LOCUS       sequence-198265-        5990 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV expression of ultrasensitive protein calcium sensor from CAG 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5990)
  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 5990)
  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.GCaMP6m.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..5990
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     375..397
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     459..478
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      511..520
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(517..525,526..543,544..546,547..579,580..582,583..606,
                     607..633,634..690,691..696,697..966,967..984,985..1419,
                     1420..1425,1426..1869)
                     /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"
     regulatory      613..622
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     903..924
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1030..1051)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1291..1310)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    1882..2470
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1935..1955)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2353..2364)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2513..2634
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2524..2543)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2578..2597
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2796..2925
                     /label=AAV2 ITR
     rep_origin      complement(3162..3750)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3242..3261)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3921..4712,4713..4781))
                     /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     4544..4563
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4782..4886)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4912..5367)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5049..5070)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5261..5280
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5439..5568
                     /label=AAV2 ITR
ORIGIN
        1 gcggcagcca atcagagcgg cgcgctccga aagtttcctt ttatggcgag gcggcggcgg
       61 cggcggccct ataaaaagcg aagcgcgcgg cgggcgggag tcgctgcgcg ctgccttcgc
      121 cccgtgcccc gctccgccgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta
      181 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt
      241 taatgacggc ttgtttcttt tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc
      301 cctttgtgcg gggggagcgg ctcggggctg tccgcggggg gacggctgcc ttcggggggg
      361 acggggcagg gcggggttcg gcttctggcg tgtgaccggc ggctctagag cctctgctaa
      421 ccatgttcat gccttcttct ttttcctaca gctcctgggc aacgtgctgg ttattgtgct
      481 gtctcatcat tttggcaaag aattggatcc gccaccatgg gttctcatca tcatcatcat
      541 catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta cgacgatgac
      601 gataaggatc tcgccaccat ggtcgactca tcacgtcgta agtggaataa gacaggtcac
      661 gcagtcagag ctataggtcg gctgagctca ctcgagaacg tctatatcaa ggccgacaag
      721 cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcggcgtg
      781 cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
      841 gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga gaagcgcgat
      901 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
      961 tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt caccggggtg
     1021 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     1081 gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     1141 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     1201 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     1261 tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     1321 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     1381 gaggacggca acatcctggg gcacaagctg gagtacaacc tgccggacca actgactgaa
     1441 gagcagatcg cagaatttaa agaggctttc tccctatttg acaaggacgg ggatgggaca
     1501 ataacaacca aggagctggg gacggtgatg cggtctctgg ggcagaaccc cacagaagca
     1561 gagctgcagg acatgatcaa tgaagtagat gccgacggtg acggcacaat cgacttccct
     1621 gagttcctga caatgatggc aagaaaaggg agctacaggg acacggaaga agaaattaga
     1681 gaagcgttcg gtgtgtttga taaggatggc aatggctaca tcagtgcagc agagcttcgc
     1741 cacgtgatga caaaccttgg agagaagtta acagatgaag aggttgatga aatgatcagg
     1801 gaagcagaca tcgatgggga tggtcaggta aactacgaag agtttgtaca aatgatgaca
     1861 gcgaagtgaa agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     1921 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     1981 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     2041 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     2101 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     2161 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     2221 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     2281 tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc
     2341 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     2401 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     2461 gcctccccgc atcgataccg tcgacccggg cggccgcttc gagcagacat gataagatac
     2521 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     2581 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     2641 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     2701 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     2761 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     2821 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     2881 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc
     2941 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3001 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3061 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3121 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3181 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3241 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3301 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3361 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3421 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3481 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3541 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3601 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3661 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3721 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3781 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3841 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3901 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3961 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4021 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4081 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4141 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4201 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4261 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4321 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4381 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4441 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4501 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4561 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4621 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4681 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4741 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4801 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4861 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa
     4921 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     4981 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     5041 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     5101 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     5161 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     5221 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     5281 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     5341 cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc aactgttggg
     5401 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct gcgcgctcgc tcgctcactg
     5461 aggccgcccg ggcaaagccc gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg
     5521 agcgagcgcg cagagaggga gtggccaact ccatcactag gggttccttg tagttaatga
     5581 ttaacccgcc atgctactta tctacgtagc catgctctag gaagagtacc attgacgtca
     5641 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
     5701 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
     5761 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
     5821 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtc
     5881 gaggtgagcc ccacgttctg cttcactctc cccatctccc ccccctcccc acccccaatt
     5941 ttgtatttat ttatttttta attattttgt gcagcgatgg gggcgggggg
//
