LOCUS       sequence-477099-        6710 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Constitutive expression of genetically encoded voltage indicator 
            ASAP3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6710)
  AUTHORS   Haziza S, Chrapkiewicz R, Zhang Y, Kruzhilin V, Li J, Li J, Delamare
            G, Swanson R, Buzsaki G, Kannan M, Vasan G, Lin MZ, Zeng H, Daigle 
            TL, Schnitzer MJ
  TITLE     Imaging high-frequency voltage dynamics in multiple neuron classes 
            of behaving mammals.
  JOURNAL   Cell, 188, 1-23, August 7, 2025. doi: 10.1016/j.cell.2025.06.028
  PUBMED    39185175
REFERENCE   2  (bases 1 to 6710)
  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: "Cell, 188, 
            1-23, August 7, 2025. doi: 10.1016/j.cell.2025.06.028"
COMMENT     Sequence Label: pAAV-CaMKII-ASAP3-WPRE
FEATURES             Location/Qualifiers
     source          1..6710
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        10..1298
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1323..1332
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1338..1355
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      1368..1377
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2698..3286
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2751..2771)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3169..3180)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3318..3794
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3517..3536)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3834..3974
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3834..3963
                     /label=AAV2 ITR
     rep_origin      4049..4504
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4136..4155)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4346..4367
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4521..4540)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4640..4662
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4700..4718)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4786..4890
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4891..4959,4960..5751)
                     /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(5109..5128)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5922..6510
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6411..6430
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6572..6701
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 cgcgtttaac attatggcct taggtcactt catctccatg gggttcttct tctgattttc
       61 tagaaaatga gatgggggtg cagagagctt cctcagtgac ctgcccaggg tcacatcaga
      121 aatgtcagag ctagaacttg aactcagatt actaatctta aattccatgc cttgggggca
      181 tgcaagtacg atatacagaa ggagtgaact cattagggca gatgaccaat gagtttagga
      241 aagaagagtc cagggcaggg tacatctaca ccacccgccc agccctgggt gagtccagcc
      301 acgttcacct cattatagtt gcctctctcc agtcctacct tgacgggaag cacaagcaga
      361 aactgggaca ggagccccag gagaccaaat cttcatggtc cctctgggag gatgggtggg
      421 gagagctgtg gcagaggcct caggaggggc cctgctgctc agtggtgaca gataggggtg
      481 agaaagcaga cagagtcatt ccgtcagcat tctgggtctg tttggtactt cttctcacgc
      541 taaggtggcg gtgtgatatg cacaatggct aaaaagcagg gagagctgga aagaaacaag
      601 gacagagaca gaggccaagt caaccagacc aattcccaga ggaagcaaag aaaccattac
      661 agagactaca agggggaagg gaaggagaga tgaattagct tcccctgtaa accttagaac
      721 ccagctgttg ccagggcaac ggggcaatac ctgtctcttc agaggagatg aagttgccag
      781 ggtaactaca tcctgtcttt ctcaaggacc atcccagaat gtggcaccca ctagccgtta
      841 ccatagcaac tgcctctttg ccccacttaa tcccatcccg tctgttaaaa gggccctata
      901 gttggaggtg ggggaggtag gaagagcgat gatcacttgt ggactaagtt tgttcgcatc
      961 cccttctcca accccctcag tacatcaccc tgggggaaca gggtccactt gctcctgggc
     1021 ccacacagtc ctgcagtatt gtgtatataa ggccagggca aagaggagca ggttttaaag
     1081 tgaaaggcag gcaggtgttg gggaggcagt taccggggca acgggaacag ggcgtttcgg
     1141 aggtggttgc catggggacc tggatgctga cgaaggctcg cgaggctgtg agcagccaca
     1201 gtgccctgct cagaagcccc aagctcgtca gtcaagccgg ttctccgttt gcactcagga
     1261 gcacgggcag gcgagtggcc cctagttctg ggggcagctc tagagcggta ccggatccac
     1321 ccgccaccat gggttctcat catcatcatc atcatggtat ggctagcgcc accatggaga
     1381 cgactgtgag gtatgaacag gggtcagagc tcactaaaac ttcgagctct ccaacagcag
     1441 atgagcccac gataaagatt gatgatggtc gtgatgaggg taatgaacaa gacagctgtt
     1501 ccaataccat taggagaaaa atttccccgt ttgtgatgtc atttggattc agagtatttg
     1561 gagttgtgct tatcattgta gacatcatag tggtgattgt ggatctggcc atcagtgaga
     1621 agaaaagagg cattagagag attcttgaag gtgtttccct ggctatagca ctcttcttcc
     1681 ttgttgatgt tctcatgaga gtgtttgttg aaggcttcaa gaactatttc cggtccaaac
     1741 tgaatacttt ggatgcagtc atagtagtgg gcactctgct aattaatatg acctactcct
     1801 tctctgacgg tacatttcgc ggtgataacg tgtatattac cgcggataaa cagaaaaacg
     1861 gcattaaagc gaactttacc gtgcgccata acgtggaaga tggcagcgtg cagctggcgg
     1921 atcattatca gcagaacacc ccgattggcg atggcccggt gctgctgccg gataaccatt
     1981 atctgagcac ccagaccgtg ctgagcaaag atccgaacga aaaacgcgat cacatggtgc
     2041 tgctggaatt tgtgaccgca gcgggcatta cacacggcat ggatgaactg tatggcggca
     2101 ccggcggcag cgcgagccag ggcgaagaac tgtttaccgg cgtggtgccg attctggtgg
     2161 aactggatgg cgatgtgaac ggccataaat ttagcgtgcg cggcgaaggc gaaggcgatg
     2221 cgaccattgg caaactgacc ctgaaattta tttgcaccac cggcaaacta ccggtgccgt
     2281 ggccgaccct ggtgaccacc ttaacctatg gcgtgcagtg ctttagccgc tatccggatc
     2341 atatgaaacg ccatgatttt tttaaaagcg cgatgccgga aggctatgtg caggaacgca
     2401 ccattagctt taaagatgat ggcaaatata aaacccgcgc ggtggtgaaa tttgaaggcg
     2461 ataccctggt gaaccgcatt gaactgaaag gcaccgattt taaagaagat ggcaacattc
     2521 tggggcataa actggaatat aacacagatc agatgccgca gatggttact cttcttcgag
     2581 ttctgagaat tgttatctta ataagaatat ttcagctggc ttcacagaag aaacaacttg
     2641 aagtggtaac ctaatgaggc gcgcccctgc agggaattcg atatcaagct tatcgataat
     2701 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     2761 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     2821 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     2881 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     2941 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3001 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3061 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     3121 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3181 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3241 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg
     3301 ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     3361 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     3421 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     3481 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     3541 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     3601 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     3661 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     3721 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     3781 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc
     3841 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     3901 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     3961 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     4021 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     4081 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     4141 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4201 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4261 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4321 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4381 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     4441 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     4501 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     4561 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4621 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4681 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     4741 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4801 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4861 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4921 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4981 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     5041 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     5101 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     5161 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5221 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5281 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5341 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5401 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5461 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5521 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5581 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5641 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5701 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5761 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5821 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5881 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5941 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     6001 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     6061 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     6121 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6181 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6241 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6301 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6361 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6421 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6481 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6541 ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc
     6601 actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc
     6661 gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
//
