LOCUS       sequence-320729-        6137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid for CaMKII-driven expression of a genetically-encoded 
            calcium indicator iGECI with a nuclear export signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6137)
  AUTHORS   Shemetov AA, Monakhov MV, Zhang Q, Canton-Josh JE, Kumar M, Chen M, 
            Matlashov ME, Li X, Yang W, Nie L, Shcherbakova DM, Kozorovitskiy Y,
            Yao J, Ji N, Verkhusha VV
  TITLE     A near-infrared genetically encoded calcium indicator for in vivo 
            imaging.
  JOURNAL   Nat Biotechnol. 2021 Mar;39(3):368-377. doi: 
            10.1038/s41587-020-0710-1. Epub 2020 Oct 26.
  PUBMED    33106681
REFERENCE   2  (bases 1 to 6137)
  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: 
            "10.1038/s41587-020-0710-1"; journalName: "Nat Biotechnol"; date: 
            "2021-03"; volume: "39"; issue: "3"; pages: "368-377"
COMMENT     Sequence Label: pAAV-CaMKII-iGECI-NES
FEATURES             Location/Qualifiers
     source          1..6137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   77..217
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   88..217
                     /label=AAV2 ITR
     primer_bind     complement(327..346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     528..548
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(624..650)
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             complement(1584..1661)
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
     regulatory      3056..3065
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     repeat_region   3487..3616
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3678..4266)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3758..3777)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4437..5228,5229..5297))
                     /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     5060..5079
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5298..5402)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5470..5488
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5526..5548)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5648..5667
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(5684..6137,1..2))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5821..5842)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6033..6052
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gtactatggt tgctttgacg tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
       61 taccgcatca ggcgcccctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
      121 gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga gcgagcgcgc
      181 agagagggag tggccaactc catcactagg ggttcctgcg gccgctttaa aaaacctccc
      241 acatctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
      301 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaacacca
      361 cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg caggcggcga
      421 tgagttccgc cgtggcaaga actaaccagg atttatacaa ggaggagaaa atgaaagcca
      481 tacgggaagc aatagcatga tacaaaggca ttaaagcagc gtatccacat agcgtaaaag
      541 gagcaacata gttaagaata ccagtcaatc tttcacaaat tttgtaatcc agaggttgat
      601 tatctcgagc ggaattctca atccagagtc aggcgttcca gaggaggcag ctgcagggtt
      661 ccacctccct cttccatcac gctgatctga gtggccagcc tctcggcgag cctcttgcag
      721 gcttggcggc catcgagatc gacgtagtac ggcgttcggt gatggcaaac gatcaatccc
      781 cacagtcgct cgccgcgcaa aatcgagatc gacatcgtgc cgtgcatgcc tatgttgcgc
      841 atgaactcca gatggttggg cgagacgctg cgcaggatgg cgaagctaag atcaatcggc
      901 cgcccggtga ccggattgag gtctggggtg accggcaccg gccgataatt gatatcggga
      961 atgatccgta ccgggttgat ggtatagagc cgacgggcct gcgccgggat gaatgaggca
     1021 ggatagtgca ggcctagttt tgactcgacc tcggcgcacc gatcctctgc gatcacggac
     1081 ccgctgaagt cggaggcgaa gcgatagatc atcacccgat cgaagccggt aatcttccgc
     1141 acctcttgcg ccgcggcggc gcaggcgctt tccaaggttt cggcggcctg caggcggcgg
     1201 atggcgctgt tggtgcggcg gaagaacgcc tgcggctcgg cgacgtcccg ctggggaggc
     1261 tcgagctcga ggaagatgag ctgatcatgg cgatgccagg agccgatgaa gcctgcgtcc
     1321 tttcgcatcg tgaagccgac agtgatcggt gctccgacgg ccgccccggg ctcggccaag
     1381 gcgatcgtca gacggttgtg cgtctccgag tcgaagacat cggccgcaga gcggccgatc
     1441 agggcgccga tcgccagtcc ggtgagttcg ggaaggttgt cgctgccggc aacgatcgtc
     1501 atgtcggcgg cgagggcgag cagcagtcca tgcggttgga tggcaccggg gatatggatc
     1561 ggctcatcgt cgcaggttac taccagtgcc ccggagctgg atatcttctt gaaccggttg
     1621 gcagcgctga cggcaatgaa gtttttcttc cagcgcctct tggagccacc tccgccaccc
     1681 cccccctttg ctgtcatcat ttgtacaaac tcttcatagt ttacttggcc atcaccatcg
     1741 atatctgctt cccttatcat ttcatcaact tcttcatctg ttaacttctc cccgaggttt
     1801 gtcatgacgt gacgtaattc agcagcgctg atgtagccgt tcccatcctt gtcaaaaaca
     1861 cggaatgctt ctcggatttc ctcttcgctg tctgtgtcct tcatttttct agccatcata
