LOCUS       sequence-150700-        6981 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intracellular Ca2+ store marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6981)
  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 6981)
  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: pEGFP-N3-TPC2
FEATURES             Location/Qualifiers
     source          1..6981
                     /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      2921..2930
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2927..2929,2930..2932,2933..3646)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2927..2929,2930..2932,2933..3643)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2972..2993)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3233..3252)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3580..3601
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3769..3890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3806..3825)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3860..3879
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3897..4352)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4034..4055)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4246..4265
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4379..4483
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4485..4842
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4485..4676
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4508..4528)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4693..4828
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4877..5671
                     /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             4877..5671
                     /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(4931..4950)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5541..5560
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5859..5878)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5903..5950
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6279..6867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6768..6787
                     /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 caagcttcga attcatggcg gaaccccagg cggagtcgga
      661 gcccctgctg ggcggggccc gcggcggtgg cggcgactgg ccggcggggc tgaccactta
      721 ccgcagcatc caagtcggcc ctggtgccgc ggccaggtgg gacctctgca ttgatcaggc
      781 tgtggtcttc atcgaagatg ctattcagta ccgctccatc aaccaccggg tggatgccag
      841 ctcgatgtgg ctttaccgac ggtattactc gaacgtatgc caacggactt tgagcttcac
      901 catcttcttg atcctgtttt tggcttttat cgagacccca tcctcactca ccagcacggc
      961 ggacgtgcgc taccgcgctg ccccctggga gccgccctgc ggcctgaccg agagtgtcga
     1021 ggtgctctgc ctgctggtct ttgcggccga cctctctgtg aagggttacc tgttcgggtg
     1081 ggcccatttc cagaaaaacc tttggctgct gggctacctc gtggtgctgg tggtgtctct
     1141 ggtggactgg accgtgtccc tgagtctcgt gtgtcatgag cccctgcgga tccgccggct
     1201 tctccgtccc ttcttcctgc tgcagaactc ctctatgatg aagaagacct tgaaatgcat
     1261 ccgctggtcg ctgccggaaa tggccagcgt cgggctgctg ctggccatcc acctgtgcct
     1321 cttcaccatg ttcggaatgc tgctgttcgc tggtgggaag caggatgatg ggcaggacag
     1381 ggagaggctg acctacttcc agaacctgcc tgagtctctg acttccctcc tggtgctgct
     1441 gaccacggcc aacaaccccg atgtgatgat tcctgcgtat tccaagaacc gggcctatgc
     1501 catcttcttc atagtcttca ctgtgatagg aagcctgttt ctgatgaacc tgctgacagc
     1561 catcatctac agtcagttcc ggggctacct gatgaaatct ctccagacct cgctgtttcg
     1621 gaggcggctg ggaacccggg ctgcctttga agtcctatcc tccatggtgg gggagggagg
     1681 agccttccct caggcagttg gggtgaagcc ccagaacttg ctgcaggtgc ttcagaaggt
     1741 ccagctggac agctcccaca aacaggccat gatggagaag gtgcgttcct acggcagtgt
     1801 tctgctgtca gctgaggagt ttcagaagct cttcaacgag cttgacagaa gtgtggttaa
     1861 agagcacccg ccgaggcccg agtaccagtc tccgtttctg cagagcgccc agttcctctt
     1921 cggccactac tactttgact acctggggaa cctcatcgcc ctggcaaacc tggtgtccat
     1981 ttgcgtgttc ctggtgctgg atgcagatgt gctgcctgct gagcgtgatg acttcatcct
     2041 ggggattctc aactgcgtct tcattgtgta ctacctgttg gagatgctgc tcaaggtctt
     2101 tgccctgggc ctgcgagggt acctgtccta ccccagcaac gtgtttgacg ggctcctcac
     2161 cgttgtcctg ctggttttgg agatctcaac tctggctgtg taccgattgc cacacccagg
     2221 ctggaggccg gagatggtgg gcctgctgtc gctgtgggac atgacccgca tgctgaacat
     2281 gctcatcgtg ttccgcttcc tgcgtatcat ccccagcatg aagccgatgg ccgtggtggc
     2341 cagtaccgtc ctgggcctgg tgcagaacat gcgtgctttt ggcgggatcc tggtggtggt
     2401 ctactacgta tttgccatca ttgggatcaa cttgtttaga ggcgtcattg tggctcttcc
     2461 tggaaacagc agcctggccc ctgccaatgg ctcggcgccc tgtgggagct tcgagcagct
     2521 ggagtactgg gccaacaact tcgatgactt tgcggctgcc ctggtcactc tgtggaactt
     2581 gatggtggtg aacaactggc aggtgtttct ggatgcatat cggcgctact caggcccgtg
     2641 gtccaagatc tattttgtat tgtggtggct ggtgtcgtct gtcatctggg tcaacctgtt
     2701 tctggccctg attctggaga acttccttca caagtgggac ccccgcagcc acctgcagcc
     2761 ccttgctggg accccagagg ccacctacca gatgactgtg gagctcctgt tcagggatat
     2821 tctggaggag cccgaggagg atgagctcac agagaggctg agccagcacc cgcacctgtg
     2881 gctgtgcagg gtcgacggta ccgcgggccc gggatccatc gccaccatgg tgagcaaggg
     2941 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     3001 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     3061 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     3121 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     3181 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     3241 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     3301 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     3361 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     3421 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     3481 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     3541 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     3601 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     3661 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3721 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3781 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3841 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3901 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3961 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4021 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4081 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4141 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4201 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4261 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4321 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     4381 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     4441 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     4501 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     4561 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     4621 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     4681 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4741 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4801 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4861 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4921 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4981 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     5041 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     5101 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     5161 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     5221 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     5281 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     5341 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     5401 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     5461 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     5521 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     5581 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     5641 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     5701 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5761 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5821 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5881 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5941 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     6001 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     6061 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     6121 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     6181 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     6241 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     6301 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     6361 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     6421 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     6481 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     6541 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     6601 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     6661 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6721 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6781 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6841 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6901 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6961 aaccgtatta ccgccatgca t
//
