LOCUS       sequence-444382-        7478 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hM3D(Gq) in a flippase dependent manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7478)
  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 7478)
  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-fDIO-hM3D(Gq)-mCherry
FEATURES             Location/Qualifiers
     source          1..7478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(1663..1710)
                     /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)."
     misc_feature    1876..2464
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1929..1949)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2347..2358)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2496..2972
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2695..2714)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3012..3152
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3012..3141
                     /label=AAV2 ITR
     rep_origin      3227..3682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3314..3333)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3524..3545
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3699..3718)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3818..3840
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3964..4068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4069..4137,4138..4929)
                     /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(4287..4306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5100..5688
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5589..5608
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5750..5879
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        5924..6371
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    6449..6496
                     /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(6620..7330)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(6898..6917)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7167..7185
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      7327..7336
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 ttgcacagag catagcacac ggggttcacg gtgctgttga tgtagcacag ccagtagccc
       61 agattccaaa aggttttggg tatgcagctg tcacaaaagg tgttcaccag aaccatgatg
      121 ttgtatgggg tccaagtgat gatgaaggca agcaagatcg cactgagggt ctgggccgct
      181 ttcttctcct tgaccaggga catccttttc cgcttagtga tctgacttct ggtcttcaga
      241 gcaaacctct tggccagagt ggcttccttg aaggacagag gtagagtggc cgtgctctta
      301 cccactgagg agttgacgtc agaagtctta gctgtgtcca cggctgactc tagctggatg
      361 ggaagcttgg agaagctttt tggaaaactg cctccatcgt ccacgctctt ctgggcctgc
      421 agcttgtcgg ctttcctctc caagtccacc atccccagct cctcctcagg cacctgcagg
      481 ttgtccgatg agggtaactt ggtggagttg aggatggtgc tgtgacccgg aagcttgagc
      541 acgatggagt agatggctct cgtctcggag ccaatgtcct cctcgtcgga ggaggcggag
      601 ttctccaggg aggcagcagc atcattgttg ttccaactgt cactgctgct gtggtcttgg
      661 tccatctgct cggagctggg tttccagctc ttggttgtga accagaagtg gcagcggcca
      721 tacttcctcc tgttggagcg tttcatgctt tgctgttgaa gttcgtaact gctgcagctt
      781 cgagaactgc ccgtggggtg gacaaagttt tctgtctctg cctctgtccc agaggcttgc
      841 aggccagcaa gctctttggt acgcttttca gtttccttat agatcctcca gtataaaata
      901 gtcataatgg tgacaggcat ataaaaacca gcgatggctg tgccaaaagt aatggtgggc
      961 tcactgagga actgaatgaa gcactctccc ggaggcacag ttctctttcc aacaaagtat
     1021 tgccagaaca agatggcagg agcccaaagg acaaaggaga tgacccaagc cagaccgatc
     1081 atcacaccgg ctctctttgt tgttcgtttg gctcggtacg tgagcggcct cgtgatggaa
     1141 aagtatctgt caaagctgat gaccagaaga ttcataacag aggcattgct ggctacgcag
     1201 tcaatggcaa gccagaggtc acaggccaag ttccctaagg cccatcgatt catgatgatg
     1261 taggtcgtaa acagattcat tgaaatgacc ccgataatca gatcggcaca ggccaggctt
     1321 aagaggaagt agttgttgac cgtcttcagc tgcttgttga ccttaaatga cacaattacc
     1381 aggatgttgc cgatgatggt caccaaggcc aggatgcccg ttaagaaagc gatgaagacc
     1441 acttgccaga cggtatgacc tcccagaggg tcatcggtgg taccgtctgg agaggagaaa
     1501 ttgccagctg ctcgagaaac attgtagctg ccgaaatgag tgacggttcc cgggggcagc
     1561 cctgcatcgg aggggctgtg tatccaggag gagctgatgt ttggaaacaa aggcgaggtt
     1621 gtactgttat tgtgcaaggt catggtggcg ctagcacgcg ccgaagttcc tatactttct
     1681 agagaatagg aacttcggaa taggaacttc cattgggatt cttcctattt tgatccaagc
     1741 atcaccatcg accctctagt ccagatctca ccatcgaccc gaagttccta taccttttga
     1801 agaataggaa cttcggaata ggaacttcaa gcttatatgc ggccgatcga tgaattcgat
     1861 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     1921 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     1981 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2041 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2101 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2161 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     2221 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     2281 tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac
     2341 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     2401 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     2461 ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc
     2521 cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa
     2581 aattaagttg catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg
