LOCUS       sequence-401140-        6063 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses somatically enriched ASAP4b in neurons, can be used to 
            package AAV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6063)
  AUTHORS   Evans SW, Shi DQ, Chavarha M, Plitt MH, Taxidis J, Madruga B, Fan 
            JL, Hwang FJ, van Keulen SC, Suomivuori CM, Pang MM, Su S, Lee S, 
            Hao YA, Zhang G, Jiang D, Pradhan L, Roth RH, Liu Y, Dorian CC, 
            Reese AL, Negrean A, Losonczy A, Makinson CD, Wang S, Clandinin TR, 
            Dror RO, Ding JB, Ji N, Golshani P, Giocomo LM, Bi GQ, Lin MZ
  TITLE     A positively tuned voltage indicator for extended electrical 
            recordings in the brain.
  JOURNAL   Nat Methods. 2023 Jul;20(7):1104-1113. doi: 
            10.1038/s41592-023-01913-z. Epub 2023 Jul 6.
  PUBMED    37429962
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"; doi: 
            "10.1038/s41592-023-01913-z"; journalName: "Nat Methods"; date: 
            "2023-07"; volume: "20"; issue: "7"; pages: "1104-1113"
COMMENT     Sequence Label: AAV-hSyn-ASAP4b-Kv-WPRE
FEATURES             Location/Qualifiers
     source          1..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      642..651
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2190..2778
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2243..2263)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2661..2672)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2810..3286
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3009..3028)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3326..3466
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3326..3455
                     /label=AAV2 ITR
     rep_origin      3541..3996
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3628..3647)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3838..3859
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4013..4032)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4132..4154
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4192..4210)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4278..4382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4383..4451,4452..5243)
                     /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(4601..4620)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5414..6002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5903..5922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc cgccaccatg gagacgactg
      661 tgaggtatga acaggggtca gagctcacta aaacttcgag ctctccaaca gcagatgagc
      721 ccacgataaa gattgatgat ggtcgtgatg agggtaatga acaagacagc tgttccaata
      781 ccattaggag aaaaatttcc ccgtttgtga tgtcatttgg attcagagta tttggagttg
      841 tgcttatcat tgtagacatc atagtggtga ttgtggatct ggccatcagt gagaagaaaa
      901 gaggcattag agagattctt gaaggtgttt ccctggctat agcactcttc ttccttgttg
      961 atgttctcat gagagtgttt gttgaaggct tcaagaacta tttccggtcc aaactgaata
     1021 ctttggatgc agtcatagta gtgggcactc tgctaattaa tatgacctac tccttctctg
     1081 acggtacacc cgttgatggt aacgtgtata ttaccgcgga taaacagaaa aacggcatta
     1141 aagcgaactt taccgtgcgc cataacgtgg aagatggcag cgtgcagctg gcggatcatt
     1201 atcagcagaa caccccgatt ggcgatggcc cggtgctgct gccggataac cattatctga
     1261 gccaccagac cgtgctgagc aaagatccga acgaaaaacg cgatcacatg gtgctgctgg
     1321 aatttgtgac cgcagcgggc attacacacg gcatggatga actgtatggc ggcaccggcg
     1381 gcagcgcgag ccagggcgaa gaactgttta ccggcgtggt gccgattctg gtggaactgg
     1441 atggcgatgt gaacggccat aaatttagcg tgcgcggcga aggcgaaggc gatgcgacca
     1501 ttggcaaact gaccctgaaa tttatttgca ccaccggcaa actaccggtg ccgtggccga
     1561 ccctggtgac caccttaacc tatggcgtgc agtgctttag ccgctatccg gatcatatga
     1621 aacgccatga tttttttaaa agcgcgatgc cggaaggcta tgtgcaggaa cgcaccatta
     1681 gctttaaaga tgatggcaaa tataaaaccc gcgcggtggt gaaatttgaa ggcgataccc
     1741 tggtgaaccg cattgaactg aaaggcaccg attttaaaga agatggcaac attctggggc
     1801 ataaactgga atataacaca gatcagatgc cgcagatggt tactcttctt cgagttctga
     1861 gaattgttat cttaataaga ataggtcagc tggcttcaca gaagaaacaa cttgaagtgg
     1921 taacccttaa gggttcttca ggttcatctg gatcttccac gcgtcagagc cagccgattc
     1981 tgaacaccaa agaaatggcg ccgcagagca aaccgccgga agaactggaa atgagcagca
     2041 tgccgagccc ggtggcgccg ctgccggcgc gcaccgaagg cgtgattgat atgcgcagca
     2101 tgagcagcat tgatagcttt attagctgcg cgaccgattt tccggaagcg acccgctttt
     2161 gataagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa
     2221 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2281 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2341 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2401 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2461 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2521 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2581 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga
     2641 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2701 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2761 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
     2821 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
     2881 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
     2941 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
     3001 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
     3061 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
     3121 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
     3181 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
     3241 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
     3301 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
     3361 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
     3421 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
     3481 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
     3541 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3601 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3661 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3721 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     3781 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3841 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     3901 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     3961 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     4021 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     4081 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4141 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     4201 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     4261 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4321 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4381 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4441 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4501 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4561 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4621 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4681 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4741 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4801 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4861 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4921 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4981 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5041 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5101 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5161 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5221 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5281 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5341 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5401 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5461 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5521 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     5581 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5641 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5701 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5761 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5821 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5881 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5941 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6001 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6061 tgt
//
