LOCUS       sequence-444378-        7534 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses vTRAP and hM3D(Gq) in a FlpON-CreOFF dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7534)
  AUTHORS   Selten M, Bernard C, Mukherjee D, Hamid F, Hanusz-Godoy A, Oozeer F,
            Zimmer C, Marin O
  TITLE     Regulation of PV interneuron plasticity by neuropeptide-encoding 
            genes.
  JOURNAL   Nature. 2025 Apr 30. doi: 10.1038/s41586-025-08933-z.
  PUBMED    40307547
REFERENCE   2  (bases 1 to 7534)
  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: "Nature. 
            2025 Apr 30. doi: 10.1038/s41586-025-08933-z."
COMMENT     Sequence Label: pAAV-hSyn-flx-fDIO-HA-Rpl10a-T2A-myc-hM3D(Gq)
FEATURES             Location/Qualifiers
     source          1..7534
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(21..32)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    170..646
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(369..388)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   686..826
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   686..815
                     /label=AAV2 ITR
     rep_origin      901..1356
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(988..1007)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1198..1219
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1373..1392)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1492..1514
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        1638..1742
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1743..1811,1812..2603)
                     /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(1961..1980)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2774..3362
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3263..3282
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3424..3553
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        3598..4045
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(4065..4098)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    4115..4162
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(4289..4318)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(6089..6142)
                     /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(6791..6817)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    complement(6840..6887)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    complement(7026..7059)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    join(7084..7534,1..138)
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(7137..7157)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
       61 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
      121 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc
      181 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag
      241 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat
      301 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc
      361 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat
      421 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca
      481 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat
      541 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa
      601 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt
      661 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct
      721 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt
      781 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg
      841 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt
      901 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      961 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1021 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1081 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc
     1141 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1201 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat
     1261 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta
     1321 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct
     1381 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct
     1441 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     1501 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga
     1561 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     1621 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     1681 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     1741 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     1801 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     1861 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     1921 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     1981 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     2041 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     2101 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     2161 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     2221 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     2281 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     2341 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     2401 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     2461 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     2521 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     2581 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     2641 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     2701 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     2761 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     2821 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     2881 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     2941 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     3001 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     3061 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     3121 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     3181 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     3241 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     3301 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     3361 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     3421 tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc
