LOCUS       sequence-329819-        6310 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GCaMP6f in CaMKII+ cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6310)
  AUTHORS   Keaveney MK, Rahsepar B, Tseng HA, Fernandez FR, Mount RA, Ta T, 
            White JA, Berg J, Han X
  TITLE     CaMKIIalpha-Positive Interneurons Identified via a microRNA-Based 
            Viral Gene Targeting Strategy.
  JOURNAL   J Neurosci. 2020 Dec 9;40(50):9576-9588. doi: 
            10.1523/JNEUROSCI.2570-19.2020. Epub 2020 Nov 6.
  PUBMED    33158963
REFERENCE   2  (bases 1 to 6310)
  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: 
            "10.1523/JNEUROSCI.2570-19.2020"; journalName: "J Neurosci"; date: 
            "2020-12-9- 9"; volume: "40"; issue: "50"; pages: "9576-9588"
COMMENT     Sequence Label: pAAV-CaMKII-GCaMP6f
FEATURES             Location/Qualifiers
     source          1..6310
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        197..1485
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1499..1508
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1505..1513,1514..1531,1532..1534,1535..1567,1568..1570,
                     1571..1594,1595..1621,1622..1678,1679..1684,1685..1954,
                     1955..1972,1973..2407,2408..2413,2414..2857)
                     /codon_start=1
                     /product="improved fluorescent protein-based calcium sensor
                     (Chen et al., 2013)"
                     /label=GCaMP6f
                     /translation="MGSHHHHHHGMASMTGGQQMGRDLYDDDDKDLATMVDSSRRKWNK
                     TGHAVRAIGRLSSLENVYIKADKQKNGIKANFKIRHNIEDGGVQLAYHYQQNTPIGDGP
                     VLLPDNHYLSVQSKLSKDPNEKRDHMVLLEFVTAAGITLGMDELYKGGTGGSMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNLPDQLTEEQIAEFKEEFSLFDKDGDGTITTKELGTVMRS
                     LGQNPTEAELQDMINEVDADGDGTIDFPEFLTMMARKMKYRDTEEEIREAFGVFDKDGN
                     GYISAAELRHVMTNLGEKLTDEEVDEMIREADIDGDGQVNYEEFVQMMTAK"
     regulatory      1601..1610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1891..1912
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2018..2039)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2279..2298)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2897..3485
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2950..2970)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3368..3379)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3488..3504)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3489..3505)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3534..3655
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3545..3564)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3599..3618
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4034..4622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4114..4133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4793..5584,5585..5653))
                     /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     5416..5435
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5654..5758)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5784..6239)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5921..5942)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6133..6152
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagatct gaattccatt atggccttag gtcacttcat ctccatgggg ttcttcttct
      241 gattttctag aaaatgagat gggggtgcag agagcttcct cagtgacctg cccagggtca
      301 catcagaaat gtcagagcta gaacttgaac tcagattact aatcttaaat tccatgcctt
      361 gggggcatgc aagtacgata tacagaagga gtgaactcat tagggcagat gaccaatgag
      421 tttaggaaag aagagtccag ggcagggtac atctacacca cccgcccagc cctgggtgag
      481 tccagccacg ttcacctcat tatagttgcc tctctccagt cctaccttga cgggaagcac
      541 aagcagaaac tgggacagga gccccaggag accaaatctt catggtccct ctgggaggat
      601 gggtggggag agctgtggca gaggcctcag gaggggccct gctgctcagt ggtgacagat
      661 aggggtgaga aagcagacag agtcattccg tcagcattct gggtctgttt ggtacttctt
      721 ctcacgctaa ggtggcggtg tgatatgcac aatggctaaa aagcagggag agctggaaag
      781 aaacaaggac agagacagag gccaagtcaa ccagaccaat tcccagagga agcaaagaaa
      841 ccattacaga gactacaagg gggaagggaa ggagagatga attagcttcc cctgtaaacc
      901 ttagaaccca gctgttgcca gggcaacggg gcaatacctg tctcttcaga ggagatgaag
      961 ttgccagggt aactacatcc tgtctttctc aaggaccatc ccagaatgtg gcacccacta
     1021 gccgttacca tagcaactgc ctctttgccc cacttaatcc catcccgtct gttaaaaggg
     1081 ccctatagtt ggaggtgggg gaggtaggaa gagcgatgat cacttgtgga ctaagtttgt
     1141 tcgcatcccc ttctccaacc ccctcagtac atcaccctgg gggaacaggg tccacttgct
     1201 cctgggccca cacagtcctg cagtattgtg tatataaggc cagggcaaag aggagcaggt
     1261 tttaaagtga aaggcaggca ggtgttgggg aggcagttac cggggcaacg ggaacagggc
     1321 gtttcggagg tggttgccat ggggacctgg atgctgacga aggctcgcga ggctgtgagc
     1381 agccacagtg ccctgctcag aagccccaag ctcgtcagtc aagccggttc tccgtttgca
     1441 ctcaggagca cgggcaggcg agtggcccct agttctgggg gcagcggggc gcgccgccgc
     1501 caccatgggt tctcatcatc atcatcatca tggtatggct agcatgactg gtggacagca
     1561 aatgggtcgg gatctgtacg acgatgacga taaggatctc gccaccatgg tcgactcatc
     1621 acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc tgagctcact
     1681 cgagaacgtc tatatcaagg ccgacaagca gaagaacggc atcaaggcga acttcaagat
     1741 ccgccacaac atcgaggacg gcggcgtaca gctcgcctac cactaccagc agaacacccc
     1801 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccaaact
     1861 ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     1921 cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag
     1981 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     2041 aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct
     2101 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     2161 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     2221 cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga
     2281 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     2341 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     2401 gtacaacctg ccggaccaac tgactgaaga gcagatcgca gaatttaaag aggaattctc
     2461 cctatttgac aaggacgggg atgggacaat aacaaccaag gagctgggga cggtgatgcg
     2521 gtctctgggg cagaacccca cagaagcaga gctgcaggac atgatcaatg aagtagatgc
     2581 cgacggtgac ggcacaatcg acttccctga gttcctgaca atgatggcaa gaaaaatgaa
     2641 atacagggac acggaagaag aaattagaga agcgttcggt gtgtttgata aggatggcaa
     2701 tggctacatc agtgcagcag agcttcgcca cgtgatgaca aaccttggag agaagttaac
     2761 agatgaagag gttgatgaaa tgatcaggga agcagacatc gatggggatg gtcaggtaaa
     2821 ctacgaagag tttgtacaaa tgatgacagc gaagtgaact agtgcggccg ccctgcaggc
     2881 atgcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     2941 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     3001 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     3061 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     3121 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     3181 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     3241 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     3301 ctttccttgg ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta
     3361 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     3421 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     3481 cccgcatcga taccgtcgac ctcgacccgg gcggccgctt cgagcagaca tgataagata
     3541 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3601 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3661 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     3721 caagtaaaac ctctacaaat gtggtaaaat cgataaggat cttcctagag catggctacg
     3781 tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg atggagttgg
     3841 ccactccctc tctgcgcgct cgctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3901 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3961 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4261 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4321 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4381 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4441 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4501 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4561 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4621 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4681 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     4741 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     4801 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     4861 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     4921 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     4981 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5041 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5101 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5161 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5221 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5281 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5341 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5401 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5461 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5521 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5581 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5641 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     5701 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     5761 catttccccg aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt
     5821 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     5881 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     5941 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6001 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6061 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6121 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6181 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6241 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     6301 attacgccag
//
