LOCUS       sequence-204876-        6485 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of ultrasensitive protein calcium sensor from the 
            CamKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6485)
  TITLE     AAV plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6485)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AAV.CamKII.GCaMP6s.WPRE.SV40
FEATURES             Location/Qualifiers
     source          1..6485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        160..1448
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1473..1482
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1479..1487,1488..1505,1506..1508,1509..1541,1542..1544,
                     1545..1568,1569..1595,1596..1652,1653..1658,1659..1928,
                     1929..1946,1947..2381,2382..2387,2388..2831)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6s
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFHIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEAFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1575..1584
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1865..1886
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1992..2013)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2253..2272)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2844..3432
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2897..2917)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3315..3326)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3475..3596
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3486..3505)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3540..3559
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3748..3888
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3748..3877
                     /label=AAV2 ITR
     rep_origin      3963..4418
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4050..4069)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4260..4281
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4435..4454)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4554..4576
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4614..4632)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4700..4804
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4805..4873,4874..5665)
                     /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     complement(5023..5042)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5836..6424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6325..6344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct taggtcactt
      181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
      241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
      301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
      361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
      421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
      481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
      541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
      601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
      661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
      721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
      781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
      841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
      901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
      961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
     1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
     1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
     1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
     1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
     1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
     1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
     1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
     1441 ggggcagctc tagagcggta ccggatccac ccgccaccat gggttctcat catcatcatc
     1501 atcatggtat ggctagcatg actggtggac agcaaatggg tcgggatctg tacgacgatg
     1561 acgataagga tctcgccacc atggtcgact catcacgtcg taagtggaat aagacaggtc
     1621 acgcagtcag agctataggt cggctgagct cactcgagaa cgtctatatc aaggccgaca
     1681 agcagaagaa cggcatcaag gcgaacttcc acatccgcca caacatcgag gacggcggcg
     1741 tgcagctcgc ctaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1801 ccgacaacca ctacctgagc gtgcagtcca aactttcgaa agaccccaac gagaagcgcg
     1861 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1921 tgtacaaggg cggtaccgga gggagcatgg tgagcaaggg cgaggagctg ttcaccgggg
     1981 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     2041 gcgagggtga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     2101 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     2161 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     2221 gctacatcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     2281 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     2341 aggaggacgg caacatcctg gggcacaagc tggagtacaa cctgccggac caactgactg
     2401 aagagcagat cgcagaattt aaagaggctt tctccctatt tgacaaggac ggggatggga
     2461 caataacaac caaggagctg gggacggtga tgcggtctct ggggcagaac cccacagaag
     2521 cagagctgca ggacatgatc aatgaagtag atgccgacgg tgacggcaca atcgacttcc
     2581 ctgagttcct gacaatgatg gcaagaaaaa tgaaatacag ggacacggaa gaagaaatta
     2641 gagaagcgtt cggtgtgttt gataaggatg gcaatggcta catcagtgca gcagagcttc
     2701 gccacgtgat gacaaacctt ggagagaagt taacagatga agaggttgat gaaatgatca
     2761 gggaagcaga catcgatggg gatggtcagg taaactacga agagtttgta caaatgatga
     2821 cagcgaagtg aaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2881 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2941 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     3001 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     3061 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     3121 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     3181 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     3241 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     3301 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3361 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3421 ccgcctcccc gcatcgatac cgtcgacccg ggcggccgct tcgagcagac atgataagat
     3481 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3541 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     3601 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     3661 gcaagtaaaa cctctacaaa tgtggtaaaa tcgataagga tcttcctaga gcatggctac
     3721 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     3781 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     3841 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     3901 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     3961 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4021 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4081 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4141 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4201 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4261 tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt cttttgattt
     4321 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     4381 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4441 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4501 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4561 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     4621 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     4681 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     4741 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     4801 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     4861 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     4921 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     4981 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5041 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5101 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5161 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5221 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5281 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5341 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5401 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5461 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5521 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     5581 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     5641 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     5701 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     5761 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     5821 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5881 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5941 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     6001 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6061 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6121 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6181 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6241 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6301 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6361 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6421 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6481 catgt
//