     1921 gtaagaaatt caggaaagta aatcgttcca ttgccatcag catcgacttc attgatcata
     1981 tcctgcaatt ctgcttccgt tgggttttgt ccaagcgacc tcataacggt gccaagttcc
     2041 tttgtggtga tggtgccgtc cccatccttg tcgaataatg agaaggcttc tttgaactct
     2101 gcaatctgct cttctgtcag ttggtcatgg gtgtaaagcg cggtgatccg cgtcgcgatc
     2161 ctttcggcga gccgtttgca gatcgcccgc agctcgaaac ggatgaagcg cggcagataa
     2221 tggtgacaga caaccaggcc ccacagcttg ccgccgacca ccagcgacac cgccagggtg
     2281 gcgcgcacgc ccatgtcctt caggaactgc agatggcacg gcgacatcga gcgcaggaag
     2341 cagcccgaca tgtcgagatc gcgcccggtc agcggcgaca gccgcggctc cagcggcacc
     2401 ggctgatagg tgacgtcgac cagcacgcgg acgcgctgcc gcacgtacag ctgccgcgcc
     2461 atctgcggga cagtcgacga cggatagcgg ttgccgaaat aggattcgag cccaggcaca
     2521 tggcactcgg agaataccag gccgtggcct tgctcatcga aacgatacac catcacccgg
     2581 tcgtagccgg tgcactgctg aaacagcagc acggtgtcat cgcacagcgc gcgcagtgaa
     2641 cccgccgtgc ggatccgctc cagcgccggc gccagcgtgc ctgacagatc gatcgacggg
     2701 ccggcacgtt cgagttcgat gatcagcccg ccttccggag gccgatgcat cagaccgcag
     2761 tactccgtag agggattgcc gatccggcag cgcaccgcga ccggcatgcc ttcggcggtg
     2821 ggatcgagat gcggcaggat cttgatcaac agatcgccgt cgatctcggc gagcggaacg
     2881 ccgagtacgc ttccgagatt cagaaattcc gcggcgttgg cgctggcctg gatgacgcga
     2941 tgatcatgtt cgctgacgac cagaagcgcg ccatgcggct ggatcgagcc ggcgaggtgg
     3001 atctcttcat gttcgcaggt caagaggtca ggctgcctgg cgacggagcc ttccgccatg
     3061 gtggcgctag ctcgaatcaa gcttatcgat accgtctttt cacgctgccc ccagaactag
     3121 gggccactcg cctgcccgtg ctcctgagtg caaacggaga accggcttga ctgacgagct
     3181 tggggcttct gagcagggca ctgtggctgc tcacagcctc gcgagccttc gtcagcatcc
     3241 aggtccccat ggcaaccacc tccgaaacgc cctgttcccg ttgccccggt aactgcctcc
     3301 ccaacacctg cctgcctttc actttaaaac ctgctcctct ttgccctggc cttatataca
     3361 caatactgca ggactgtgtg ggcccaggag caagtggacc ctgttccccc agggtgatgt
     3421 actgaggggg ttggagaagg ggatgcgaac aaacttagtc cacaagtgtg cagcatctgc
     3481 ggccgcagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     3541 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg
     3601 cgcagctgcc tgcaggacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     3661 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     3721 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     3781 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     3841 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     3901 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     3961 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     4021 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     4081 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     4141 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     4201 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     4261 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     4321 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     4381 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     4441 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     4501 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     4561 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     4621 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     4681 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     4741 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     4801 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     4861 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     4921 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     4981 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     5041 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     5101 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     5161 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     5221 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     5281 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     5341 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     5401 cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta
     5461 acctataaaa ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt
     5521 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     5581 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     5641 aactatgcgg catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat
     5701 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc
     5761 gaaatcggca aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt
     5821 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     5881 accgtctatc agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg
     5941 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6001 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6061 aaggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6121 gcgccgctac agggcgc
//
