LOCUS       sequence-352581-        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   Geng J, Tang Y, Yu Z, Gao Y, Li W, Lu Y, Wang B, Zhou H, Li P, Liu 
            N, Wang P, Fan Y, Yang Y, Guo ZV, Liu X
  TITLE     Chronic Ca(2+) imaging of cortical neurons with long-term expression
            of GCaMP-X.
  JOURNAL   eLife. 2022 Oct 5;11:e76691. doi: 10.7554/eLife.76691.
  PUBMED    36196992
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: "eLife."; 
            date: "2022-10-5"; volume: "11:e76691. doi"; pages: " 
            10.7554/eLife.76691"
COMMENT     Sequence Label: pEGFP-N1-jGCaMP7b-XN
FEATURES             Location/Qualifiers
     source          1..5443
                     /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=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNPPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMYARKMKYRDTEEEIREAFGVFDKDGN
                     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    2231..2352
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2268..2287)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2322..2341
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2359..2814)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2496..2517)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2708..2727
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2841..2945
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2947..3304
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2947..3138
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2970..2990)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3155..3290
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3339..4133
                     /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             3339..4133
                     /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(3393..3412)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4003..4022
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4321..4340)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4365..4412
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4741..5329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5230..5249
                     /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 tcgagcttaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1621 aagctggagt acaacccacc tgaccaactg actgaagagc agatcgcaga atttaaagag
     1681 cttttctccc tatttgacaa ggacggggat gggacaataa caaccaagga gctggggacg
     1741 gtgatgcggt ctctggggca gaaccccaca gaagcagagc tgcaggacat gatcaatgaa
     1801 gtagatgccg acggtgacgg cacaatcgac ttccctgagt tcctgacaat gtacgcaaga
     1861 aaaatgaaat 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 cggccgcgac tctagatcat aatcagccat accacatttg tagaggtttt
     2161 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     2221 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     2281 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     2341 caatgtatct taaggcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     2401 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     2461 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     2521 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     2581 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     2641 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     2701 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     2761 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     2821 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2881 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2941 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3001 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3061 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3121 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3181 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3241 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     3301 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3361 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3421 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3481 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     3541 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     3601 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     3661 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     3721 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     3781 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     3841 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     3901 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3961 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4021 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4081 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4141 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4201 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4261 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     4321 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     4381 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     4441 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     4501 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     4561 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     4621 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4681 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4741 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4801 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4861 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4921 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4981 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5041 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5101 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5161 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5221 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5281 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5341 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5401 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
