LOCUS       sequence-241275-        5602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ultrasensitive protein calcium sensor 
            under the Syn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5602)
  AUTHORS   Dana H, Sun Y, Mohar B, Hulse BK, Kerlin AM, Hasseman JP, Tsegaye G,
            Tsang A, Wong A, Patel R, Macklin JJ, Chen Y, Konnerth A, Jayaraman 
            V, Looger LL, Schreiter ER, Svoboda K, Kim DS
  TITLE     High-performance calcium sensors for imaging activity in neuronal 
            populations and microcompartments.
  JOURNAL   Nat Methods. 2019 Jul;16(7):649-657. doi: 10.1038/s41592-019-0435-6.
            Epub 2019 Jun 17.
  PUBMED    31209382
REFERENCE   2  (bases 1 to 5602)
  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: 
            "10.1038/s41592-019-0435-6"; journalName: "Nat Methods"; date: 
            "2019-07"; volume: "16"; issue: "7"; pages: "649-657"
COMMENT     Sequence Label: pGP-AAV-syn-jGCaMP7s-WPRE
FEATURES             Location/Qualifiers
     source          1..5602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      52..61
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(58..66,67..84,85..87,88..120,121..123,124..147,
                     148..174,175..231,232..237,238..507,508..525,526..960,
                     961..966,967..1410)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRVIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      154..163
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     444..465
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(571..592)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(832..851)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1423..2011
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1476..1496)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1894..1905)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2014..2030)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2015..2031)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2060..2181
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2071..2090)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2125..2144
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(2687..3275)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2767..2786)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3446..4237,4238..4306))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4069..4088
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4307..4411)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4437..4892)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4574..4595)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4786..4805
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4964..5093
                     /label=AAV2 ITR
     promoter        join(5175..5602,1..20)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 tgtcgtgcct gagagcgcag tcgaattcaa gctgctagca aggatccacc cgccaccatg
       61 ggttctcatc atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt
      121 cgggatctgt acgacgatga cgataaggat ctcgccacca tggtcgactc atcacgtcgt
      181 aagtggaata agacaggtca cgcagtcaga gtgataggtc ggctgagctc actcgagaac
      241 gtctatatca aggccgacaa gcagaagaac ggcatcaagg cgaacttcca catccgccac
      301 aacatcgagg acggcggcgt gcagctcgcc taccactacc agcagaacac ccccatcggc
      361 gacggccccg tgctgctgcc cgacaaccac tacctgagcg tgcagtccaa actttcgaaa
      421 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
      481 actctcggca tggacgagct gtacaagggc ggtaccggag ggagcatggt gagcaagggc
      541 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
      601 cacaagttca gcgtgtccgg cgagggtgag ggcgatgcca cctacggcaa gctgaccctg
      661 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
      721 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
      781 aagtccgcca tgcccgaagg ctacatccag gagcgcacca tcttcttcaa ggacgacggc
      841 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
      901 cttaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
      961 cttcctgacc aactgactga agagcagatc gcagaattta aagagctttt ctccctattt
     1021 gacaaggacg gggatgggac aataacaacc aaggagctgg ggacggtgat gcggtctctg
     1081 gggcagaacc ccacagaagc agagctgcag gacatgatca atgaagtaga tgccgacggt
     1141 gacggcacaa tcgacttccc tgagttcctg acaatgatgg caagaaaaat gaaatacagg
     1201 gacacggaag aagaaattag agaagcgttc ggtgtgtttg ataaggatgg caatggctac
     1261 atcagtgcag cagagcttcg ccacgtgatg acaaaccttg gagagaagtt aacagatgaa
     1321 gaggttgatg aaatgatcag ggaagcagac atcgatgggg atggtcaggt aaactacgaa
     1381 gagtttgtac aaatgatgac agcgaagtga aagcttatcg ataatcaacc tctggattac
     1441 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     1501 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     1561 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     1621 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     1681 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     1741 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     1801 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     1861 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     1921 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     1981 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacctcg acccgggcgg
     2041 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     2101 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     2161 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     2221 ggggagatgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     2281 aaggatcttc ctagagcatg gctacgtaga taagtagcat ggcgggttaa tcattaacta
     2341 caaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     2401 ggccgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcattaat
     2461 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     2521 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2581 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2641 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2701 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2761 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2821 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2881 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2941 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3001 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3061 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     3121 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     3181 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     3241 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     3301 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     3361 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     3421 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     3481 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     3541 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     3601 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     3661 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     3721 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     3781 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     3841 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     3901 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     3961 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4021 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4081 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     4141 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     4201 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     4261 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     4321 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     4381 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     4441 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     4501 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     4561 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     4621 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     4681 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     4741 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     4801 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     4861 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     4921 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctgcgcg ctcgctcgct
     4981 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     5041 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt ccttgtagtt
     5101 aatgattaac ccgccatgct acttatctac gtagccatgc tctaggaaga tctctgcaga
     5161 gggccctgcg tatgagtgca agtgggtttt aggaccagga tgaggcgggg tgggggtgcc
     5221 tacctgacga ccgaccccga cccactggac aagcacccaa cccccattcc ccaaattgcg
     5281 catcccctat cagagagggg gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca
     5341 gcttcagcac cgcggacagt gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc
     5401 agcactgaag gcgcgctgac gtcactcgcc ggtcccccgc aaactcccct tcccggccac
     5461 cttggtcgcg tccgcgccgc cgccggccca gccggaccgc accacgcgag gcgcgagata
     5521 ggggggcacg ggcgcgacca tctgcgctgc ggcgccggcg actcagcgct gcctcagtct
     5581 gcggtgggca gcggaggagt cg
//