     3481 tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc caactccatc
     3541 actaggggtt cctgcggccg cacgcgtgtg tctagactgc agagggccct gcgtatgagt
     3601 gcaagtgggt tttaggacca ggatgaggcg gggtgggggt gcctacctga cgaccgaccc
     3661 cgacccactg gacaagcacc caacccccat tccccaaatt gcgcatcccc tatcagagag
     3721 ggggagggga aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac
     3781 agtgccttcg cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct
     3841 gacgtcactc gccggtcccc cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc
     3901 cgccgccggc ccagccggac cgcaccacgc gaggcgcgag ataggggggc acgggcgcga
     3961 ccatctgcgc tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg gcagcggagg
     4021 agtcgtgtcg tgcctgagag cgcagtcgag aaggtaccgg atccataact tcgtatagca
     4081 tacattatac gaagttatat accgcgggcc cggggaagtt cctattccga agttcctatt
     4141 ctctagaaag tataggaact tcatcaaaat aggaagacca atgcttcacc atcgacccga
     4201 attgccaagc atcaccatcg acagaagttc ctattccgaa gttcctattc ttcaaaaggt
     4261 ataggaactt cgtcgacggc gcgccttaca gatcttcttc agagatgagt ttctgctcct
     4321 tcaaggcctg ctcgggtgcg cgcttgtgaa aaatgaccga ctgtctctgc tggtactgct
     4381 gcttgcgcct cttttttttg tcacactggc acagcagcag catcttgaaa gtggttctga
     4441 atgttttgtt gcacagagca tagcacacgg ggttcacggt gctgttgatg tagcacagcc
     4501 agtagcccag attccaaaag gttttgggta tgcagctgtc acaaaaggtg ttcaccagaa
     4561 ccatgatgtt gtatggggtc caagtgatga tgaaggcaag caagatcgca ctgagggtct
     4621 gggccgcttt cttctccttg accagggaca tccttttccg cttagtgatc tgacttctgg
     4681 tcttcagagc aaacctcttg gccagagtgg cttccttgaa ggacagaggt agagtggccg
     4741 tgctcttacc cactgaggag ttgacgtcag aagtcttagc tgtgtccacg gctgactcta
     4801 gctggatggg aagcttggag aagctttttg gaaaactgcc tccatcgtcc acgctcttct
     4861 gggcctgcag cttgtcggct ttcctctcca agtccaccat ccccagctcc tcctcaggca
     4921 cctgcaggtt gtccgatgag ggtaacttgg tggagttgag gatggtgctg tgacccggaa
     4981 gcttgagcac gatggagtag atggctctcg tctcggagcc aatgtcctcc tcgtcggagg
     5041 aggcggagtt ctccagggag gcagcagcat cattgttgtt ccaactgtca ctgctgctgt
     5101 ggtcttggtc catctgctcg gagctgggtt tccagctctt ggttgtgaac cagaagtggc
     5161 agcggccata cttcctcctg ttggagcgtt tcatgctttg ctgttgaagt tcgtaactgc
     5221 tgcagcttcg agaactgccc gtggggtgga caaagttttc tgtctctgcc tctgtcccag
     5281 aggcttgcag gccagcaagc tctttggtac gcttttcagt ttccttatag atcctccagt
     5341 ataaaatagt cataatggtg acaggcatat aaaaaccagc gatggctgtg ccaaaagtaa
     5401 tggtgggctc actgaggaac tgaatgaagc actctcccgg aggcacagtt ctctttccaa
     5461 caaagtattg ccagaacaag atggcaggag cccaaaggac aaaggagatg acccaagcca
     5521 gaccgatcat cacaccggct ctctttgttg ttcgtttggc tcggtacgtg agcggcctcg
     5581 tgatggaaaa gtatctgtca aagctgatga ccagaagatt cataacagag gcattgctgg
     5641 ctacgcagtc aatggcaagc cagaggtcac aggccaagtt ccctaaggcc catcgattca
     5701 tgatgatgta ggtcgtaaac agattcattg aaatgacccc gataatcaga tcggcacagg
     5761 ccaggcttaa gaggaagtag ttgttgaccg tcttcagctg cttgttgacc ttaaatgaca
     5821 caattaccag gatgttgccg atgatggtca ccaaggccag gatgcccgtt aagaaagcga
     5881 tgaagaccac ttgccagacg gtatgacctc ccagagggtc atcggtggta ccgtctggag
     5941 aggagaaatt gccagctgct cgagaaacat tgtagctgcc gaaatgagtg acggttcccg
     6001 ggggcagccc tgcatcggag gggctgtgta tccaggagga gctgatgttt ggaaacaaag
     6061 gcgaggttgt actgttattg tgcaaggtgg ggccggggtt ctcctccacg tcgccgcagg
     6121 tcagcaggct gcccctgccc tcatacagac gctggggctt gcccatggtg ctcttgatgt
     6181 acagagcccg cacgttttgc cagtttttct taagcaagga caccaagaaa ttgacagcca
     6241 gatgaatgtt gtagactagc tcatcatcgg tcatcttcac gtggccaaca gcgacggcca
     6301 aacacagcac cttcttcatc tggaacttga ttgtcgattt cacctcatcc actttggcca
     6361 ccatgttttc attgtgtgtc agcagggagg ggaacttgcc agccttgttt aggcctgggc
     6421 ccaggatacg tgggatctgc ttaatcagag actcagaggc caaaaaggca tcgtacttct
     6481 tagccagctt cttgaccaac ttcttgtttt tgttaagctt cttgagcgcc tcgatgtcca
     6541 tgtgggggat atccacggcc ttggcttcat cacagtgctg ctggtccccc agaacgcaca
     6601 ccgagaactt ggggcgtggg gtggacttga gcctgacggt gcccgagaaa cgtttgtcct
     6661 tctgagggtc gtagttcttc aggctgatct gcagctccac cgtctccaga aacttgcggc
     6721 gcttgcgctg gttcccgtgc aggacttccc gcaccgcctc gtacagggtg tcgcgtgaga
     6781 ctttgctgct ggcgtaatct ggaacatcgt atgggtacat ggtggcgcta gcacgcgccg
     6841 aagttcctat actttctaga gaataggaac ttcggaatag gaacttccat tgggattctt
     6901 cctattttga tccaagcatc accatcgacc ctctagtcca gatctcacca tcgacccgaa
     6961 gttcctatac cttttgaaga ataggaactt cggaatagga acttcaagct tatatgcggc
     7021 cgatcataac ttcgtatagc atacattata cgaagttatg aattcgatat caagcttatc
     7081 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     7141 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     7201 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     7261 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     7321 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     7381 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     7441 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     7501 ctcgcctatg ttgccacctg gattctgcgc ggga
//
