LOCUS       sequence-448386-        7111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre dependent 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 7111)
  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 7111)
  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-EF1a-Dio-ASAP5-Kv2.1-WPRE
FEATURES             Location/Qualifiers
     source          1..7111
                     /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        881..2059
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1112..2050
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2007..2027
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(2090..2123)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(2174..2207)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      3730..3739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3746..3779
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    3830..3863
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3888..4476
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3941..3961)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4359..4370)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4508..4984
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4707..4726)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5024..5164
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5024..5153
                     /label=AAV2 ITR
     rep_origin      5239..5694
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5326..5345)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5536..5557
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5711..5730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5830..5852
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5890..5908)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5976..6080
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6081..6149,6150..6941)
                     /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(6299..6318)
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg
      841 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
      901 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      961 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
     1021 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
     1081 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
     1141 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
     1201 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
     1261 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
     1321 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
     1381 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
     1441 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
     1501 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
     1561 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
     1621 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
     1681 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
     1741 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1801 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     1861 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1921 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1981 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     2041 tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa taacttcgta
     2101 taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc tattagcaat
     2161 atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc gcgcctcatt
     2221 aaaagcgggt cgcttccgga aaatcggtcg cgcagctaat aaagctatca atgctgctca
     2281 tgctgcgcat atcaatcacg ccttcggtgc gcgccggcag cggcgccacc gggctcggca
     2341 tgctgctcat ttccagttct tccggcggtt tgctctgcgg cgccatttct ttggtgttca
     2401 gaatcggctg gctctgacgc gtggaagatc cagatgaacc tgaagaaccc ttaagggtta
     2461 ccacttcaag ttgtttcttc tgtgaagcca ggcgaatcac tcttattaag ataacaattc
     2521 tcagaactcg aagaagagta accatctgcg gcatctgatc tgtgttatat tccagtttat
     2581 gccccagaat gttgccatct tctttaaaat cggtgccttt cagttcaatg cggttcacca
     2641 gggtatcgcc ttcaaatttc accaccgcgc gggttttata tttgccatca tctttaaagc
     2701 taatggtgcg ttcctgcaca tagccttccg gcatcgcgct tttaaaaaaa tcatggcgtt
     2761 tcatatgatc cggatagcgg ctaaagcact gcacgccata ggttaaggtg gtcaccaggg
     2821 tcggccacgg caccggtagt ttgccggtgg tgcaaataaa tttcagggtc agtttgccaa
     2881 tggtcgcatc gccttcgcct tcgccgcgca cgctaaattt atggccgttc acatcgccat
     2941 ccagttccac cagaatcggc accacgccgg taaacagttc ttcgccctgg ctcgcgctgc
     3001 cgccggtgcc gccatacagt tcatccatgc cgtgtgtaat gcccgctgcg gtcacaaatt
     3061 ccagcagcac catgtgatcg cgtttttcgt tcggatcttt gctcagcacg gtctgggtgc
     3121 tcagataatg gttatccggc agcagcaccg ggccatcgcc aatcggggtg ttctgctgat
     3181 aatgatccgc cagctgcacg ctgccatctt ccacgttatg gcgcacggta aagttcgctt
     3241 taatgccgtt tttctgttta tccgcggtaa tatacacgtt atcaccttta aatgtaccag
     3301 cagagaagga gtaggtcata ttaattagca gagtgcccac tactatgact gcatccaaag
     3361 tattcagttt ggaccggaaa tagttcttga agccttcaac aaacactctc atgagaacat
     3421 caacaaggaa gaagagtgct atagccaggg aaacaccttc aagaatctct ctaatgcctc
     3481 ttttcttctc actgatggcc agatccacaa tcaccactat gatgtctaca atgataagca
     3541 caactccaaa tactctgaat ccaaatgaca tcacaaacgg ggaaattttt ctcctaatgg
     3601 tattggaaca gctgtcttgt tcattaccct catcacgacc atcatcaatc tttatcgtgg
     3661 gctcatctgc tgttggagag ctcgaagttt tagtgagctc tgacccctgt tcatacctca
     3721 cagtcgtctc catggtggcg ctagcataac ttcgtataaa gtatcctata cgaagttatt
     3781 tgccttaacc cagaaattat cactgttatt ctttagaatg gtgcaaagaa taacttcgta
     3841 taatgtatgc tatacgaagt tatgaattcg atatcaagct tatcgataat caacctctgg
     3901 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     3961 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     4021 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     4081 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     4141 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     4201 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     4261 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     4321 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     4381 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     4441 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga
     4501 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     4561 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     4621 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     4681 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     4741 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     4801 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     4861 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     4921 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     4981 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     5041 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     5101 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     5161 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     5221 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5281 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     5341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     5401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     5461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     5581 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     5641 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     5701 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     5761 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5821 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5881 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     5941 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     6001 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     6061 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     6121 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     6181 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     6241 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     6301 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     6361 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     6421 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     6481 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     6541 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     6601 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     6661 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     6721 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     6781 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     6841 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     6901 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     6961 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     7021 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     7081 cagaccccgt agaaaagatc aaaggatctt c
//
