LOCUS       sequence-352580-        5455 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Improved genetic encoded calcium sensor based on GCaMP7b..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5455)
  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 5455)
  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-XC
FEATURES             Location/Qualifiers
     source          1..5455
                     /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"
     polyA_signal    2243..2364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2280..2299)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2334..2353
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2371..2826)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2508..2529)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2720..2739
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2853..2957
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2959..3316
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2959..3150
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2982..3002)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3167..3302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3351..4145
                     /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             3351..4145
                     /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(3405..3424)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4015..4034
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4333..4352)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4377..4424
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4753..5341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5242..5261
                     /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 agcttgctct taagttggct
     2101 ggacttgata ttggttctta agcggccgcg actctagatc ataatcagcc ataccacatt
     2161 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2221 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2281 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2341 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2401 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2461 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2521 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2581 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2641 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2701 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2761 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2821 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2881 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2941 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3001 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3061 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3121 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3181 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3241 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3301 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3361 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3421 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3481 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3541 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3601 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3661 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3721 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3781 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3841 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     3901 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3961 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     4021 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     4081 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     4141 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     4201 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     4261 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     4321 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4381 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4441 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4501 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4561 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4621 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4681 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4741 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4801 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4861 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4921 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4981 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     5041 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     5101 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     5161 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     5221 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     5281 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     5341 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5401 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
