LOCUS       sequence-318374-        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-jGCaMP8f-WPRE
FEATURES             Location/Qualifiers
     source          1..5849
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        34..481
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    511..544
                     /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    607..640
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1210..1229
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1469..1490
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1596..1617)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(1899..1916)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(1936..1969)
                     /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(2040..2073)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2115..2703
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2168..2188)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2586..2597)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2746..2867
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2757..2776)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2811..2830
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3029..3158
                     /label=AAV2 ITR
     rep_origin      complement(3395..3983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3475..3494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4154..4945,4946..5014))
                     /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     4777..4796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5015..5119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5145..5600)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5282..5303)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5494..5513
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5672..5801
                     /label=AAV2 ITR
ORIGIN
        1 ctaggaagat ctctgcagag ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat
       61 gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca agcacccaac
      121 ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca ggatgcggcg
      181 aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg
      241 cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg gtcccccgca
      301 aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca
      361 ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga
      421 ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca
      481 gtcgaattca agctgctagc aaggatcccc ataacttcgt ataaagtatc ctatacgaag
      541 ttatatcaaa ataggaagac caatgcttca ccatcgaccc gaattgccaa gcatcaccat
      601 cgacccataa cttcgtataa tgtatgctat acgaagttat actagagctt tcacttcgct
      661 gtcatcattt gtacaaactc ttcgtagttt acctgaccat ccccatcgat gtctgcttcc
      721 ctgatcattt catcaacctc ttcatctgtt aacttctctc caaggtttgt catcacgtgg
      781 cgaagctctg ctgcactgat gtagccattg ccatccttat caaacacacc gaacgcttct
      841 ctaatttctt cttccgtgtc cctgtatttc atttttcttg ccatcattgt caggaactca
      901 gggaagtcga ttgtgccgtc accgtcggca tctacttcat tgatcatgtc ctgcagctct
      961 gcttctgtgg ggttcagtcc gagagaccgc atcaccgtcc ccagctcctt ggttgttatt
     1021 gtcccatccc cgtccttgtc aaatagggag aaagcctctt taaattctgc gatctgctct
     1081 tcagtcagtt ggtccggcag gttgtactcc agcttgtgcc ccaggatgtt gccgtcctcc
     1141 ttgaagtcga tgcccttcag ctcgatgcgg ttcaccaggg tgtcgccctc gaacttcacc
     1201 tcggcgcggg tcttgtagtt gccgtcgtcc ttgaagaaga tggtgcgctc ctggatgtag
     1261 ccttcgggca tggcggactt gaagaagtcg tgctgcttca tgtggtcggg gtagcggctg
     1321 aagcactgca cgccgtaggt cagggtggtc acgagggtgg gccagggcac gggcagcttg
     1381 ccggtggtgc agatgaactt cagggtcagc ttgccgtagg tggcatcgcc ctcaccctcg
     1441 ccggacacgc tgaacttgtg gccgtttacg tcgccgtcca gctcgaccag gatgggcacc
     1501 accccggtga acagctcctc gcccttgctc accatgctcc ctccggtacc gcccttgtac
     1561 agctcgtcca tgccgagagt gatcccggcg gcggtcacga actccagcag gaccatgtga
     1621 tcgcgcttct cgttggggtc tttcgaaagt ttggactcca cgctcaggta gtggttgtcg
     1681 ggcagcagca cggggccgtc gccgatgggg gtgttctgct ggtagtggta ggcgagctgc
     1741 acgccgccgt cctcgatgtt gtggcggatg tggaagttcg ccttgatgcc gttcttctgc
     1801 ttgtcggcct tgatatagac gttgatcttc aggcccatca ggcgggcgat gatcttcacg
     1861 gcggtggcca cctccttgaa ggtcttcttg cgacgcgtgt gatgatggtg atgatgcatg
     1921 gtggcggatc ctagtataac ttcgtatagg atactttata cgaagttatc attgggattc
     1981 ttcctatttt gatccaagca tcaccatcga ccctctagtc cagatctcac catcgaccca
     2041 taacttcgta tagcatacat tatacgaagt tatgtccctc gaagaggttc gaattcgata
     2101 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2161 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2221 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2281 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2341 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2401 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2461 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     2521 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2581 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2641 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2701 cgcatcgata ccgtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg
     2761 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     2821 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     2881 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     2941 acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag
     3001 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     3061 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3121 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcattaatga atcggccaac
     3181 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3241 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3301 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3361 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3421 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3481 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3541 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3601 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3661 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3721 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3781 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3841 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     3901 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     3961 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4021 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4081 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4141 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4201 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4261 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4321 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4381 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4441 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4501 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4561 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4621 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4681 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4741 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     4801 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4861 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4921 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4981 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5041 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5101 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa
     5161 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     5221 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     5281 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     5341 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     5401 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     5461 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     5521 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     5581 tgcgccgcta cagggcgcgt cccattcgcc attcaggctg cgcaactgtt gggaagggcg
     5641 atcggtgcgg gcctcttcgc tattacgcca gctgcgcgct cgctcgctca ctgaggccgc
     5701 ccgggcaaag cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     5761 gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa tgattaaccc
     5821 gccatgctac ttatctacgt agccatgct
//
