LOCUS       sequence-491263-        7135 bp    DNA     circular SYN 31-OCT-2025
DEFINITION  Double floxed soma and AIS-localized genetically encoded voltage 
            indicator (GEVI) JEDI-2Psub in AAV production vector expressed under
            the mammalian promoter (EF1a).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7135)
  TITLE     All-optical voltage interrogation for probing synaptic plasticity in
            vivo
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7135)
  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            "Unpublished"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-Kv-WPRE
FEATURES             Location/Qualifiers
     source          1..7135
                     /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      3101..3110
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3121..3154
                     /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    3205..3238
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3263..3851
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3316..3336)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3734..3745)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3883..4359
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4082..4101)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4399..4539
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4399..4528
                     /label=AAV2 ITR
     rep_origin      4614..5069
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4701..4720)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4911..4932
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5086..5105)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5205..5227
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5265..5283)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5351..5455
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5456..5524,5525..6316)
                     /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(5674..5693)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6487..7075
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6976..6995
                     /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 ttatcaaaag cgggtcgctt ccggaaaatc ggtcgcgcag
     1621 ctaataaagc tatcaatgct gctcatgctg cgcatatcaa tcacgccttc ggtgcgcgcc
     1681 ggcagcggcg ccaccgggct cggcatgctg ctcatttcca gttcttccgg cggtttgctc
     1741 tgcggcgcca tttctttggt gttcagaatc ggctggctct gacgcgtgga agatccagat
     1801 gaacctgaag aacccttaag ggttaccact tcaagttgtt tcttctgtga agccaggcga
     1861 aatattctta ttaagataac aattttcaga actcgaagaa gagtaaccat gtgcggcatc
     1921 tgatctgtcc aatcatattc cagtttatgc cccagaatgt tgccatcttc tttaaaatcg
     1981 gtgcctttca gttcaatgcg gttcaccagg gtatcgcctt caaatttcac caccgcgcgg
     2041 gttttatatt tgccatcatc tttaaagcta atggtgcgtt cctgcacata gccttccggc
     2101 atcgcgcttt taaaaaaatc atggcgtttc atatgatccg gatagcggct aaagcactgc
     2161 acgccatagg ttaaggtggt caccagggtc ggccacggca ccggtagttt gccggtggtg
     2221 caaataaatt tcagggtcag tttgccaatg gtcgcatcgc cttcgccttc gccgcgcacg
     2281 ctaaatttat ggccgttcac atcgccatcc agttccacca gaatcggcac cacgccggta
     2341 aacagttctt cgccctggct cgcgctgccg ccggtgccgc catacagttc atccatgccg
     2401 tgtgtaatgc ccgctgcggt cacaaattcc agcagcacca tgtgatcgcg tttttcgttc
     2461 ggatctttgc tcagcacggt ctggtggctc agataatggt tatccggcag cagcaccggg
     2521 ccatcgccaa tcggggtgtt ctgctgataa tgatccgcca gctgcacgct gccatcttcc
     2581 acgttatggc gcacggtaaa gttcgcttta atgccgtttt tctgtttatc cgcggtaata
     2641 tacacgttct cgctgttaaa ggcagcaagg tcagagaagg agtaggtcat attaattagc
     2701 agagtgccca ctactatgac tgcatccaaa gtaaccagtt tggaccggaa atagttcttg
     2761 aagccttcaa caaacactct catgagaaca tcaacaagga agaagagtgc tatagccagg
     2821 gaaacacctt caagaatctc tctaatgcct cttttcttct cactgatggc cagatccaca
     2881 atcaccacta tgatgtctac aatgataagc acaactccaa atactctgaa tccaaatgac
     2941 atcacaaacg gggaaatttt tctcctaatg gtattggaac agctgtcttg ttcattaccc
     3001 tcatcacgac catcatcaat ctttatcgtg ggctcatctg ctgttggaga gctcgaagtt
     3061 ttagtgagct ctgacccctg ttcatacctc acagtcgtct ccatggtggc ggtagctagc
     3121 ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa attatcactg
     3181 ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac gaagttatga
     3241 attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac
     3301 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3361 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3421 gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt
     3481 gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg
     3541 gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg
     3601 ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc
     3661 atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt
     3721 ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc
     3781 tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc
     3841 cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac
     3901 ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc
     3961 ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg
     4021 tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg
     4081 gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc
     4141 tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc
     4201 atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc
     4261 aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg
     4321 ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg
     4381 tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4441 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4501 cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc
     4561 tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc
     4621 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4681 tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4741 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4801 acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc
     4861 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4921 gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat
     4981 tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     5041 ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga
     5101 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     5161 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     5221 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     5281 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5341 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5401 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5461 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5521 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5581 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5641 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5701 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5761 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5821 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5881 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     5941 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6001 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6061 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6121 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6181 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6241 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6301 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6361 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6421 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6481 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6541 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6601 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     6661 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6721 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6781 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6841 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     6901 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     6961 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7021 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7081 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatg
//
