LOCUS       sequence-318372-        5849 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ultrafast protein calcium sensor under 
            the Syn promoter, Cre-dependent expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5849)
  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 5849)
  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-FLEX-jGCaMP8s-WPRE
FEATURES             Location/Qualifiers
     source          1..5849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        208..655
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    685..718
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    781..814
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1384..1403
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1643..1664
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1770..1791)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2073..2090)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2110..2143)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2214..2247)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2289..2877
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2342..2362)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2760..2771)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2920..3041
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2931..2950)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2985..3004
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3203..3332
                     /label=AAV2 ITR
     rep_origin      complement(3569..4157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3649..3668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4328..5119,5120..5188))
                     /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     4951..4970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5189..5293)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5319..5774)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5456..5477)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5668..5687
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   join(5846..5849,1..126)
                     /label=AAV2 ITR
ORIGIN
        1 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
       61 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
      121 gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctagga
      181 agatctctgc agagggccct gcgtatgagt gcaagtgggt tttaggacca ggatgaggcg
      241 gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc caacccccat
      301 tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc ggcgaggcgc
      361 gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg gcggcgcgcg
      421 ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc cgcaaactcc
      481 ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac cgcaccacgc
      541 gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg gcgactcagc
      601 gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag cgcagtcgaa
      661 ttcaagctgc tagcaaggat ccccataact tcgtataaag tatcctatac gaagttatat
      721 caaaatagga agaccaatgc ttcaccatcg acccgaattg ccaagcatca ccatcgaccc
      781 ataacttcgt ataatgtatg ctatacgaag ttatactaga gctttcactt cgctgtcatc
      841 atttgtacaa actcttcgta gtttacctga ccatccccat cgatgtctgc ttccctgatc
      901 atttcatcaa cctcttcatc tgttaacttc tctccaaggt ttgtcatcac gtggcgaagc
      961 tctgctgcac tgatgtagcc attgccatcc ttatcaaaca caccgaacgc ttctctaatt
     1021 tcttcttccg tgtccctgta tttcattttt cttgccatca ttgtcaggaa ctcagggaag
     1081 tcgattgtgc cgtcaccgtc ggcatctact tcattgatca tgtcctgcag ctctgcttct
     1141 gtggggttgt gtccgagaga ccgcatcacc gtccccagct ccttggttgt tattgtccca
     1201 tccccgtcct tgtcaaatag ggagaaagcc tctttgtatt ctgcgatctg ctcttcagtc
     1261 agttggtccg gcaggttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     1321 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     1381 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggat gtagccttcg
     1441 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
     1501 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     1561 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcacc ctcgccggac
     1621 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     1681 gtgaacagct cctcgccctt gctcaccatg ctccctccgg taccgccctt gtacagctcg
     1741 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     1801 ttctcgttgg ggtctttcga aagtttggac tccacgctca ggtagtggtt gtcgggcagc
     1861 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtaggcgag ctgcacgccg
     1921 ccgtcctcga tgttgtggcg gatgtggaag ttcgccttga tgccgttctt ctgcttgtcg
     1981 gccttgatat agacgttgat cttcaggccc atcagcatgg cgatgatctt cacggcggtg
     2041 gccacctcct tgaaggtctt cttgcgacgc gtgtgatgat ggtgatgatg catggtggcg
     2101 gatcctagta taacttcgta taggatactt tatacgaagt tatcattggg attcttccta
     2161 ttttgatcca agcatcacca tcgaccctct agtccagatc tcaccatcga cccataactt
     2221 cgtatagcat acattatacg aagttatgtc cctcgaagag gttcgaattc gatatcaagc
     2281 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2341 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2401 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2461 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2521 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2581 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2641 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     2701 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     2761 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     2821 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     2881 gataccgtcg acccgggcgg ccgcttcgag cagacatgat aagatacatt gatgagtttg
     2941 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     3001 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3061 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcaag taaaacctct
     3121 acaaatgtgg taaaatcgat aaggatcttc ctagagcatg gctacgtaga taagtagcat
     3181 ggcgggttaa tcattaacta caaggaaccc ctagtgatgg agttggccac tccctctctg
     3241 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3301 cgggcggcct cagtgagcga gcgagcgcgc agctgcatta atgaatcggc caacgcgcgg
     3361 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3421 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3481 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3541 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3601 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3661 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3721 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3781 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3841 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3901 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3961 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4021 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4081 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4141 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4201 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4261 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4321 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4381 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4441 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4501 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4561 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4621 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4681 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4741 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4801 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4861 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4921 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4981 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5041 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5101 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5161 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5221 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5281 taggggttcc gcgcacattt ccccgaaaag tgccacctaa attgtaagcg ttaatatttt
     5341 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     5401 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     5461 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     5521 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     5581 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     5641 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5701 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5761 gctacagggc gcgtcccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     5821 gcgggcctct tcgctattac gccagctgc
//