     2641 gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt
     2701 gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg
     2761 ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca
     2821 ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg
     2881 tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac
     2941 aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac
     3001 cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc
     3061 gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc
     3121 agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac
     3181 gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc
     3241 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3301 gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3361 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3421 ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag
     3481 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3541 actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg gattttgccg
     3601 atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     3661 aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca
     3721 tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     3781 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     3841 ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta
     3901 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3961 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     4021 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     4081 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     4141 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     4201 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     4261 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     4321 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     4381 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     4441 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     4501 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4561 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4621 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4681 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4741 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4801 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4861 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4921 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     4981 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5041 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5101 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5161 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5221 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     5281 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5341 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5401 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5461 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5521 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5581 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5641 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5701 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtc ctgcaggcag
     5761 ctgcgcgctc gctcgctcac tgaggccgcc cgggcgtcgg gcgacctttg gtcgcccggc
     5821 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctg
     5881 cggccgcacg cgtgtgtcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
     5941 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
     6001 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
     6061 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
     6121 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
     6181 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
     6241 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
     6301 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
     6361 tgagagcgca gtcgagaagg taccggatcc tgtgctgtct catcattttg gcaaagaatt
     6421 gatttgatac cgcgggcccg ggggtaccga agttcctatt ccgaagttcc tattctctag
     6481 aaagtatagg aacttcatca aaataggaag accaatgctt caccatcgac ccgaattgcc
     6541 aagcatcacc atcgacagaa gttcctattc cgaagttcct attcttcaaa aggtatagga
     6601 acttcgtcga cggcgcgcct tacttgtaca gctcgtccat gccgccggtg gagtggcggc
     6661 cctcggcgcg ttcgtactgt tccacgatgg tgtagtcctc gttgtgggag gtgatgtcca
     6721 acttgatgtt gacgttgtag gcgccgggca gctgcacggg cttcttggcc ttgtaggtgg
     6781 tcttgacctc agcgtcgtag tggccgccgt ccttcagctt cagcctctgc ttgatctcgc
     6841 ccttcagggc gccgtcctcg gggtacatcc gctcggagga ggcctcccag cccatggtct
     6901 tcttctgcat tacggggccg tcggagggga agttggtgcc gcgcagcttc accttgtaga
     6961 tgaactcgcc gtcctgcagg gaggagtcct gggtcacggt caccacgccg ccgtcctcga
     7021 agttcatcac gcgctcccac ttgaagccct cggggaagga cagcttcaag tagtcgggga
     7081 tgtcggcggg gtgcttcacg taggccttgg agccgtacat gaactgaggg gacaggatgt
     7141 cccaggcgaa gggcaggggg ccacccttgg tcaccttcag cttggcggtc tgggtgccct
     7201 cgtaggggcg gccctcgccc tcgccctcga tctcgaactc gtggccgttc acggagccct
     7261 ccatgtgcac cttgaagcgc atgaactcct tgatgatggc catgttatcc tcctcgccct
     7321 tgctcaccat ggtggcgacc gggggatcct tcaaggcctg ctcgggtgcg cgcttgtgaa
     7381 aaatgaccga ctgtctctgc tggtactgct gcttgcgcct cttttttttg tcacactggc
     7441 acagcagcag catcttgaaa gtggttctga atgttttg
//
