LOCUS       sequence-265918-        6877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses ASAP3(ASAP-family genetically encoded voltage indicator) 
            in cre recombinase expressing mouse/cell, gene delivered by 
            pAAV-EF1a vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6877)
  AUTHORS   Villette V, Chavarha M, Dimov IK, Bradley J, Pradhan L, Mathieu B, 
            Evans SW, Chamberland S, Shi D, Yang R, Kim BB, Ayon A, Jalil A, 
            St-Pierre F, Schnitzer MJ, Bi G, Toth K, Ding J, Dieudonne S, Lin MZ
  TITLE     Ultrafast Two-Photon Imaging of a High-Gain Voltage Indicator in 
            Awake Behaving Mice.
  JOURNAL   Cell. 2019 Dec 12;179(7):1590-1608.e23. doi: 
            10.1016/j.cell.2019.11.004.
  PUBMED    31835034
REFERENCE   2  (bases 1 to 6877)
  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: 
            "10.1016/j.cell.2019.11"; journalName: "Cell"; date: "2019-12-12- 
            12"; volume: "179"; issue: "7"; pages: "1590-1608.e23"
COMMENT     Sequence Label: pAAV-EF1a-DIO-ASAP3-WPRE
FEATURES             Location/Qualifiers
     source          1..6877
                     /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        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1440..1473)
                     /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(1524..1557)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2846..2855
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2862..2895
                     /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    2946..2979
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3004..3592
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3057..3077)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3475..3486)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3624..4100
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3823..3842)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4140..4280
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4140..4269
                     /label=AAV2 ITR
     rep_origin      4355..4810
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4442..4461)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4652..4673
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4827..4846)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4946..4968
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5006..5024)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5092..5196
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5197..5265,5266..6057)
                     /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(5415..5434)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6228..6816
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6717..6736
                     /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 gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa
     1441 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
     1501 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
     1561 gcgcctcatt aggttaccac ttcaagttgt ttcttctgtg aagccagctg aaatattctt
     1621 attaagataa caattctcag aactcgaaga agagtaacca tctgcggcat ctgatctgtg
     1681 ttatattcca gtttatgccc cagaatgttg ccatcttctt taaaatcggt gcctttcagt
     1741 tcaatgcggt tcaccagggt atcgccttca aatttcacca ccgcgcgggt tttatatttg
     1801 ccatcatctt taaagctaat ggtgcgttcc tgcacatagc cttccggcat cgcgctttta
     1861 aaaaaatcat ggcgtttcat atgatccgga tagcggctaa agcactgcac gccataggtt
     1921 aaggtggtca ccagggtcgg ccacggcacc ggtagtttgc cggtggtgca aataaatttc
     1981 agggtcagtt tgccaatggt cgcatcgcct tcgccttcgc cgcgcacgct aaatttatgg
     2041 ccgttcacat cgccatccag ttccaccaga atcggcacca cgccggtaaa cagttcttcg
     2101 ccctggctcg cgctgccgcc ggtgccgcca tacagttcat ccatgccgtg tgtaatgccc
     2161 gctgcggtca caaattccag cagcaccatg tgatcgcgtt tttcgttcgg atctttgctc
     2221 agcacggtct gggtgctcag ataatggtta tccggcagca gcaccgggcc atcgccaatc
     2281 ggggtgttct gctgataatg atccgccagc tgcacgctgc catcttccac gttatggcgc
     2341 acggtaaagt tcgctttaat gccgtttttc tgtttatccg cggtaatata cacgttatca
     2401 ccgcgaaatg taccgtcaga gaaggagtag gtcatattaa ttagcagagt gcccactact
     2461 atgactgcat ccaaagtatt cagtttggac cggaaatagt tcttgaagcc ttcaacaaac
     2521 actctcatga gaacatcaac aaggaagaag agtgctatag ccagggaaac accttcaaga
     2581 atctctctaa tgcctctttt cttctcactg atggccagat ccacaatcac cactatgatg
     2641 tctacaatga taagcacaac tccaaatact ctgaatccaa atgacatcac aaacggggaa
     2701 atttttctcc taatggtatt ggaacagctg tcttgttcat taccctcatc acgaccatca
     2761 tcaatcttta tcgtgggctc atctgctgtt ggagagctcg aagttttagt gagctctgac
     2821 ccctgttcat acctcacagt cgtctccatg gtggcgctag cataacttcg tataaagtat
     2881 cctatacgaa gttatttgcc ttaacccaga aattatcact gttattcttt agaatggtgc
     2941 aaagaataac ttcgtataat gtatgctata cgaagttatg aattcgatat caagcttatc
     3001 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3061 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3121 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3181 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3241 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3301 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3361 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     3421 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3481 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3541 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     3601 cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     3661 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     3721 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     3781 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     3841 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     3901 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     3961 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     4021 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     4081 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     4141 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4201 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4261 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4321 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4381 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     4441 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4501 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4561 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4621 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4681 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4741 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4801 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4861 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4921 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4981 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5041 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5101 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5161 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5221 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5281 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5341 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5401 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5461 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5521 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5581 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5641 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5701 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5761 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5821 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     5881 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     5941 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6001 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6061 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6121 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6181 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6241 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6301 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6361 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6421 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6481 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6541 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6601 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6661 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6721 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6781 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6841 cggttcctgg ccttttgctg gccttttgct cacatgt
//
