LOCUS       sequence-322411-        6249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Synaptically targeted GCaMP5G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6249)
  AUTHORS   Al Awabdh S, Gupta-Agarwal S, Sheehan DF, Muir J, Norkett R, 
            Twelvetrees AE, Griffin LD, Kittler JT
  TITLE     Neuronal activity mediated regulation of glutamate transporter GLT-1
            surface diffusion in rat astrocytes in dissociated and slice 
            cultures.
  JOURNAL   Glia. 2016 Jul;64(7):1252-64. doi: 10.1002/glia.22997.
  PUBMED    27189737
REFERENCE   2  (bases 1 to 6249)
  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: "10.1002/glia"; 
            journalName: "Glia"; date: "2016-07"; volume: "64"; issue: "7"; 
            pages: "1252-64"
COMMENT     Sequence Label: SyGCaMP5G
FEATURES             Location/Qualifiers
     source          1..6249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        52..355
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        356..559
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     510..530
                     /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(1551..1562,1563..1580,1581..1583,1584..1616,1617..1619,
                     1620..1643,1644..1670,1671..1727,1728..1733,1734..2003,
                     2004..2021,2022..2456,2457..2462,2463..2906)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWN
                     KTGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDG
                     PVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGE
                     ELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTL
                     TYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRI
                     ELKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMR
                     SLGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDG
                     NGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     CDS             join(1551..1562,1563..1580,1581..1583,1584..1616,1617..1619,
                     1620..1643,1644..1670,1671..1727,1728..1733,1734..2003,
                     2004..2021,2022..2456,2457..2462,2463..2906)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWN
                     KTGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDG
                     PVLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGE
                     ELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTL
                     TYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRI
                     ELKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMR
                     SLGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDG
                     NGYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1650..1659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1940..1961
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2067..2088)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2328..2347)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    3028..3149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3065..3084)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3119..3138
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3156..3611)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3293..3314)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3505..3524
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3638..3742
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3744..4101
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3744..3935
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3767..3787)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3952..4087
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4136..4930
                     /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             4136..4930
                     /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(4190..4209)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4800..4819
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5118..5137)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5162..5209
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5538..6126
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6027..6046
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
       61 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      121 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      181 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      241 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      301 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      361 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      421 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      481 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      541 ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca
      601 gatctcgagc caccatggac gtggtgaatc agctggtggc tgggggtcag ttccgggtgg
      661 tcaaggagcc ccttggcttc gtgaaggtgc tgcagtgggt ctttgccatc ttcgcctttg
      721 ctacgtgtgg cagctacacc ggggagcttc ggctgagcgt ggagtgtgcc aacaagacgg
      781 agagtgccct caacatcgaa gttgaattcg agtacccctt caggctgcac caagtgtact
      841 ttgatgcacc ctcctgcgtc aaagggggca ctaccaagat cttcctggtt ggggactact
      901 cctcgtcggc tgaattcttt gtcaccgtgg ctgtgtttgc cttcctctac tccatggggg
      961 ccctggccac ctacatcttc ctgcagaaca agtaccgaga gaacaacaaa gggcctatga
     1021 tggactttct ggctacagcc gtgttcgctt tcatgtggct agttagttca tcagcctggg
     1081 ccaaaggcct gtccgatgtg aagatggcca cggacccaga gaacattatc aaggagatgc
     1141 ccatgtgccg ccagacaggg aacacatgca aggaactgag ggaccctgtg acttcaggac
     1201 tcaacaccac agtggtgttt ggcttcctga acctggtgct ctgggttggc aacttatggt
     1261 tcgtgttcaa ggagacaggc tgggcagccc cattcatgcg cgcacctcca ggcgccccgg
     1321 aaaagcaacc agcacctggc gatgcctacg gcgatgcggg ctacgggcag ggccccggag
     1381 gctatgggcc ccaggactcc tacgggcctc agggtggtta tcaacccgat tacgggcagc
     1441 cagccagcgg tggcggtggc tacgggcctc agggcgacta tgggcagcaa ggctatggcc
     1501 aacagggtgc gcccacctcc ttctccaatc aggatccacc ggtcgccacc atgcggggtt
     1561 ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa atgggtcggg
     1621 atctgtacga cgatgacgat aaggatctcg ccaccatggt cgactcatca cgtcgtaagt
     1681 ggaataagac aggtcacgca gtcagagcta taggtcggct gagctcactc gagaacgtct
     1741 atatcaaggc cgacaagcag aagaacggca tcaaggcgaa cttcaagatc cgccacaaca
     1801 tcgaggacgg cggcgtgcag ctcgcctacc actaccagca gaacaccccc atcggcgacg
     1861 gccccgtgct gctgcccgac aaccactacc tgagcgtgca gtccaaactt tcgaaagacc
     1921 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     1981 tcggcatgga cgagctgtac aagggcggta ccggagggag catggtgagc aagggcgagg
     2041 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
     2101 agttcagcgt gtccggcgag ggtgagggcg atgccaccta cggcaagctg accctgaagt
     2161 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct
     2221 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
     2281 ccgccatgcc cgaaggctac atccaggagc gcaccatctt cttcaaggac gacggcaact
     2341 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
     2401 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaacctgc
     2461 cggaccaact gactgaagag cagatcgcag aatttaaaga ggctttctcc ctatttgaca
     2521 aggacgggga tgggacaata acaaccaagg agctggggac ggtgatgcgg tctctggggc
     2581 agaaccccac agaagcagag ctgcaggaca tgatcaatga agtagatgcc gacggtgacg
     2641 gcacaatcga cttccctgag ttcctgacaa tgatggcaag aaaaatgaaa tacacagaca
     2701 gtgaagaaga aattagagaa gcgttccgtg tgtttgataa ggatggcaat ggctacatca
     2761 gtgcagcaga gcttcgccac gtgatgacaa accttggaga gaagttaaca gatgaagagg
     2821 ttgatgaaat gatcagggaa gcagacatcg atggggatgg tcaggtaaac tacgaagagt
     2881 ttgtacaaat gatgacagcg aagtgagcgg ccgcgactct agatcataat cagccatacc
     2941 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     3001 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     3061 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     3121 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     3181 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3241 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3301 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3361 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3421 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3481 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     3541 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     3601 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     3661 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttcg
     3721 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3781 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3841 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3901 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3961 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     4021 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     4081 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     4141 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4201 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4261 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     4321 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4381 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4441 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4501 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4561 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4621 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     4681 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4741 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4801 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4861 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4921 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4981 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5041 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5101 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     5161 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     5221 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     5281 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     5341 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     5401 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     5461 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     5521 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     5581 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     5641 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     5701 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     5761 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     5821 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     5881 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     5941 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     6001 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     6061 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     6121 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     6181 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat
     6241 tagttatta
//
