LOCUS       sequence-318382-        6295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV-mediated expression of ultrafast protein calcium sensor under 
            the CAG promoter, Cre-dependent expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6295)
  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 6295)
  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-CAG-FLEX-jGCaMP8f-WPRE
FEATURES             Location/Qualifiers
     source          1..6295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR
     promoter        485..762
                     /label=chicken beta-actin promoter
     primer_bind     1041..1063
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1125..1144
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    1179..1212
                     /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    1275..1308
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1878..1897
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2137..2158
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2264..2285)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(2567..2584)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(2604..2637)
                     /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(2708..2741)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2783..3371
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2836..2856)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3254..3265)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3414..3535
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3425..3444)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3479..3498
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3697..3826
                     /label=AAV2 ITR
     rep_origin      complement(4063..4651)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4143..4162)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4822..5613,5614..5682))
                     /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     5445..5464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5683..5787)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5813..6268)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5950..5971)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6162..6181
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc
       61 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt
      181 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag agtaccattg
      241 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      301 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
      361 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
      421 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
      481 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
      541 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg
      601 ggggggcgcg cgccrggsgg ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga
      661 ggtgcggcgg cagccaatca gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg
      721 cggcggcggc ggccctataa aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc
      781 cttcgccccg tgccccgctc cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc
      841 gcgttactcc cacaggtgag cgggcgggac ggcccttctc ctccgggctg taattagcgc
      901 ttggtttaat gacggcttgt ttcttttctg tggctgcgtg aaagccttga ggggctccgg
      961 gagggccctt tgtgcggggg gagcggctcg gggctgtccg cggggggacg gctgccttcg
     1021 ggggggacgg ggcagggcgg ggttcggctt ctggcgtgtg accggcggct ctagagcctc
     1081 tgctaaccat gttcatgcct tcttcttttt cctacagctc ctgggcaacg tgctggttat
     1141 tgtgctgtct catcattttg gcaaagaatt ggatccccat aacttcgtat aaagtatcct
     1201 atacgaagtt atatcaaaat aggaagacca atgcttcacc atcgacccga attgccaagc
     1261 atcaccatcg acccataact tcgtataatg tatgctatac gaagttatac tagagctttc
     1321 acttcgctgt catcatttgt acaaactctt cgtagtttac ctgaccatcc ccatcgatgt
     1381 ctgcttccct gatcatttca tcaacctctt catctgttaa cttctctcca aggtttgtca
     1441 tcacgtggcg aagctctgct gcactgatgt agccattgcc atccttatca aacacaccga
     1501 acgcttctct aatttcttct tccgtgtccc tgtatttcat ttttcttgcc atcattgtca
     1561 ggaactcagg gaagtcgatt gtgccgtcac cgtcggcatc tacttcattg atcatgtcct
     1621 gcagctctgc ttctgtgggg ttcagtccga gagaccgcat caccgtcccc agctccttgg
     1681 ttgttattgt cccatccccg tccttgtcaa atagggagaa agcctcttta aattctgcga
     1741 tctgctcttc agtcagttgg tccggcaggt tgtactccag cttgtgcccc aggatgttgc
     1801 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     1861 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     1921 ggatgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     1981 agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg
     2041 gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct
     2101 caccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     2161 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catgctccct ccggtaccgc
     2221 ccttgtacag ctcgtccatg ccgagagtga tcccggcggc ggtcacgaac tccagcagga
     2281 ccatgtgatc gcgcttctcg ttggggtctt tcgaaagttt ggactccacg ctcaggtagt
     2341 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtagg
     2401 cgagctgcac gccgccgtcc tcgatgttgt ggcggatgtg gaagttcgcc ttgatgccgt
     2461 tcttctgctt gtcggccttg atatagacgt tgatcttcag gcccatcagg cgggcgatga
     2521 tcttcacggc ggtggccacc tccttgaagg tcttcttgcg acgcgtgtga tgatggtgat
     2581 gatgcatggt ggcggatcct agtataactt cgtataggat actttatacg aagttatcat
     2641 tgggattctt cctattttga tccaagcatc accatcgacc ctctagtcca gatctcacca
     2701 tcgacccata acttcgtata gcatacatta tacgaagtta tgtccctcga agaggttcga
     2761 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2821 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2881 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2941 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3001 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3061 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3121 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3181 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3241 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3301 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3361 cgcctccccg catcgatacc gtcgacccgg gcggccgctt cgagcagaca tgataagata
     3421 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3481 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3541 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     3601 caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg
     3661 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     3721 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3781 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc attaatgaat
     3841 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     3901 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3961 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4021 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4081 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4141 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4201 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4261 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4321 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4381 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4441 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4501 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4561 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4621 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4681 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     4741 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     4801 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     4861 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     4921 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     4981 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5041 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5101 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5161 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5221 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5281 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5341 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5401 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5461 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5521 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5581 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5641 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     5701 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     5761 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     5821 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5881 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5941 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     6001 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     6061 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     6121 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     6181 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     6241 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caact
//
