LOCUS       sequence-353674-        6898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Double floxed genetically encoded voltage indicator (GEVI) JEDI-2P 
            in AAV production vector under the control of the mammalian promoter
            (EF1a).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6898)
  AUTHORS   Liu Z, Lu X, Villette V, Gou Y, Colbert KL, Lai S, Guan S, Land MA, 
            Lee J, Assefa T, Zollinger DR, Korympidou MM, Vlasits AL, Pang MM, 
            Su S, Cai C, Froudarakis E, Zhou N, Patel SS, Smith CL, Ayon A, 
            Bizouard P, Bradley J, Franke K, Clandinin TR, Giovannucci A, Tolias
            AS, Reimer J, Dieudonne S, St-Pierre F
  TITLE     Sustained deep-tissue voltage recording using a fast indicator 
            evolved for two-photon microscopy.
  JOURNAL   Cell. 2022 Aug 16. pii: S0092-8674(22)00916-3. doi: 
            10.1016/j.cell.2022.07.013.
  PUBMED    35985322
REFERENCE   2  (bases 1 to 6898)
  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. 2022 
            Aug 16. pii: S0092-8674(22)00916-3. doi: 
            10.1016/j.cell.2022.07.013."
COMMENT     Sequence Label: pAAV-EF1a-DIO-JEDI-2P-WPRE
FEATURES             Location/Qualifiers
     source          1..6898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      326..335
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    346..379
                     /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    430..463
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    488..1076
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(541..561)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(959..970)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1108..1584
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(1307..1326)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   1624..1764
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1624..1753
                     /label=AAV2 ITR
     rep_origin      1839..2294
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1926..1945)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2136..2157
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2311..2330)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2430..2452
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2490..2508)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2576..2680
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2681..2749,2750..3541)
                     /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(2899..2918)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3712..4300
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4201..4220
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4362..4491
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        4592..5770
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4823..5761
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5718..5738
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(5801..5834)
                     /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(5885..5918)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 actctcatga gaacatcaac aaggaagaag agtgctatag ccagggaaac accttcaaga
       61 atctctctaa tgcctctttt cttctcactg atggccagat ccacaatcac cactatgatg
      121 tctacaatga taagcacaac tccaaatact ctgaatccaa atgacatcac aaacggggaa
      181 atttttctcc taatggtatt ggaacagctg tcttgttcat taccctcatc acgaccatca
      241 tcaatcttta tcgtgggctc atctgctgtt ggagagctcg aagttttagt gagctctgac
      301 ccctgttcat acctcacagt cgtctccatg gtggcggtag ctagcataac ttcgtataaa
      361 gtatcctata cgaagttatt tgccttaacc cagaaattat cactgttatt ctttagaatg
      421 gtgcaaagaa taacttcgta taatgtatgc tatacgaagt tatgaattcg atatcaagct
      481 tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta
      541 tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc
      601 ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga
      661 ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac
      721 ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc
      781 cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc
      841 tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg
      901 gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc
      961 ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc
     1021 gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg
     1081 ataccgagcg ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct
     1141 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt
     1201 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta
     1261 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca
     1321 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg
     1381 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc
     1441 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac
     1501 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga
     1561 accactgctc ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc
     1621 cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg
     1681 aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg
     1741 agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt
     1801 gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt
     1861 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc
     1921 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     1981 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2041 caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2101 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2161 aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt
     2221 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     2281 aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     2341 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2401 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     2461 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     2521 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     2581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     2641 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     2701 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2761 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2821 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2881 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2941 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3001 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3061 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     3121 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     3181 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     3241 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     3301 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     3361 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     3421 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     3481 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     3541 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     3601 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     3661 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     3721 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     3781 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     3841 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     3901 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     3961 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4021 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4081 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     4141 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     4201 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     4261 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     4321 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
     4381 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
     4441 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
     4501 cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg
     4561 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
     4621 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
     4681 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
     4741 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
     4801 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
     4861 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
     4921 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
     4981 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
     5041 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
     5101 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
     5161 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
     5221 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
     5281 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
     5341 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
     5401 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     5461 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     5521 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     5581 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     5641 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     5701 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     5761 ggtgtcgtga ggtaccggat cctctagagt cgactccgga ataacttcgt ataggatact
     5821 ttatacgaag ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct
     5881 cttaataact tcgtatagca tacattatac gaagttatgg cgcgccctat tacttgtaca
     5941 tcattaggtt accacttcaa gttgtttctt ctgtgaagcc aggcgaaata ttcttattaa
     6001 gataacaatt ttcagaactc gaagaagagt aaccatgtgc ggcatctgat ctgtatcata
     6061 ttccagttta tgccccagaa tgttgccatc ttctttaaaa tcggtgcctt tcagttcaat
     6121 gcggttcacc agggtatcgc cttcaaattt caccaccgcg cgggttttat atttgccatc
     6181 atctttaaag ctaatggtgc gttcctgcac atagccttcc ggcatcgcgc ttttaaaaaa
     6241 atcatggcgt ttcatatgat ccggatagcg gctaaagcac tgcacgccat aggttaaggt
     6301 ggtcaccagg gtcggccacg gcaccggtag tttgccggtg gtgcaaataa atttcagggt
     6361 cagtttgcca atggtcgcat cgccttcgcc ttcgccgcgc acgctaaatt tatggccgtt
     6421 cacatcgcca tccagttcca ccagaatcgg caccacgccg gtaaacagtt cttcgccctg
     6481 gctcgcgctg ccgccggtgc cgccatacag ttcatccatg ccgtgtgtaa tgcccgctgc
     6541 ggtcacaaat tccagcagca ccatgtgatc gcgtttttcg ttcggatctt tgctcagcac
     6601 ggtctggtgg ctcagataat ggttatccgg cagcagcacc gggccatcgc caatcggggt
     6661 gttctgctga taatgatccg ccagctgcac gctgccatct tccacgttat ggcgcacggt
     6721 aaagttcgct ttaatgccgt ttttctgttt atccgcggta atatacacgt tctcgctgtt
     6781 aaaggcagca aggtcagaga aggagtaggt catattaatt agcagagtgc ccactactat
     6841 gactgcatcc aaagtaacca gtttggaccg gaaatagttc ttgaagcctt caacaaac
//
