LOCUS       sequence-198911-        6648 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Genetically encoded voltage indicator (GEVI) ASAP2s in AAV vector 
            for expression in excitatory glutamatergic neurons using the 
            promoter CaMKII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6648)
  AUTHORS   Chamberland S, Yang HH, Pan MM, Evans SW, Guan S, Chavarha M, Yang 
            Y, Salesse C, Wu H, Wu JC, Clandinin TR, Toth K, Lin MZ, St-Pierre F
  TITLE     Fast two-photon imaging of subcellular voltage dynamics in neuronal 
            tissue with genetically encoded indicators.
  JOURNAL   eLife. 2017 Jul 27;6. pii: e25690. doi: 10.7554/eLife.25690.
  PUBMED    28749338
REFERENCE   2  (bases 1 to 6648)
  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: "eLife."; 
            date: "2017-07-27"; volume: "6. pii: e25690. doi"; pages: " 
            10.7554/eLife.25690"
COMMENT     Sequence Label: pAAV-CaMKII-ASAP2s
FEATURES             Location/Qualifiers
     source          1..6648
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        143..1431
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1452..1461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2769..3357
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2822..2842)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3240..3251)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3389..3865
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3588..3607)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3905..4045
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3905..4034
                     /label=AAV2 ITR
     rep_origin      4120..4575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4207..4226)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4417..4438
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4592..4611)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4711..4733
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4771..4789)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4857..4961
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4962..5030,5031..5822)
                     /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(5180..5199)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5993..6581
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6482..6501
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6643..6648,1..124)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc
       61 ccggcctcag tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt
      121 tcctgcggcc gcacgcgttt aacattatgg ccttaggtca cttcatctcc atggggttct
      181 tcttctgatt ttctagaaaa tgagatgggg gtgcagagag cttcctcagt gacctgccca
      241 gggtcacatc agaaatgtca gagctagaac ttgaactcag attactaatc ttaaattcca
      301 tgccttgggg gcatgcaagt acgatataca gaaggagtga actcattagg gcagatgacc
      361 aatgagttta ggaaagaaga gtccagggca gggtacatct acaccacccg cccagccctg
      421 ggtgagtcca gccacgttca cctcattata gttgcctctc tccagtccta ccttgacggg
      481 aagcacaagc agaaactggg acaggagccc caggagacca aatcttcatg gtccctctgg
      541 gaggatgggt ggggagagct gtggcagagg cctcaggagg ggccctgctg ctcagtggtg
      601 acagataggg gtgagaaagc agacagagtc attccgtcag cattctgggt ctgtttggta
      661 cttcttctca cgctaaggtg gcggtgtgat atgcacaatg gctaaaaagc agggagagct
      721 ggaaagaaac aaggacagag acagaggcca agtcaaccag accaattccc agaggaagca
      781 aagaaaccat tacagagact acaaggggga agggaaggag agatgaatta gcttcccctg
      841 taaaccttag aacccagctg ttgccagggc aacggggcaa tacctgtctc ttcagaggag
      901 atgaagttgc cagggtaact acatcctgtc tttctcaagg accatcccag aatgtggcac
      961 ccactagccg ttaccatagc aactgcctct ttgccccact taatcccatc ccgtctgtta
     1021 aaagggccct atagttggag gtgggggagg taggaagagc gatgatcact tgtggactaa
     1081 gtttgttcgc atccccttct ccaaccccct cagtacatca ccctggggga acagggtcca
     1141 cttgctcctg ggcccacaca gtcctgcagt attgtgtata taaggccagg gcaaagagga
     1201 gcaggtttta aagtgaaagg caggcaggtg ttggggaggc agttaccggg gcaacgggaa
     1261 cagggcgttt cggaggtggt tgccatgggg acctggatgc tgacgaaggc tcgcgaggct
     1321 gtgagcagcc acagtgccct gctcagaagc cccaagctcg tcagtcaagc cggttctccg
     1381 tttgcactca ggagcacggg caggcgagtg gcccctagtt ctgggggcag ctctagagcg
     1441 gtaccggatc cgccaccatg gagacgactg tgaggtatga acaggggtca gagctcacta
     1501 aaacttcgag ctctccaaca gcagatgagc ccacgataaa gattgatgat ggtcgtgatg
     1561 agggtaatga acaagacagc tgttccaata ccattaggag aaaaatttcc ccgtttgtga
     1621 tgtcatttgg attcagagta tttggagttg tgcttatcat tgtagacatc atagtggtga
     1681 ttgtggatct ggccatcagt gagaagaaaa gaggcattag agagattctt gaaggtgttt
     1741 ccctggctat agcactcttc ttccttgttg atgttctcat gagagtgttt gttgaaggct
     1801 tcaagaacta tttccggtcc aaactgaata ctttggatgc agtcatagta gtgggcactc
     1861 tgctaattaa tatgacctac tccttctctg accttgctgc ctttaacagc cataacgtgt
     1921 atattaccgc ggataaacag aaaaacggca ttaaagcgaa ctttaccgtg cgccataacg
     1981 tggaagatgg cagcgtgcag ctggcggatc attatcagca gaacaccccg attggcgatg
     2041 gcccggtgct gctgccggat aaccattatc tgagcaccca gaccgtgctg agcaaagatc
     2101 cgaacgaaaa acgcgatcac atggtgctgc tggaatttgt gaccgcagcg ggcattacac
     2161 acggcatgga tgaactgtat ggcggcaccg gcggcagcgc gagccagggc gaagaactgt
     2221 ttaccggcgt ggtgccgatt ctggtggaac tggatggcga tgtgaacggc cataaattta
     2281 gcgtgcgcgg cgaaggcgaa ggcgatgcga ccattggcaa actgaccctg aaatttattt
     2341 gcaccaccgg caaactaccg gtgccgtggc cgaccctggt gaccacctta acctatggcg
     2401 tgcagtgctt tagccgctat ccggatcata tgaaacgcca tgattttttt aaaagcgcga
     2461 tgccggaagg ctatgtgcag gaacgcacca ttagctttaa agatgatggc aaatataaaa
     2521 cccgcgcggt ggtgaaattt gaaggcgata ccctggtgaa ccgcattgaa ctgaaaggca
     2581 ccgattttaa agaagatggc aacattctgg ggcataaact ggaatataac acagatcaga
     2641 tgccgcagat ggttactctt cttcgagttc tgagaattgt tatcttaata agaatatttc
     2701 agctggcttc acagaagaaa caacttgaag tggtaaccta atgagaattc gatatcaagc
     2761 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2821 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2881 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2941 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     3001 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccggaact ttcgctttcc
     3061 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     3121 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3181 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3241 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     3301 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3361 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc
     3421 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag
     3481 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt
     3541 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc
     3601 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc
     3661 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag
     3721 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa
     3781 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg
     3841 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg
     3901 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     3961 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     4021 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg
     4081 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat
     4141 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag
     4201 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     4261 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     4321 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     4381 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     4441 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg
     4501 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     4561 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     4621 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     4681 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4741 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta
     4801 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4861 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4921 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4981 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5041 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5101 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5161 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5221 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5281 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5341 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5401 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5461 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5521 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5581 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5641 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5701 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5761 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5821 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5881 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5941 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6001 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6061 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6121 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6181 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6241 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6301 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6361 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6421 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6481 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6541 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6601 ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgca
//
