LOCUS       sequence-318368-        5540 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ultrafast protein calcium sensor under 
            the Syn promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5540)
  AUTHORS   Zhang Y, Rozsa M, Bushey D, Zheng J, Reep D, Broussard GJ, Tsang A, 
            Tsegaye G, Patel R, Narayan S, Li JX, Zhang R, Ahrens MB, Turner GC,
            Wang SS-H, Svoboda K, Korff W, Schreiter ER, Hasseman JP, Kolb I, 
            Looger LL
  TITLE     jGCaMP8 Fast Genetically Encoded Calcium Indicators
  JOURNAL   Janelia Research Campus, Online Resource (2020) 
            doi:10.25378/janelia.13148243.
REFERENCE   2  (bases 1 to 5540)
  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: "Janelia 
            Research Campus, Online Resource (2020) 
            doi:10.25378/janelia.13148243."
COMMENT     Sequence Label: pGP-AAV-syn-jGCaMP8m-WPRE
FEATURES             Location/Qualifiers
     source          1..5540
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        124..571
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             612..629
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     911..932
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1038..1059)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1299..1318)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1890..2478
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1943..1963)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2361..2372)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2481..2497)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2482..2498)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    2527..2648
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2538..2557)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2592..2611
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2810..2939
                     /label=AAV2 ITR
     rep_origin      complement(3176..3764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3256..3275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3935..4726,4727..4795))
                     /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     4558..4577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4796..4900)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4926..5381)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5063..5084)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5275..5294
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5453..5540,1..42)
                     /label=AAV2 ITR
ORIGIN
        1 cgcgcagaga gggagtggcc aactccatca ctaggggttc cttgtagtta atgattaacc
       61 cgccatgcta cttatctacg tagccatgct ctaggaagat ctctgcagag ggccctgcgt
      121 atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct acctgacgac
      181 cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc atcccctatc
      241 agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc
      301 gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg
      361 cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc ttggtcgcgt
      421 ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag gggggcacgg
      481 gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg cggtgggcag
      541 cggaggagtc gtgtcgtgcc tgagagcgca gtcgaattca agctgctagc aaggatccac
      601 ccgccaccat gcatcatcac catcatcaca cgcgtcgcaa gaagaccttc aaggaggtgg
      661 ccaccgccgt gaagatcatc ggccgcctga tgggcctgaa gatcaacgtc tatatcaagg
      721 ccgacaagca gaagaacggc atcaaggcga acttccacat ccgccacaac atcgaggacg
      781 gcggcgtgca gctcgcctac cactaccagc agaacacccc catcggcgac ggccccgtgc
      841 tgctgcccga caaccactac ctgagcgtgg agtccaaact ttcgaaagac cccaacgaga
      901 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
      961 acgagctgta caagggcggt accggaggga gcatggtgag caagggcgag gagctgttca
     1021 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     1081 tgtccggcga gggtgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     1141 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     1201 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     1261 ccgaaggcta catccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     1321 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     1381 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaacctg ccggaccaac
     1441 tgactgaaga gcagatcgca gaatacaaag aggctttctc cctatttgac aaggacgggg
     1501 atgggacaat aacaaccaag gagctgggga cggtgatgcg gtctctgggg cagaacccca
     1561 cagaagcaga gctgcaggac atgatcaatg aagtagatgc cgacggtgac ggcacaatcg
     1621 acttccctga gttcctgaca atgatggcaa gaaaaatgaa atacagggac acggaagaag
     1681 aaattagaga agcgttcggt gtgtttgata aggatggcaa tggctacatc agtgcagcag
     1741 agcttcgcca cgtgatgaca aaccttggag agaagttaac agatgaagag gttgatgaaa
     1801 tgatcaggga agcagacatc gatggggatg gtcaggtaaa ctacgaagag tttgtacaaa
     1861 tgatgacagc gaagtaaaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     1921 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     1981 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2041 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2101 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2161 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2221 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2281 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2341 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2401 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2461 tttgggccgc ctccccgcat cgataccgtc gacctcgacc cgggcggccg cttcgagcag
     2521 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
     2581 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
     2641 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gagatgtggg
     2701 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatcttccta
     2761 gagcatggct acgtagataa gtagcatggc gggttaatca ttaactacaa ggaaccccta
     2821 gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca
     2881 aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc
     2941 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3001 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3061 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3121 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3181 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3241 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3301 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3361 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3421 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3481 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3541 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3601 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3661 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3721 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3781 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3841 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3901 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3961 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4021 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4081 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4141 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4201 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4261 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4321 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4381 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4441 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4501 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4561 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4621 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4681 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4741 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4801 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4861 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4921 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4981 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     5041 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     5101 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     5161 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     5221 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     5281 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5341 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     5401 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctgcgcgc
     5461 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc
     5521 ggcctcagtg agcgagcgag
//
