaca
     3061 ggtcacgcag tcagagctat aggtcggctg agctcactcg agaacgtcta tatcaaggcc
     3121 gacaagcaga agaacggcat caaggcgaac ttcaagatcc gccacaacat cgaggacggc
     3181 ggcgtgcagc tcgcctacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3241 ctgcccgaca accactacct gagcgtgcag tccaaacttt cgaaagaccc caacgagaag
     3301 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3361 gagctgtaca agggcggtac cggagggagc atggtgagca agggcgagga gctgttcacc
     3421 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
     3481 tccggcgagg gtgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
     3541 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
     3601 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
     3661 gaaggctaca tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
     3721 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     3781 ttcaaggagg acggcaacat cctggggcac aagctggagt acaacctgcc ggaccaactg
     3841 actgaagagc agatcgcaga atttaaagag gctttctccc tatttgacaa ggacggggat
     3901 gggacaataa caaccaagga gctggggacg gtgatgcggt ctctggggca gaaccccaca
     3961 gaagcagagc tgcaggacat gatcaatgaa gtagatgccg acggtgacgg cacaatcgac
     4021 ttccctgagt tcctgacaat gatggcaaga aaagggagct acagggacac ggaagaagaa
     4081 attagagaag cgttcggtgt gtttgataag gatggcaatg gctacatcag tgcagcagag
     4141 cttcgccacg tgatgacaaa ccttggagag aagttaacag atgaagaggt tgatgaaatg
     4201 atcagggaag cagacatcga tggggatggt caggtaaact acgaagagtt tgtacaaatg
     4261 atgacagcga agtgagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     4321 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     4381 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     4441 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     4501 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     4561 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     4621 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     4681 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     4741 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     4801 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     4861 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     4921 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     4981 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     5041 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5101 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     5161 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5221 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     5281 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     5341 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     5401 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     5461 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5521 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5581 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5641 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5701 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5761 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5821 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5881 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5941 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     6001 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     6061 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6121 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6181 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6241 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6301 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     6361 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     6421 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     6481 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     6541 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     6601 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     6661 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     6721 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     6781 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6841 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6901 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6961 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     7021 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     7081 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     7141 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     7201 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     7261 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     7321 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7381 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7441 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7501 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7561 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat
//
