LOCUS       sequence-318373-        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-jGCaMP8m-WPRE
FEATURES             Location/Qualifiers
     source          1..5849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        44..491
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    521..554
                     /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    617..650
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1220..1239
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1479..1500
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1606..1627)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(1909..1926)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(1946..1979)
                     /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(2050..2083)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2125..2713
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2178..2198)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2596..2607)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2756..2877
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2767..2786)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2821..2840
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3039..3168
                     /label=AAV2 ITR
     rep_origin      complement(3405..3993)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3485..3504)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4164..4955,4956..5024))
                     /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     4787..4806
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5025..5129)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5155..5610)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5292..5313)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5504..5523
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5682..5811
                     /label=AAV2 ITR
ORIGIN
        1 tagccatgct ctaggaagat ctctgcagag ggccctgcgt atgagtgcaa gtgggtttta
       61 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      121 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      181 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      241 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      301 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      361 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      421 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      481 tgagagcgca gtcgaattca agctgctagc aaggatcccc ataacttcgt ataaagtatc
      541 ctatacgaag ttatatcaaa ataggaagac caatgcttca ccatcgaccc gaattgccaa
      601 gcatcaccat cgacccataa cttcgtataa tgtatgctat acgaagttat actagagctt
      661 tcacttcgct gtcatcattt gtacaaactc ttcgtagttt acctgaccat ccccatcgat
      721 gtctgcttcc ctgatcattt catcaacctc ttcatctgtt aacttctctc caaggtttgt
      781 catcacgtgg cgaagctctg ctgcactgat gtagccattg ccatccttat caaacacacc
      841 gaacgcttct ctaatttctt cttccgtgtc cctgtatttc atttttcttg ccatcattgt
      901 caggaactca gggaagtcga ttgtgccgtc accgtcggca tctacttcat tgatcatgtc
      961 ctgcagctct gcttctgtgg ggttctgccc cagagaccgc atcaccgtcc ccagctcctt
     1021 ggttgttatt gtcccatccc cgtccttgtc aaatagggag aaagcctctt tgtattctgc
     1081 gatctgctct tcagtcagtt ggtccggcag gttgtactcc agcttgtgcc ccaggatgtt
     1141 gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc
     1201 gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc
     1261 ctggatgtag ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg
     1321 gtagcggctg aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac
     1381 gggcagcttg ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc
     1441 ctcaccctcg ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag
     1501 gatgggcacc accccggtga acagctcctc gcccttgctc accatgctcc ctccggtacc
     1561 gcccttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag
     1621 gaccatgtga tcgcgcttct cgttggggtc tttcgaaagt ttggactcca cgctcaggta
     1681 gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggta
     1741 ggcgagctgc acgccgccgt cctcgatgtt gtggcggatg tggaagttcg ccttgatgcc
     1801 gttcttctgc ttgtcggcct tgatatagac gttgatcttc aggcccatca ggcggccgat
     1861 gatcttcacg gcggtggcca cctccttgaa ggtcttcttg cgacgcgtgt gatgatggtg
     1921 atgatgcatg gtggcggatc ctagtataac ttcgtatagg atactttata cgaagttatc
     1981 attgggattc ttcctatttt gatccaagca tcaccatcga ccctctagtc cagatctcac
     2041 catcgaccca taacttcgta tagcatacat tatacgaagt tatgtccctc gaagaggttc
     2101 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg
     2161 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct
     2221 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg
     2281 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact
     2341 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc
     2401 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc
     2461 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa
     2521 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc
     2581 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg
     2641 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg
     2701 gccgcctccc cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga catgataaga
     2761 tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt
     2821 gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa acaagttaac
     2881 aacaacaatt gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa
     2941 agcaagtaaa acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta
     3001 cgtagataag tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt
     3061 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3121 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga
     3181 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     3241 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     3301 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3361 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3421 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3481 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3541 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3601 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3661 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3721 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3781 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3841 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3901 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3961 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4021 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     4081 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     4141 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     4201 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     4261 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     4321 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     4381 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     4441 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     4501 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     4561 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     4621 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     4681 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     4741 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     4801 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     4861 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     4921 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     4981 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     5041 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     5101 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg
     5161 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     5221 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     5281 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     5341 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     5401 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     5461 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     5521 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     5581 ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt
     5641 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca
     5701 ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     5761 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa
     5821 tgattaaccc gccatgctac ttatctacg
//
