LOCUS       sequence-241289-        5624 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 5624)
  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 5624)
  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-jGCaMP7f-WPRE
FEATURES             Location/Qualifiers
     source          1..5624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      169..178
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(175..183,184..201,202..204,205..237,238..240,241..264,
                     265..291,292..348,349..354,355..624,625..642,643..1077,
                     1078..1083,1084..1527)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKELFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYTDSEEEIREAFRVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      271..280
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     561..582
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(688..709)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(949..968)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1540..2128
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1593..1613)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2011..2022)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2131..2147)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2132..2148)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2177..2298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2188..2207)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2242..2261
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2460..2589
                     /label=AAV2 ITR
     rep_origin      complement(2826..3414)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2906..2925)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3585..4376,4377..4445))
                     /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     4208..4227
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4446..4550)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4576..5031)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4713..4734)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4925..4944
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5103..5232
                     /label=AAV2 ITR
     promoter        join(5314..5624,1..137)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc
       61 gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt
      121 cgtgcctgag agcgcagtcg aattcaagct gctagcaagg atccacccgc caccatgggt
      181 tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg
      241 gatctgtacg acgatgacga taaggatctc gccaccatgg tcgactcatc acgtcgtaag
      301 tggaataaga caggtcacgc agtcagagct ataggtcggc tgagctcact cgagaacgtc
      361 tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttcaagat ccgccacaac
      421 atcgaggacg gcggcgtgca gctcgcctac cactaccagc agaacacccc catcggcgac
      481 ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact ttcgaaagac
      541 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
      601 ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag caagggcgag
      661 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      721 aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct gaccctgaag
      781 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      841 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      901 tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga cgacggcaac
      961 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctt
     1021 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacctg
     1081 ccggaccaac tgactgaaga gcagatcgca gaatttaaag agctgttctc cctatttgac
     1141 aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg gtctctgggg
     1201 cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac
     1261 ggcacaatcg acttccctga gttcctgaca atgatggcaa gaaaaatgaa atacacagac
     1321 agtgaagaag aaattagaga agcgttccgt gtgtttgata aggatggcaa tggctacatc
     1381 agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac agatgaagag
     1441 gttgatgaaa tgatcaggga agcagacatc gatggggatg gtcaggtaaa ctacgaagag
     1501 tttgtacaaa tgatgacagc gaagtgaaag cttatcgata atcaacctct ggattacaaa
     1561 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     1621 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     1681 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     1741 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     1801 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     1861 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     1921 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt
     1981 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     2041 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     2101 cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg
     2161 cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     2221 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2281 agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg
     2341 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
     2401 gatcttccta gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa
     2461 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     2521 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     2581 agcgcgcagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     2641 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     2701 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     2761 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     2821 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     2881 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     2941 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3001 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3061 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3121 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     3181 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     3241 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     3301 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     3361 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     3421 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     3481 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     3541 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     3601 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     3661 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     3721 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     3781 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     3841 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     3901 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     3961 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     4021 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     4081 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     4141 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     4201 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     4261 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     4321 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     4381 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     4441 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     4501 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     4561 cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4621 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4681 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4741 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4801 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4861 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4921 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4981 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc
     5041 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     5101 agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct
     5161 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
     5221 ctaggggttc cttgtagtta atgattaacc cgccatgcta cttatctacg tagccatgct
     5281 ctaggaagat ctctgcagag ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat
     5341 gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca agcacccaac
     5401 ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca ggatgcggcg
     5461 aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg
     5521 cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg gtcccccgca
     5581 aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccg
//
