LOCUS       sequence-491262-        6904 bp    DNA     circular SYN 31-OCT-2025
DEFINITION  Double floxed genetically encoded voltage indicator (GEVI) 
            JEDI-2Psub 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 6904)
  TITLE     All-optical voltage interrogation for probing synaptic plasticity in
            vivo
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6904)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Oct 31, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-EF1a-DIO-JEDI-2Psub-WPRE
FEATURES             Location/Qualifiers
     source          1..6904
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        232..1410
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          463..1401
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1358..1378
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1441..1474)
                     /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(1525..1558)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2870..2879
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2890..2923
                     /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    2974..3007
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3032..3620
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3085..3105)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3503..3514)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3652..4128
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3851..3870)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4168..4308
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4168..4297
                     /label=AAV2 ITR
     rep_origin      4383..4838
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4470..4489)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4680..4701
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4855..4874)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4974..4996
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5034..5052)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5120..5224
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5225..5293,5294..6085)
                     /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(5443..5462)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6256..6844
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6745..6764
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag
      181 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
      241 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
      301 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
      361 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
      421 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
      481 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
      541 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      601 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
      661 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
      721 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
      781 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
      841 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
      901 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
      961 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     1021 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     1081 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1141 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1201 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1261 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1321 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1381 agtttttttc ttccatttca ggtgtcgtga ggtaccggat cctctagagt cgactccgga
     1441 ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg attgtagctg
     1501 ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac gaagttatgg
     1561 cgcgccctat tacttgtaca ggtaccttag gttaccactt caagttgttt cttctgtgaa
     1621 gccaggcgaa atattcttat taagataaca attttcagaa ctcgaagaag agtaaccatg
     1681 tgcggcatct gatctgtcca atcatattcc agtttatgcc ccagaatgtt gccatcttct
     1741 ttaaaatcgg tgcctttcag ttcaatgcgg ttcaccaggg tatcgccttc aaatttcacc
     1801 accgcgcggg ttttatattt gccatcatct ttaaagctaa tggtgcgttc ctgcacatag
     1861 ccttccggca tcgcgctttt aaaaaaatca tggcgtttca tatgatccgg atagcggcta
     1921 aagcactgca cgccataggt taaggtggtc accagggtcg gccacggcac cggtagtttg
     1981 ccggtggtgc aaataaattt cagggtcagt ttgccaatgg tcgcatcgcc ttcgccttcg
     2041 ccgcgcacgc taaatttatg gccgttcaca tcgccatcca gttccaccag aatcggcacc
     2101 acgccggtaa acagttcttc gccctggctc gcgctgccgc cggtgccgcc atacagttca
     2161 tccatgccgt gtgtaatgcc cgctgcggtc acaaattcca gcagcaccat gtgatcgcgt
     2221 ttttcgttcg gatctttgct cagcacggtc tggtggctca gataatggtt atccggcagc
     2281 agcaccgggc catcgccaat cggggtgttc tgctgataat gatccgccag ctgcacgctg
     2341 ccatcttcca cgttatggcg cacggtaaag ttcgctttaa tgccgttttt ctgtttatcc
     2401 gcggtaatat acacgttctc gctgttaaag gcagcaaggt cagagaagga gtaggtcata
     2461 ttaattagca gagtgcccac tactatgact gcatccaaag taaccagttt ggaccggaaa
     2521 tagttcttga agccttcaac aaacactctc atgagaacat caacaaggaa gaagagtgct
     2581 atagccaggg aaacaccttc aagaatctct ctaatgcctc ttttcttctc actgatggcc
     2641 agatccacaa tcaccactat gatgtctaca atgataagca caactccaaa tactctgaat
     2701 ccaaatgaca tcacaaacgg ggaaattttt ctcctaatgg tattggaaca gctgtcttgt
     2761 tcattaccct catcacgacc atcatcaatc tttatcgtgg gctcatctgc tgttggagag
     2821 ctcgaagttt tagtgagctc tgacccctgt tcatacctca cagtcgtctc catggtggcg
     2881 gtagctagca taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa
     2941 ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg
     3001 aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga
     3061 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     3121 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     3181 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     3241 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     3301 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     3361 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     3421 ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg
     3481 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     3541 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     3601 cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat
     3661 ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc
     3721 agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat
     3781 attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg
     3841 gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca
     3901 atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc
     3961 caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc
     4021 atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc
     4081 aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag
     4141 gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct
     4201 ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct
     4261 ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc
     4321 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata
     4381 gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     4441 cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     4501 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     4561 tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg
     4621 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     4681 tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt
     4741 ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt
     4801 taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat
     4861 ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc
     4921 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     4981 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt
     5041 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     5101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     5161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     5221 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     5281 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     5341 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     5401 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     5461 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     5521 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     5581 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     5641 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     5701 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     5761 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     5821 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     5881 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     5941 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     6001 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     6061 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     6121 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     6181 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     6241 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     6301 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     6361 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     6421 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6481 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6541 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6601 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6661 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6721 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6781 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6841 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6901 catg
//
