LOCUS       sequence-270807-        6385 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV plasmid for CaMKII-driven expression of a genetically-encoded 
            voltage indicator nirButterfly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6385)
  AUTHORS   Monakhov MV, Matlashov ME, Colavita M, Song C, Shcherbakova DM, 
            Antic SD, Verkhusha VV, Knopfel T
  TITLE     Screening and Cellular Characterization of Genetically Encoded 
            Voltage Indicators Based on Near-Infrared Fluorescent Proteins.
  JOURNAL   ACS Chem Neurosci. 2020 Nov 4;11(21):3523-3531. doi: 
            10.1021/acschemneuro.0c00046. Epub 2020 Oct 16.
  PUBMED    33063984
REFERENCE   2  (bases 1 to 6385)
  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: 
            "10.1021/acschemneuro.0c00046"; journalName: "ACS Chem Neurosci"; 
            date: "2020-11-4- 4"; volume: "11"; issue: "21"; pages: "3523-3531"
COMMENT     Sequence Label: pAAV-CaMKII-nirButterfly
FEATURES             Location/Qualifiers
     source          1..6385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      487..496
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             808..1755
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 720 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP720
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGSVIAEDRCAEVESKLGLHYPASFIPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPNHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACKRVAERLATQIGVMEE"
     primer_bind     complement(3214..3234)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   3594..3734
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3594..3723
                     /label=AAV2 ITR
     rep_origin      3809..4264
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3896..3915)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4106..4127
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4281..4300)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4400..4422
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4460..4478)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4546..4650
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4651..4719,4720..5511)
                     /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(4869..4888)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5682..6270
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6171..6190
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6332..6385,1..76)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
       61 atcactaggg gttcctgcgg ccgcagatgc tgcacacttg tggactaagt ttgttcgcat
      121 ccccttctcc aaccccctca gtacatcacc ctgggggaac agggtccact tgctcctggg
      181 cccacacagt cctgcagtat tgtgtatata aggccagggc aaagaggagc aggttttaaa
      241 gtgaaaggca ggcaggtgtt ggggaggcag ttaccggggc aacgggaaca gggcgtttcg
      301 gaggtggttg ccatggggac ctggatgctg acgaaggctc gcgaggctgt gagcagccac
      361 agtgccctgc tcagaagccc caagctcgtc agtcaagccg gttctccgtt tgcactcagg
      421 agcacgggca ggcgagtggc ccctagttct gggggcagcg tgaaaagacg gtatcgataa
      481 gcttgagcca ccatggaagg ttttgacggg tcagacttta gccctcctgc cgatctcgtg
      541 ggtgtggacg gtgccgtgat gcggaacgtg gtggacgtga caatcaacgg cgacgtgact
      601 gcaccaccta aggcagctcc tagaaaatct gagagtgtga agaaagtcca ctggaacgac
      661 gtggatcagg gaccaagcga gaagcccgaa acaagacagg agaagagcag gatcaccagc
      721 gagggcgagt acatccccct ggaccagatc gacatcaacg tggacgacgg gaggatggaa
      781 attcctacca caggagtggg tgctagcatg gcggaaggat ccgtcgccag gcagcctgac
      841 ctcttgacct gcgacgatga gccgatccat atccccggtg ccatccaacc gcatggactg
      901 ctgctcgccc tcgccgccga catgacgatc gttgccggca gcgacaacct tcccgaactc
      961 accggactgg cgatcggcgc cctgatcggc cgctctgcgg ccgatgtctt cgactcggag
     1021 acgcacaacc gtctgacgat cgccttggcc gagcccgggg cggccgtcgg agcaccgatc
     1081 actgtcggct tcacgatgcg aaaggacgca ggcttcatcg gctcctggca tcgccatgat
     1141 cagctcatct tcctggagct ggagcctccc cagcgggacg tcgccgagcc gcaggcgttc
     1201 ttccgccgca ccaacagcgc catccgccgc ctgcaggccg ccgaaacctt ggaaagcgcc
     1261 tgcgccgccg cggcgcaaga ggtgcggaag attaccggct tcgatcgggt gatgatctat
     1321 cgcttcgcct ccgacttcag cgggtccgtg atcgcagagg atcggtgcgc cgaggtcgag
     1381 tcaaaactag gcctgcacta tcctgcctca ttcatcccgg cgcaggcccg tcggctctat
     1441 accatcaacc cggtacggat cattcccgac atcaattatc ggccggtgcc ggtcacccca
     1501 gacctcaatc cggtcaccgg gcggccgatt gatcttagct tcgccatcct gcgcagcgtc
     1561 tcgcccaacc atctggagtt catgcgcaac ataggcatgc acggcacgat gtcgatctcg
     1621 attttgcgcg gcgagcgact gtggggattg atcgtttgcc atcaccgaac gccgtactac
     1681 gtcgatctcg atggccgcca agcctgcaag agggtcgccg agaggctggc cactcagatc
     1741 ggcgtgatgg aagaggccgt agcccgcgtc cagtttcgtg tccgagcagt gattgatcat
     1801 ctagggatgc gagtctttgg agtcttccta attttcttgg acatcatcct catgatcatt
