LOCUS       sequence-224897-        5442 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nuclear localized GCaMP-X sensor for monitoring calcium signal in 
            nucleus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5442)
  AUTHORS   Yang Y, Liu N, He Y, Liu Y, Ge L, Zou L, Song S, Xiong W, Liu X
  TITLE     Improved calcium sensor GCaMP-X overcomes the calcium channel 
            perturbations induced by the calmodulin in GCaMP.
  JOURNAL   Nat Commun. 2018 Apr 17;9(1):1504. doi: 10.1038/s41467-018-03719-6.
  PUBMED    29666364
REFERENCE   2  (bases 1 to 5442)
  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: "2018-04-17"; volume: "9(1):1504. doi"; pages: " 
            10.1038/s41467-018-03719-6"
COMMENT     Sequence Label: pEGFP-N1-GCaMP6m-Xn
FEATURES             Location/Qualifiers
     source          1..5442
                     /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      616..625
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(733..741,742..759,760..762,763..795,796..798,799..822,
                     823..849,850..906,907..912,913..1182,1183..1200,1201..1635,
                     1636..1641,1642..2082)
                     /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      829..838
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1119..1140
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1246..1267)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1507..1526)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2086..2106
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2230..2351
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2267..2286)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2321..2340
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2358..2813)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2495..2516)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2707..2726
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2840..2944
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2946..3303
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2946..3137
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2969..2989)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3154..3289
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3338..4132
                     /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             3338..4132
                     /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(3392..3411)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4002..4021
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4320..4339)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4364..4411
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4740..5328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5229..5248
                     /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 atctcgccac catgggcatg gacgagctgt acaagggtac cgctgcaacc
      661 aaaattcagg ctgccttccg tggacacata acaaggaaaa agctcaaaga cgagaagaag
      721 ggtgctgaat tcatgggttc tcatcatcat catcatcatg gtatggctag catgactggt
      781 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatctcgc caccatggtc
      841 gactcatcac gtcgtaagtg gaataagaca ggtcacgcag tcagagctat aggtcggctg
      901 agctcactcg agaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgaac
      961 ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag
     1021 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag
     1081 tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1141 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc
     1201 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1261 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gtgagggcga tgccacctac
     1321 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     1381 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     1441 cagcacgact tcttcaagtc cgccatgccc gaaggctaca tccaggagcg caccatcttc
     1501 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1561 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1621 aagctggagt acaacctgcc ggaccaactg actgaagagc agatcgcaga atttaaagag
     1681 gctttctccc tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg
     1741 gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa
     1801 gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat gatggcaaga
     1861 aaagggagct acagggacac ggaagaagaa attagagaag cgttcggtgt gtttgataag
     1921 gatggcaatg gctacatcag tgcagcagag cttcgccacg tgatgacaaa ccttggagag
     1981 aagttaacag atgaagaggt tgatgaaatg atcagggaag cagacatcga tggggatggt
     2041 caggtaaact acgaagagtt tgtacaaatg atgacagcga agccaccaaa gaagaagcgc
     2101 aaggtgtaag cggccgcact ctagatcata atcagccata ccacatttgt agaggtttta
     2161 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2221 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2281 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2341 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     2401 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     2461 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     2521 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     2581 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     2641 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     2701 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     2761 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     2821 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2881 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2941 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3001 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3061 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3121 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3181 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3241 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3301 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3361 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     3421 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     3481 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     3541 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     3601 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     3661 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     3721 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     3781 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     3841 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     3901 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     3961 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4021 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4081 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4141 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4201 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4261 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4321 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4381 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     4441 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     4501 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     4561 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     4621 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     4681 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     4741 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     4801 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     4861 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     4921 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     4981 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5041 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5101 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5161 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5221 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5281 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5341 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     5401 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
