LOCUS       sequence-448388-        6063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Pan-neuronal soma-targeted expression of the genetically encoded 
            voltage indicator ASAP5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  AUTHORS   Hao YA, Lee S, Roth RH, Natale S, Gomez L, Taxidis J, O'Neill PS, 
            Villette V, Bradley J, Wang Z, Jiang D, Zhang G, Sheng M, Lu D, 
            Boyden E, Delvendahl I, Golshani P, Wernig M, Feldman DE, Ji N, Ding
            J, Sudhof TC, Clandinin TR, Lin MZ
  TITLE     A fast and responsive voltage indicator with enhanced sensitivity 
            for unitary synaptic events.
  JOURNAL   Neuron. 2024 Sep 17:S0896-6273(24)00643-3. doi: 
            10.1016/j.neuron.2024.08.019.
  PUBMED    39305894
REFERENCE   2  (bases 1 to 6063)
  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: "Neuron. 
            2024 Sep 17:S0896-6273(24)00643-3. doi: 
            10.1016/j.neuron.2024.08.019."
COMMENT     Sequence Label: pAAV-hsyn-ASAP5-Kv2.1-WPRE
FEATURES             Location/Qualifiers
     source          1..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        825..1272
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1292..1301
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2840..3428
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2893..2913)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3311..3322)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3460..3936
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3659..3678)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3976..4116
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3976..4105
                     /label=AAV2 ITR
     rep_origin      4191..4646
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4278..4297)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4488..4509
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4663..4682)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4782..4804
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4842..4860)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4928..5032
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5033..5101,5102..5893)
                     /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(5251..5270)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca agtgggtttt
      841 aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      901 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      961 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
     1021 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
     1081 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
     1141 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
     1201 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
     1261 ctgagagcgc agtcgagaag gtaccggatc cgccaccatg gagacgactg tgaggtatga
     1321 acaggggtca gagctcacta aaacttcgag ctctccaaca gcagatgagc ccacgataaa
     1381 gattgatgat ggtcgtgatg agggtaatga acaagacagc tgttccaata ccattaggag
     1441 aaaaatttcc ccgtttgtga tgtcatttgg attcagagta tttggagttg tgcttatcat
     1501 tgtagacatc atagtggtga ttgtggatct ggccatcagt gagaagaaaa gaggcattag
     1561 agagattctt gaaggtgttt ccctggctat agcactcttc ttccttgttg atgttctcat
     1621 gagagtgttt gttgaaggct tcaagaacta tttccggtcc aaactgaata ctttggatgc
     1681 agtcatagta gtgggcactc tgctaattaa tatgacctac tccttctctg ctggtacatt
     1741 taaaggtgat aacgtgtata ttaccgcgga taaacagaaa aacggcatta aagcgaactt
     1801 taccgtgcgc cataacgtgg aagatggcag cgtgcagctg gcggatcatt atcagcagaa
     1861 caccccgatt ggcgatggcc cggtgctgct gccggataac cattatctga gcacccagac
     1921 cgtgctgagc aaagatccga acgaaaaacg cgatcacatg gtgctgctgg aatttgtgac
     1981 cgcagcgggc attacacacg gcatggatga actgtatggc ggcaccggcg gcagcgcgag
     2041 ccagggcgaa gaactgttta ccggcgtggt gccgattctg gtggaactgg atggcgatgt
     2101 gaacggccat aaatttagcg tgcgcggcga aggcgaaggc gatgcgacca ttggcaaact
     2161 gaccctgaaa tttatttgca ccaccggcaa actaccggtg ccgtggccga ccctggtgac
     2221 caccttaacc tatggcgtgc agtgctttag ccgctatccg gatcatatga aacgccatga
     2281 tttttttaaa agcgcgatgc cggaaggcta tgtgcaggaa cgcaccatta gctttaaaga
     2341 tgatggcaaa tataaaaccc gcgcggtggt gaaatttgaa ggcgataccc tggtgaaccg
     2401 cattgaactg aaaggcaccg attttaaaga agatggcaac attctggggc ataaactgga
     2461 atataacaca gatcagatgc cgcagatggt tactcttctt cgagttctga gaattgttat
     2521 cttaataaga gtgattcgcc tggcttcaca gaagaaacaa cttgaagtgg taacccttaa
     2581 gggttcttca ggttcatctg gatcttccac gcgtcagagc cagccgattc tgaacaccaa
     2641 agaaatggcg ccgcagagca aaccgccgga agaactggaa atgagcagca tgccgagccc
     2701 ggtggcgccg ctgccggcgc gcaccgaagg cgtgattgat atgcgcagca tgagcagcat
     2761 tgatagcttt attagctgcg cgaccgattt tccggaagcg acccgctttt gataagaatt
     2821 cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2881 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2941 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     3001 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     3061 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     3121 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     3181 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     3241 gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg
     3301 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     3361 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     3421 ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc
     3481 tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta
     3541 ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg
     3601 aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc
     3661 tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc
     3721 tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg
     3781 accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg
     3841 ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga
     3901 ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc
     3961 ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     4021 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     4081 cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc
     4141 ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg
     4201 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     4261 cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     4321 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     4381 gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg
     4441 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     4501 ccaaactgga acaacactca actctatctc gggctattct tttgatttat aagggatttt
     4561 gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     4621 taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc
     4681 cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg
     4741 tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca
     4801 gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt
     4861 tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg
     4921 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4981 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     5041 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     5101 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     5161 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     5221 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     5281 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     5341 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     5401 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     5461 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     5521 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     5581 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     5641 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5701 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5761 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5821 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5881 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5941 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     6001 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     6061 ttc
//
