LOCUS       sequence-228108-        5308 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  a single-wavelength GCaMP3-based calcium indicator with improved 
            response. Please also see the GCaMP6s/m/f indicators..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5308)
  AUTHORS   Akerboom J, Chen TW, Wardill TJ, Tian L, Marvin JS, Mutlu S, 
            Calderon NC, Esposti F, Borghuis BG, Sun XR, Gordus A, Orger MB, 
            Portugues R, Engert F, Macklin JJ, Filosa A, Aggarwal A, Kerr RA, 
            Takagi R, Kracun S, Shigetomi E, Khakh BS, Baier H, Lagnado L, Wang 
            SS, Bargmann CI, Kimmel BE, Jayaraman V, Svoboda K, Kim DS, 
            Schreiter ER, Looger LL
  TITLE     Optimization of a GCaMP calcium indicator for neural activity 
            imaging.
  JOURNAL   J Neurosci. 2012 Oct 3;32(40):13819-40. doi: 
            10.1523/JNEUROSCI.2601-12.2012.
  PUBMED    23035093
REFERENCE   2  (bases 1 to 5308)
  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: 
            "10.1523/JNEUROSCI.2601-12"; journalName: "J Neurosci"; date: 
            "2012-10-3- 3"; volume: "32"; issue: "40"; pages: "13819-40"
COMMENT     Sequence Label: pCMV-GCaMP5G
FEATURES             Location/Qualifiers
     source          1..5308
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      609..618
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(615..623,624..641,642..644,645..677,678..680,681..704,
                     705..731,732..788,789..794,795..1064,1065..1082,1083..1517,
                     1518..1523,1524..1967)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(615..623,624..641,642..644,645..677,678..680,681..704,
                     705..731,732..788,789..794,795..1064,1065..1082,1083..1517,
                     1518..1523,1524..1967)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      711..720
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1001..1022
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1128..1149)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1389..1408)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2089..2210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2126..2145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2180..2199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2217..2672)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2354..2375)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2566..2585
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2699..2803
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2805..3162
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2805..2996
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2828..2848)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3013..3148
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3197..3991
                     /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             3197..3991
                     /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(3251..3270)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3861..3880
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4179..4198)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4223..4270
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4599..5187
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5088..5107
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      601 cagatctcgc caccatgggt tctcatcatc atcatcatca tggtatggct agcatgactg
      661 gtggacagca aatgggtcgg gatctgtacg acgatgacga taaggatctc gccaccatgg
      721 tcgactcatc acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc
      781 tgagctcact cgagaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga
      841 acttcaagat ccgccacaac atcgaggacg gcggcgtgca gctcgcctac cactaccagc
      901 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc
      961 agtccaaact ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     1021 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga
     1081 gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1141 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct
     1201 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     1261 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     1321 agcagcacga cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct
     1381 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1441 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1501 acaagctgga gtacaacctg ccggaccaac tgactgaaga gcagatcgca gaatttaaag
     1561 aggctttctc cctatttgac aaggacgggg atgggacaat aacaaccaag gagctgggga
     1621 cggtgatgcg gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg
     1681 aagtagatgc cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcaa
     1741 gaaaaatgaa atacacagac agtgaagaag aaattagaga agcgttccgt gtgtttgata
     1801 aggatggcaa tggctacatc agtgcagcag agcttcgcca cgtgatgaca aaccttggag
     1861 agaagttaac agatgaagag gttgatgaaa tgatcaggga agcagacatc gatggggatg
     1921 gtcaggtaaa ctacgaagag tttgtacaaa tgatgacagc gaagtgagcg gccgcgactc
     1981 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     2041 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     2101 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     2161 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     2221 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2281 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2341 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2401 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2461 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2521 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2581 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2641 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2701 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2761 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2821 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2881 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2941 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3001 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3061 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3121 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     3181 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3241 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3301 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3361 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3421 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3481 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3541 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3601 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3661 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3721 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3781 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3841 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3901 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3961 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4021 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4081 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4141 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     4201 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4261 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4321 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4381 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4441 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4501 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4561 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4621 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4681 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4741 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4801 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4861 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4921 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4981 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5041 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5101 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5161 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5221 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     5281 aaccgtatta ccgccatgca ttagttat
//
