LOCUS       sequence-401141-        6063 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses somatically enriched ASAP4e 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 Apr 14, 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-ASAP4e-Kv-WPRE
FEATURES             Location/Qualifiers
     source          1..6063
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        89..536
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      556..565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2104..2692
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2157..2177)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2575..2586)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2724..3200
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2923..2942)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3240..3380
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3240..3369
                     /label=AAV2 ITR
     rep_origin      3455..3910
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3542..3561)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3752..3773
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3927..3946)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4046..4068
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4106..4124)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4192..4296
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4297..4365,4366..5157)
                     /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(4515..4534)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5328..5916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5817..5836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5978..6063,1..44)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc gcacgcgtgt
       61 gtctagactg cagagggccc tgcgtatgag tgcaagtggg ttttaggacc aggatgaggc
      121 ggggtggggg tgcctacctg acgaccgacc ccgacccact ggacaagcac ccaaccccca
      181 ttccccaaat tgcgcatccc ctatcagaga gggggagggg aaacaggatg cggcgaggcg
      241 cgtgcgcact gccagcttca gcaccgcgga cagtgccttc gcccccgcct ggcggcgcgc
      301 gccaccgccg cctcagcact gaaggcgcgc tgacgtcact cgccggtccc ccgcaaactc
      361 cccttcccgg ccaccttggt cgcgtccgcg ccgccgccgg cccagccgga ccgcaccacg
      421 cgaggcgcga gatagggggg cacgggcgcg accatctgcg ctgcggcgcc ggcgactcag
      481 cgctgcctca gtctgcggtg ggcagcggag gagtcgtgtc gtgcctgaga gcgcagtcga
      541 gaaggtaccg gatccgccac catggagacg actgtgaggt atgaacaggg gtcagagctc
      601 actaaaactt cgagctctcc aacagcagat gagcccacga taaagattga tgatggtcgt
      661 gatgagggta atgaacaaga cagctgttcc aataccatta ggagaaaaat ttccccgttt
      721 gtgatgtcat ttggattcag agtatttgga gttgtgctta tcattgtaga catcatagtg
      781 gtgattgtgg atctggccat cagtgagaag aaaagaggca ttagagagat tcttgaaggt
      841 gtttccctgg ctatagcact cttcttcctt gttgatgttc tcatgagagt gtttgttgaa
      901 ggcttcaaga actatttccg gtccaaactg aatactttgg atgcagtcat agtagtgggc
      961 actctgctaa ttaatatgac ctactccttc tctgacggta cacccgttga tggtaacgtg
     1021 tatattaccg cggataaaca gaaaaacggc attaaagcga actttaccgt gcgccataac
     1081 gtggaagatg gcagcgtgca gctggcggat cattatcagc agaacacccc gattggcgat
     1141 ggcccggtgc tgctgccgga taaccattat ctgagccacc agaccgtgct gagcaaagat
     1201 ccgaacgaaa aacgcgatca catggtgctg ctggaatttg tgaccgcagc gggcattaca
     1261 cacggcatgg atgaactgta tggcggcacc ggcggcagcg cgagccaggg cgaagaactg
     1321 tttaccggcg tggtgccgat tctggtggaa ctggatggcg atgtgaacgg ccataaattt
     1381 agcgtgcgcg gcgaaggcga aggcgatgcg accattggca aactgaccct gaaatttatt
     1441 tgcaccaccg gcaaactacc ggtgccgtgg ccgaccctgg tgaccacctt aacctatggc
     1501 gtgcagtgct ttagccgcta tccggatcat atgaaacgcc atgatttttt taaaagcgcg
     1561 atgccggaag gctatgtgca ggaacgcacc attagcttta aagatgatgg caaatataaa
     1621 acccgcgcgg tggtgaaatt tgaaggcgat accctggtga accgcattga actgaaaggc
     1681 accgatttta aagaagatgg caacattctg gggcataaac tggaatataa cacagatcag
     1741 atgccgcaga tggttactct tcttcgagtt ctgagaattg ttatcttaat aagaataatg
     1801 cagctggctt cacagaagaa acaacttgaa gtggtaaccc ttaagggttc ttcaggttca
     1861 tctggatctt ccacgcgtca gagccagccg attctgaaca ccaaagaaat ggcgccgcag
     1921 agcaaaccgc cggaagaact ggaaatgagc agcatgccga gcccggtggc gccgctgccg
     1981 gcgcgcaccg aaggcgtgat tgatatgcgc agcatgagca gcattgatag ctttattagc
     2041 tgcgcgaccg attttccgga agcgacccgc ttttgataag aattcgatat caagcttatc
     2101 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     2161 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     2221 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     2281 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     2341 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     2401 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     2461 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     2521 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     2581 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     2641 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     2701 cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     2761 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     2821 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     2881 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     2941 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     3001 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     3061 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     3121 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     3181 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     3241 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     3301 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     3361 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     3421 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     3481 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     3541 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3601 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3661 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3721 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3781 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     3841 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3901 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     3961 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4021 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4081 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     4141 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     4201 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4261 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     4321 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     4381 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     4441 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     4501 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     4561 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     4621 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     4681 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     4741 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     4801 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     4861 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     4921 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     4981 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5041 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     5101 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     5161 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5221 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5281 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5341 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5401 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5461 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5521 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5581 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5641 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5701 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5761 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5821 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5881 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5941 cggttcctgg ccttttgctg gccttttgct cacatgtcct gcaggcagct gcgcgctcgc
     6001 tcgctcactg aggccgcccg ggcgtcgggc gacctttggt cgcccggcct cagtgagcga
     6061 gcg
//
