LOCUS       sequence-191034-        6656 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 6656)
  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 6656)
  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..6656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        160..1448
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1469..1478
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2783..3371
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2836..2856)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3254..3265)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3403..3879
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3602..3621)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3919..4059
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3919..4048
                     /label=AAV2 ITR
     rep_origin      4134..4589
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4221..4240)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4431..4452
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4606..4625)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4725..4747
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4785..4803)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4871..4975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4976..5044,5045..5836)
                     /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(5194..5213)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6007..6595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6496..6515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtttaac attatggcct taggtcactt
      181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt
      241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt
      301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact
      361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca
      421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc
      481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat
      541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc
      601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat
      661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct
      721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc
      781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga
      841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac
      901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc
      961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa
     1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat
     1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc
     1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa
     1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt
     1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga
     1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca
     1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg
     1441 ggggcagctc tagagcggta ccggatccgc caccatggag acgactgtga ggtatgaaca
     1501 ggggtcagag ctcactaaaa cttcgagctc tccaacagca gatgagccca cgataaagat
     1561 tgatgatggt cgtgatgagg gtaatgaaca agacagctgt tccaatacca ttaggagaaa
     1621 aatttccccg tttgtgatgt catttggatt cagagtattt ggagttgtgc ttatcattgt
     1681 agacatcata gtggtgattg tggatctggc catcagtgag aagaaaagag gcattagaga
     1741 gattcttgaa ggtgtttccc tggctatagc actcttcttc cttgttgatg ttctcatgag
     1801 agtgtttgtt gaaggcttca agaactattt ccggtccaaa ctgaatactt tggatgcagt
     1861 catagtagtg ggcactctgc taattaatat gacctactcc ttctctgacc ttgctgcctt
     1921 taacagccat aacgtgtata ttaccgcgga taaacagaaa aacggcatta aagcgaactt
     1981 taccgtgcgc cataacgtgg aagatggcag cgtgcagctg gcggatcatt atcagcagaa
     2041 caccccgatt ggcgatggcc cggtgctgct gccggataac cattatctga gcacccagac
     2101 cgtgctgagc aaagatccga acgaaaaacg cgatcacatg gtgctgctgg aatttgtgac
     2161 cgcagcgggc attacacacg gcatggatga actgtatggc ggcaccggcg gcagcgcgag
     2221 ccagggcgaa gaactgttta ccggcgtggt gccgattctg gtggaactgg atggcgatgt
     2281 gaacggccat aaatttagcg tgcgcggcga aggcgaaggc gatgcgacca ttggcaaact
     2341 gaccctgaaa tttatttgca ccaccggcaa actaccggtg ccgtggccga ccctggtgac
     2401 caccttaacc tatggcgtgc agtgctttag ccgctatccg gatcatatga aacgccatga
     2461 tttttttaaa agcgcgatgc cggaaggcta tgtgcaggaa cgcaccatta gctttaaaga
     2521 tgatggcaaa tataaaaccc gcgcggtggt gaaatttgaa ggcgataccc tggtgaaccg
     2581 cattgaactg aaaggcaccg attttaaaga agatggcaac attctggggc ataaactgga
     2641 atataacaca gatcagatgc cgcagatggt tactcttctt cgagttctga gaattgttat
     2701 cttaataaga atatttcagc tggcttcaca gaagaaacaa cttgaagtgg taacctaaga
     2761 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     2821 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     2881 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     2941 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3001 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3061 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3121 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3181 atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt
     3241 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3301 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3361 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3421 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3481 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     3541 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     3601 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     3661 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     3721 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     3781 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     3841 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     3901 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     3961 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4021 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4081 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4141 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4201 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4261 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4321 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4381 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4441 gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat
     4501 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4561 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     4621 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     4681 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     4741 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     4801 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     4861 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4921 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     4981 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5041 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5101 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5161 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5221 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5281 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5341 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5401 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5461 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5521 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5581 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5641 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     5701 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5761 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5821 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     5881 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5941 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6001 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6061 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6121 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6181 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6241 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6301 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6361 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6421 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6481 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     6541 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     6601 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//
