LOCUS       sequence-414781-        6430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Inducing apoptotic ablation of neurons through a process reliant on 
            Flp recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6430)
  AUTHORS   Goto T, Hagihara M, Irie S, Abe T, Kiyonari H, Miyamichi K
  TITLE     Dietary availability acutely influences puberty onset via a 
            hypothalamic neural circuit.
  JOURNAL   Neuron. 2025 Feb 17:S0896-6273(25)00041-8. doi: 
            10.1016/j.neuron.2025.01.015.
  PUBMED    39999843
REFERENCE   2  (bases 1 to 6430)
  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: "Neuron. 
            2025 Feb 17:S0896-6273(25)00041-8. doi: 
            10.1016/j.neuron.2025.01.015."
COMMENT     Sequence Label: pAAV hSyn-fDIO-taCasp3-TEVp
FEATURES             Location/Qualifiers
     source          1..6430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        121..568
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    606..639
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     CDS             complement(1437..1490)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(1806..1826)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     regulatory      2345..2354
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2361..2394)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    2503..3091
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2556..2576)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2974..2985)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3123..3599
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3322..3341)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3639..3779
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3639..3768
                     /label=AAV2 ITR
     rep_origin      3854..4309
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3941..3960)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4151..4172
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4326..4345)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4445..4467
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4505..4523)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4591..4695
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4696..4764,4765..5556)
                     /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(4914..4933)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5727..6315
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6216..6235
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6377..6430,1..76)
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
       61 atcactaggg gttcctgcgg ccgcacgcgt gtgtctagac tgcagagggc cctgcgtatg
      121 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      181 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      241 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      301 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      361 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      421 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      481 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      541 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac cggatcctct agagtcgact
      601 ccggagaagt tcctattctc tagaaagtat aggaacttcg cagaatggta gctggattgt
      661 agctgctatt agcaatatga aacctcttag aagttcctat tcttcaaaag gtataggaac
      721 ttcggcgcgc ctcaaagctg agtggcttcc ttaacgggct ggaagggttc ctcgggcttg
      781 accatgaaca ccttatgtcc gccccaaaga acgctgtcgg cgttcagcct ccagccgctc
      841 acccactgct gcgcctcttg attggtcaac agctccatga agtttttcgg cacgctcgtg
      901 aagtagttgt tggtattggt aaaattgctg gcgctgtgga tgcccacgat gaagccgtcc
      961 ctagtgctca ccaatgggct gccgcattgg ccatccttcg tctgaatcca gtgcttccag
     1021 aagatgccgt cgctgctggg gaaggtgcag ctggtatcag agaccatgct gctcatggac
     1081 ttggtctgga agttggtggt caccaggcag attcgctcct ccctttgtgg ctccctgaac
     1141 ttcagtttct ggggaaaggg ggggaagtcc ttgggcatcc tgatgataat catgtccctg
     1201 ccgtcgatca ggtgctgttg cagggtggtt gtgttcttca ctttgaacac gccgtgcagg
     1261 ctttgcacca gcagtgtgcc gttattcctc ctgaacagat gcttgttggt gatgatgaag
     1321 gggccgaagc cgatgccgta cagactagtg gtgtgaccgt cgctctcgtt ggtcaggtga
     1381 cagatggtgc tggagatcgg gttgtagtcc ctcgggccct tgaaaaggct ctcgccaggg
     1441 ccgggattct cctccacgtc accgcatgtt agaagacttc ctctgccctc tccgcttccg
     1501 gagtgataaa aatagagttc ttttgtgagc atggaaacaa tacatggaat ctgtttcttt
     1561 gcatgaaaag tagcgtcaaa ggaaaaggac tcaaattctg ttgccacctt tcggttaacc
     1621 cgggtaagaa tgtgcataaa ttcaagcttg tcggcatact gtttcagcat ggcacaaagc
     1681 gactggatga accaggagcc atcctttgaa tttcgccaag aataataacc aggtgctgtg
     1741 gagtatgcat acaagaagtc ggcctccact ggtattttat gacacgccat gtcatcatca
     1801 acaccactct ggaagtagag attttctgtc tcaatgccac agtccagttc tgtaccacgg
     1861 caggcctgaa taatgaaaag tttgggtttt ccagttagac ttctacaacg atcccctctg
     1921 aaaaagtttg ttattttttt caggtcaaca ggtccatttg ttccaaaaat tattccttct
     1981 tcaccatggc tcagaagcac acaaacaaaa ctgctccttt tgctgtgatc ttctttagaa
     2041 acatcacgca tcaattccac aatttcttca cgtgtaagat catttttatt cctgacttca
     2101 tatttcaagt ttctgaatgt ttccctgagg tttgctgcat cgacatctgt accagaccga
     2161 gatgtcattc cagtgctttt atgaaaattc ttattattaa ttattataca taaacccatc
     2221 tcaggataat ccattttata actgttgtcc agggatattc cagagtccat tgattcgctt
     2281 ccatgtatga tctttggttc caaattttta atggattttg aatccactga gttttcagtg
     2341 ttctccatgg tggcgctagc gaagttccta tactttctag agaataggaa cttcttgcct
     2401 taacccagaa attatcactg ttattcttta gaatggtgca aagagaagtt cctatacctt
     2461 ttgaagaata ggaacttcga attcgatatc aagcttatcg ataatcaacc tctggattac
     2521 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     2581 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     2641 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     2701 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     2761 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     2821 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2881 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg
     2941 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     3001 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     3061 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct
     3121 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc
     3181 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt
     3241 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga
     3301 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt
     3361 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt
     3421 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac
     3481 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac
     3541 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc
     3601 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt
     3661 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     3721 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     3781 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     3841 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3901 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     4201 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     4261 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     4321 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     4381 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     4441 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     4501 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4561 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     4621 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     4681 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     4741 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     4801 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     4861 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     4921 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4981 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5041 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5101 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5161 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5221 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5281 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5341 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5401 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5461 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5521 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     5581 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5641 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5701 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5761 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5821 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5881 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5941 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6001 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6061 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6121 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6181 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6241 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6301 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6361 ccttttgctc acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg
     6421 gcgtcgggcg
//
