LOCUS       sequence-315666-        6062 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid for CaMKII-driven expression of a genetically-encoded 
            calcium indicator iGECI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6062)
  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 6062)
  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
FEATURES             Location/Qualifiers
     source          1..6062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   19..148
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1947..2024
                     /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"
     primer_bind     complement(3012..3032)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     3214..3233
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3343..3483
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3343..3472
                     /label=AAV2 ITR
     rep_origin      3558..4013
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3645..3664)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3855..3876
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4030..4049)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4149..4171
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4209..4227)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4295..4399
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4400..4468,4469..5260)
                     /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(4618..4637)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5431..6019
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5920..5939
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatgtcc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
       61 gggcgtcggg cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga
      121 gtggccaact ccatcactag gggttcctgc ggccgcagat gctgcacact tgtggactaa
      181 gtttgttcgc atccccttct ccaaccccct cagtacatca ccctggggga acagggtcca
      241 cttgctcctg ggcccacaca gtcctgcagt attgtgtata taaggccagg gcaaagagga
      301 gcaggtttta aagtgaaagg caggcaggtg ttggggaggc agttaccggg gcaacgggaa
      361 cagggcgttt cggaggtggt tgccatgggg acctggatgc tgacgaaggc tcgcgaggct
      421 gtgagcagcc acagtgccct gctcagaagc cccaagctcg tcagtcaagc cggttctccg
      481 tttgcactca ggagcacggg caggcgagtg gcccctagtt ctgggggcag cgtgaaaaga
      541 cggtatcgat aagcttgatt cgagctagca ccatggcctc tcattcgaat tgcgaacatg
      601 aagagatcca cctcgccggc tcgatccagc cgcatggcgc gcttctggtc gtcagcgaac
      661 atgatcatcg cgtcatccag gccagcgcca acgccgcgga atttctgaat ctcggaagcg
      721 tactcggcgt tccgctcgcc gagatcgacg gcgatctgtt gatcaagatc ctgccgcatc
      781 tcgatcccac cgccgaaggc atgccggtcg cggtgcgctg ccggatcggc aatccctcta
      841 cggagtactg cggtctgatg catcggcctc cggaaggcgg gctgatcatc gaactcgaac
      901 gtgccggccc gtcgatcgat ctgtcaggca cgctggcgcc ggcgctggag cggatccgca
      961 cggcgggttc actgcgcgcg ctgtgcgatg acaccgtgct gctgtttcag cagtgcaccg
     1021 gctacgaccg ggtgatggtg tatcgtttcg atgagcaagg ccacggcctg gtattctccg
     1081 agtgccatgt gcctgggctc gaatcctatt tcggcaaccg ctatccgtcg tcgactgtcc
     1141 cgcagatggc gcggcagctg tacgtgcggc agcgcgtccg cgtgctggtc gacgtcacct
     1201 atcagccggt gccgctggag ccgcggctgt cgccgctgac cgggcgcgat ctcgacatgt
     1261 cgggctgctt cctgcgctcg atgtcgccgt gccatctgca gttcctgaag gacatgggcg
     1321 tgcgcgccac cctggcggtg tcgctggtgg tcggcggcaa gctgtggggc ctggttgtct
     1381 gtcaccatta tctgccgcgc ttcatccgtt tcgagctgcg ggcgatctgc aaacggctcg
     1441 ccgaaaggat cgcgacgcgg atcaccgcgc tttacaccca tgaccaactg acagaagagc
     1501 agattgcaga gttcaaagaa gccttctcat tattcgacaa ggatggggac ggcaccatca
     1561 ccacaaagga acttggcacc gttatgaggt cgcttggaca aaacccaacg gaagcagaat
     1621 tgcaggatat gatcaatgaa gtcgatgctg atggcaatgg aacgatttac tttcctgaat
     1681 ttcttactat gatggctaga aaaatgaagg acacagacag cgaagaggaa atccgagaag
     1741 cattccgtgt ttttgacaag gatgggaacg gctacatcag cgctgctgaa ttacgtcacg
     1801 tcatgacaaa cctcggggag aagttaacag atgaagaagt tgatgaaatg ataagggaag
     1861 cagatatcga tggtgatggc caagtaaact atgaagagtt tgtacaaatg atgacagcaa
     1921 aggggggggg tggcggaggt ggctccaaga ggcgctggaa gaaaaacttc attgccgtca
     1981 gcgctgccaa ccggttcaag aagatatcca gctccggggc actggtagta acctgcgacg
     2041 atgagccgat ccatatcccc ggtgccatcc aaccgcatgg actgctgctc gccctcgccg
     2101 ccgacatgac gatcgttgcc ggcagcgaca accttcccga actcaccgga ctggcgatcg
     2161 gcgccctgat cggccgctct gcggccgatg tcttcgactc ggagacgcac aaccgtctga
     2221 cgatcgcctt ggccgagccc ggggcggccg tcggagcacc gatcactgtc ggcttcacga
     2281 tgcgaaagga cgcaggcttc atcggctcct ggcatcgcca tgatcagctc atcttcctcg
     2341 agctcgagcc tccccagcgg gacgtcgccg agccgcaggc gttcttccgc cgcaccaaca
     2401 gcgccatccg ccgcctgcag gccgccgaaa ccttggaaag cgcctgcgcc gccgcggcgc
     2461 aagaggtgcg gaagattacc ggcttcgatc gggtgatgat ctatcgcttc gcctccgact
     2521 tcagcgggtc cgtgatcgca gaggatcggt gcgccgaggt cgagtcaaaa ctaggcctgc
     2581 actatcctgc ctcattcatc ccggcgcagg cccgtcggct ctataccatc aacccggtac
     2641 ggatcattcc cgatatcaat tatcggccgg tgccggtcac cccagacctc aatccggtca
     2701 ccgggcggcc gattgatctt agcttcgcca tcctgcgcag cgtctcgccc aaccatctgg
     2761 agttcatgcg caacataggc atgcacggca cgatgtcgat ctcgattttg cgcggcgagc
     2821 gactgtgggg attgatcgtt tgccatcacc gaacgccgta ctacgtcgat ctcgatggcc
     2881 gccaagcctg caagaggctc gccgagaggc tggccactca gatcggcgtg atggaagagt
     2941 gagaattccg ctcgagataa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     3001 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     3061 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt
     3121 cttgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     3181 ttgggcactg acaattccgt ggtgtttatt tgtgaaattt gtgatgctat tgctttattt
     3241 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     3301 caggttcagg gggagatgtg ggaggttttt taaagcggcc gcaggaaccc ctagtgatgg
     3361 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     3421 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     3481 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     3541 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3601 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     3661 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3721 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     3781 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3841 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     3901 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     3961 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     4021 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     4081 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     4141 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     4201 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4261 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     4321 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     4381 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     4441 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     4501 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     4561 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     4621 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     4681 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     4741 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     4801 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     4861 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     4921 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     4981 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5041 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5101 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5161 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5221 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     5281 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     5341 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     5401 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     5461 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     5521 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     5581 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     5641 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     5701 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     5761 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     5821 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     5881 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     5941 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     6001 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     6061 ct
//
