LOCUS       sequence-198309-        6272 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV expression of Cre recombinase-activated ultrasensitive protein 
            calcium sensor from CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6272)
  AUTHORS   Chen TW, Wardill TJ, Sun Y, Pulver SR, Renninger SL, Baohan A, 
            Schreiter ER, Kerr RA, Orger MB, Jayaraman V, Looger LL, Svoboda K, 
            Kim DS
  TITLE     Ultrasensitive fluorescent proteins for imaging neuronal activity.
  JOURNAL   Nature. 2013 Jul 18;499(7458):295-300. doi: 10.1038/nature12354.
  PUBMED    23868258
REFERENCE   2  (bases 1 to 6272)
  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"; doi: "10"; journalName: 
            "Nature"; date: "2013-07-18- 18"; volume: "499"; issue: "7458"; 
            pages: "295-300"
COMMENT     Sequence Label: pAAV.CAG.Flex.GCaMP6m.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6272
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     367..389
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     451..470
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    505..538
                     /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    601..634
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(645..1088,1089..1094,1095..1529,1530..1547,
                     1548..1817,1818..1823,1824..1880,1881..1907,1908..1931,
                     1932..1934,1935..1967,1968..1970,1971..1988,1989..1997))
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6m
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKGSYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     primer_bind     1204..1223
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1463..1484
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1590..1611)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1892..1901
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1994..2003
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2014..2047)
                     /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(2118..2151)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2193..2781
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2246..2266)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2664..2675)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2824..2945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2835..2854)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2889..2908
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3107..3236
                     /label=AAV2 ITR
     rep_origin      complement(3473..4061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3553..3572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4232..5023,5024..5092))
                     /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     4855..4874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5093..5197)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5223..5678)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5360..5381)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5572..5591
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5750..5879
                     /label=AAV2 ITR
ORIGIN
        1 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc
       61 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc
      121 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca
      181 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg
      241 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg
      301 cggggggagc ggctcggggc tgtccgcggg gggacggctg ccttcggggg ggacggggca
      361 gggcggggtt cggcttctgg cgtgtgaccg gcggctctag agcctctgct aaccatgttc
      421 atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc
      481 attttggcaa agaattggat ccccataact tcgtataaag tatcctatac gaagttatat
      541 caaaatagga agaccaatgc ttcaccatcg acccgaattg ccaagcatca ccatcgaccc
      601 ataacttcgt ataatgtatg ctatacgaag ttatactaga gctttcactt cgctgtcatc
      661 atttgtacaa actcttcgta gtttacctga ccatccccat cgatgtctgc ttccctgatc
      721 atttcatcaa cctcttcatc tgttaacttc tctccaaggt ttgtcatcac gtggcgaagc
      781 tctgctgcac tgatgtagcc attgccatcc ttatcaaaca caccgaacgc ttctctaatt
      841 tcttcttccg tgtccctgta gctccctttt cttgccatca ttgtcaggaa ctcagggaag
      901 tcgattgtgc cgtcaccgtc ggcatctact tcattgatca tgtcctgcag ctctgcttct
      961 gtggggttct gccccagaga ccgcatcacc gtccccagct ccttggttgt tattgtccca
     1021 tccccgtcct tgtcaaatag ggagaaagcc tctttaaatt ctgcgatctg ctcttcagtc
     1081 agttggtccg gcaggttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     1141 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     1201 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggat gtagccttcg
     1261 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg gctgaagcac
     1321 tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     1381 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcacc ctcgccggac
     1441 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     1501 gtgaacagct cctcgccctt gctcaccatg ctccctccgg taccgccctt gtacagctcg
     1561 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     1621 ttctcgttgg ggtctttcga aagtttggac tgcacgctca ggtagtggtt gtcgggcagc
     1681 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtaggcgag ctgcacgccg
     1741 ccgtcctcga tgttgtggcg gatcttgaag ttcgccttga tgccgttctt ctgcttgtcg
     1801 gccttgatat agacgttctc gagtgagctc agccgaccta tagctctgac tgcgtgacct
     1861 gtcttattcc acttacgacg tgatgagtcg accatggtgg cgagatcctt atcgtcatcg
     1921 tcgtacagat cccgacccat ttgctgtcca ccagtcatgc tagccatacc atgatgatga
     1981 tgatgatgag aacccatggt ggcggatcct agtataactt cgtataggat actttatacg
     2041 aagttatcat tgggattctt cctattttga tccaagcatc accatcgacc ctctagtcca
     2101 gatctcacca tcgacccata acttcgtata gcatacatta tacgaagtta tgtccctcga
     2161 agaggttcga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     2221 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     2281 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     2341 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     2401 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     2461 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     2521 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     2581 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
     2641 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2701 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     2761 ccctttgggc cgcctccccg catcgatacc gtcgacccgg gcggccgctt cgagcagaca
     2821 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     2881 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     2941 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg
     3001 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag
     3061 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     3121 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3181 gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc
     3241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     3301 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     3361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     3421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     3481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     3541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     3601 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     3661 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     3721 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     3781 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     3841 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     3901 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     3961 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     4021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     4081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     4141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     4201 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     4261 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4321 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     4381 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4441 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4501 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     4561 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     4621 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     4681 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     4741 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     4801 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     4861 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     4921 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     4981 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5041 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     5101 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5161 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     5221 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     5281 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     5341 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     5401 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     5461 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     5521 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     5581 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     5641 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     5701 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tgcgcgctcg
     5761 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc
     5821 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt
     5881 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta ggaagagtac
     5941 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     6001 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6061 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6121 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     6181 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     6241 cacccccaat tttgtattta tttatttttt aa
//