     1861 gatctcagtc ttccaggaaa aagtgaatct tcacaatcct tttatgacgg gatggctttg
     1921 gctctttctt gttatttcat gctggattta ggattaagga tatttgccta cgggcccaag
     1981 aatttcttca ccaacccctg ggaggttgct gatggtttga ttatcgttgt cacattcgtc
     2041 gtcacgatat tttacactgt gttagatgaa tacgtgcaag aaacaggagc cgatggtttg
     2101 gggcagttgg ttgtgttggc ccgtttgctg cgtatcttca agctgacccg ccacttcgtg
     2161 gcccaccaac aaatgaaggc tgaagccgcg gccaaagaag ccgcggcaaa ggaggcagca
     2221 gcgaaagaag ccgcggccaa agcccattcg aattgcgaac atgaagagat ccacctcgcc
     2281 ggctcgatcc agccgcatgg cgcgcttctg gtcgtcagcg aacatgatca tcgcgtcatc
     2341 caggccagcg ccaacgccgc ggaatttctg aatctcggaa gcgtactcgg cgttccgctc
     2401 gccgagatcg acggcgatct gttgatcaag atcctgccgc atctcgatcc caccgccgaa
     2461 ggcatgccgg tcgcggtgcg ctgccggatc ggcaatccct ctacggagta ctgcggtctg
     2521 atgcatcggc ctccggaagg cgggctgatc atcgaactcg aacgtgccgg cccgtcgatc
     2581 gatctgtcag gcacgctggc gccggcgctg gagcggatcc gcacggcggg ttcactgcgc
     2641 gcgctgtgcg atgacaccgt gctgctgttt cagcagtgca ccggctacga ccgggtgatg
     2701 gtgtatcgtt tcgatgagca aggccacggc ctggtattct ccgagtgcca tgtgcctggg
     2761 ctcgaatcct atttcggcaa ccgctatccg tcgtcgactg tcccgcagat ggcgcggcag
     2821 ctgtacgtgc ggcagcgcgt ccgcgtgctg gtcgacgtca cctatcagcc ggtgccgctg
     2881 gagccgcggc tgtcgccgct gaccgggcgc gatctcgaca tgtcgggctg cttcctgcgc
     2941 tcgatgtcgc cgtgccatct gcagttcctg aaggacatgg gcgtgcgcgc caccctggcg
     3001 gtgtcgctgg tggtcggcgg caagctgtgg ggcctggttg tctgtcacca ttatctgccg
     3061 cgcttcatcc gtttcgagct gcgggcgatc tgcaaacggc tcgccgaaag gatcgcgacg
     3121 cggatcaccg cgcttgagag ctaagaattc cgctcgagat aatcaacctc tggattacaa
     3181 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3241 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3301 cttgtataaa tcctggttag ttcttgccac ggcggaactc atcgccgcct gccttgcccg
     3361 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttta tttgtgaaat
     3421 ttgtgatgct attgctttat ttgtaaccat ctagctttat ttgtgaaatt tgtgatgcta
     3481 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     3541 attttatgtt tcaggttcag ggggagatgt gggaggtttt ttaaagcggc cgcaggaacc
     3601 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3661 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3721 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     3781 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     3841 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     3901 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     3961 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4021 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4081 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4141 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4201 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4261 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4321 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4381 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4441 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4501 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4561 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4621 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4681 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4741 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4801 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4861 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4921 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4981 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5041 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5101 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5161 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5221 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5281 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5341 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5401 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5461 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5521 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5581 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5641 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5701 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5761 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5821 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5881 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5941 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6001 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6061 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6121 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6181 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6241 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6301 ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc
     6361 actgaggccg cccgggcgtc gggcg
//
