LOCUS       sequence-229527-        5443 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 5443)
  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 5443)
  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..5443
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      641..650
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(758..766,767..784,785..787,788..820,821..823,824..847,
                     848..874,875..931,932..937,938..1207,1208..1225,1226..1660,
                     1661..1666,1667..2107)
                     /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      854..863
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1144..1165
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1271..1292)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1532..1551)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2111..2131
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2256..2377
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2293..2312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2347..2366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2384..2839)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2521..2542)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2733..2752
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2866..2970
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2972..3329
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2972..3163
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2995..3015)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3180..3315
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3364..4158
                     /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             3364..4158
                     /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(3418..3437)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4028..4047
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4346..4365)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4390..4437
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4766..5354
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5255..5274
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag cgctaccgga ctcagatctc gccaccatgg gcatggacga
      661 gctgtacaag ggtaccgctg caaccaaaat tcaggctgcc ttccgtggac acataacaag
      721 gaaaaagctc aaagacgaga agaagggtgc tgaattcatg ggttctcatc atcatcatca
      781 tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga
      841 cgataaggat ctcgccacca tggtcgactc atcacgtcgt aagtggaata agacaggtca
      901 cgcagtcaga gctataggtc ggctgagctc actcgagaac gtctatatca aggccgacaa
      961 gcagaagaac ggcatcaagg cgaacttcaa gatccgccac aacatcgagg acggcggcgt
     1021 gcagctcgcc taccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     1081 cgacaaccac tacctgagcg tgcagtccaa actttcgaaa gaccccaacg agaagcgcga
     1141 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     1201 gtacaagggc ggtaccggag ggagcatggt gagcaagggc gaggagctgt tcaccggggt
     1261 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1321 cgagggtgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg
     1381 caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt
     1441 cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1501 ctacatccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1561 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1621 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ctgccggacc aactgactga
     1681 agagcagatc gcagaattta aagaggcttt ctccctattt gacaaggacg gggatgggac
     1741 aataacaacc aaggagctgg ggacggtgat gcggtctctg gggcagaacc ccacagaagc
     1801 agagctgcag gacatgatca atgaagtaga tgccgacggt gacggcacaa tcgacttccc
     1861 tgagttcctg acaatgatgg caagaaaagg gagctacagg gacacggaag aagaaattag
     1921 agaagcgttc ggtgtgtttg ataaggatgg caatggctac atcagtgcag cagagcttcg
     1981 ccacgtgatg acaaaccttg gagagaagtt aacagatgaa gaggttgatg aaatgatcag
     2041 ggaagcagac atcgatgggg atggtcaggt aaactacgaa gagtttgtac aaatgatgac
     2101 agcgaagcca ccaaagaaga agcgcaaggt gtaagcggcc gcgactctag atcataatca
     2161 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     2221 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     2281 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     2341 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     2401 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     2461 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     2521 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     2581 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     2641 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     2701 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     2761 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     2821 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     2881 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2941 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3001 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3061 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3121 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3181 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3241 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     3301 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     3361 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     3421 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     3481 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     3541 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     3601 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     3661 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     3721 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     3781 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     3841 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     3901 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     3961 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4021 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     4081 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     4141 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     4201 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     4261 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     4321 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     4381 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     4441 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     4501 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     4561 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     4621 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     4681 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     4741 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     4801 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     4861 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     4921 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     4981 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     5041 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     5101 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     5161 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     5221 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     5281 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     5341 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     5401 cttttgctca catgttcttt cctgcgttat cccctgattc tgt
//
