LOCUS       sequence-444385-        7404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hM3D(Gq) and a shRNA targetting VGF in a Cre-dependent 
            manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7404)
  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 7404)
  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-DIO-hM3D(Gq)-mCherry-shVgf-CWB
FEATURES             Location/Qualifiers
     source          1..7404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(90..109)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(145..178)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(229..262)
                     /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)."
     regulatory      2065..2074
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2071..2781
                     /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(2216..2234)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2484..2503
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2873..2893
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     protein_bind    2982..3015
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3066..3099
                     /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)."
     promoter        complement(3164..3611)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   3656..3785
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3847..4435)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3927..3946)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4606..5397,5398..5466))
                     /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     5229..5248
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5467..5571)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5695..5717)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5817..5836
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5853..6308)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5990..6011)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6202..6221
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   6383..6523
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   6394..6523
                     /label=AAV2 ITR
     polyA_signal    6531..6755
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    6768..7356
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6874..6885
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     7283..7303
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 gtcaggtcaa tgagactatc gatcgtccta ggaaaccgcg tcctttccac aagatatata
       61 aacccaagaa atcgaaatac tttcaagtta cggtaagcat atgatagtcc attttaaaac
      121 ataattttaa aactgcagat aaacataact tcgtatagca tacattatac gaagttattc
      181 tttgcaccat tctaaagaat aacagtgata atttctgggt taaggcaaat aacttcgtat
      241 aggatacttt atacgaagtt atcgcggcgc gccgccacca tgaccttgca caataacagt
      301 acaacctcgc ctttgtttcc aaacatcagc tcctcctgga tacacagccc ctccgatgca
      361 gggctgcccc cgggaaccgt cactcatttc ggcagctaca atgtttctcg agcagctggc
      421 aatttctcct ctccagacgg taccaccgat gaccctctgg gaggtcatac cgtctggcaa
      481 gtggtcttca tcgctttctt aacgggcatc ctggccttgg tgaccatcat cggcaacatc
      541 ctggtaattg tgtcatttaa ggtcaacaag cagctgaaga cggtcaacaa ctacttcctc
      601 ttaagcctgg cctgtgccga tctgattatc ggggtcattt caatgaatct gtttacgacc
      661 tacatcatca tgaatcgatg ggccttaggg aacttggcct gtgacctctg gcttgccatt
      721 gactgcgtag ccagcaatgc ctctgttatg aatcttctgg tcatcagctt tgacagatac
      781 ttttccatca cgaggccgct cacgtaccga gccaaacgaa caacaaagag agccggtgtg
      841 atgatcggtc tggcttgggt catctccttt gtcctttggg ctcctgccat cttgttctgg
      901 caatactttg ttggaaagag aactgtgcct ccgggagagt gcttcattca gttcctcagt
      961 gagcccacca ttacttttgg cacagccatc gctggttttt atatgcctgt caccattatg
     1021 actattttat actggaggat ctataaggaa actgaaaagc gtaccaaaga gcttgctggc
     1081 ctgcaagcct ctgggacaga ggcagagaca gaaaactttg tccaccccac gggcagttct
     1141 cgaagctgca gcagttacga acttcaacag caaagcatga aacgctccaa caggaggaag
     1201 tatggccgct gccacttctg gttcacaacc aagagctgga aacccagctc cgagcagatg
     1261 gaccaagacc acagcagcag tgacagttgg aacaacaatg atgctgctgc ctccctggag
     1321 aactccgcct cctccgacga ggaggacatt ggctccgaga cgagagccat ctactccatc
     1381 gtgctcaagc ttccgggtca cagcaccatc ctcaactcca ccaagttacc ctcatcggac
     1441 aacctgcagg tgcctgagga ggagctgggg atggtggact tggagaggaa agccgacaag
     1501 ctgcaggccc agaagagcgt ggacgatgga ggcagttttc caaaaagctt ctccaagctt
     1561 cccatccagc tagagtcagc cgtggacaca gctaagactt ctgacgtcaa ctcctcagtg
     1621 ggtaagagca cggccactct acctctgtcc ttcaaggaag ccactctggc caagaggttt
     1681 gctctgaaga ccagaagtca gatcactaag cggaaaagga tgtccctggt caaggagaag
     1741 aaagcggccc agaccctcag tgcgatcttg cttgccttca tcatcacttg gaccccatac
     1801 aacatcatgg ttctggtgaa caccttttgt gacagctgca tacccaaaac cttttggaat
     1861 ctgggctact ggctgtgcta catcaacagc accgtgaacc ccgtgtgcta tgctctgtgc
     1921 aacaaaacat tcagaaccac tttcaagatg ctgctgctgt gccagtgtga caaaaaaaag
     1981 aggcgcaagc agcagtacca gcagagacag tcggtcattt ttcacaagcg cgcacccgag
     2041 caggccttga aggatccccc ggtcgccacc atggtgagca agggcgagga ggataacatg
     2101 gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt gaacggccac
     2161 gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca gaccgccaag
     2221 ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc ccctcagttc
     2281 atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttgaagctg
     2341 tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg cggcgtggtg
     2401 accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt gaagctgcgc
     2461 ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg ctgggaggcc
     2521 tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa gcagaggctg
     2581 aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa ggccaagaag
     2641 cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac ctcccacaac
     2701 gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc
     2761 atggacgagc tgtacaagta attaattaac ccagtggaaa gacgcgcagg caaaacgcac
     2821 cacgtgacgg agcgtgaccg cgcgccgagc gcgcgccaag gtcgggcagg aagagggcct
     2881 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattag
     2941 aattaatttg actgtaaaca caaagatatt agtacaaaat aataacttcg tataatgtat
     3001 gctatacgaa gttattaaga ggtttcatat tgctaatagc agctacaatc cagctaccat
     3061 tctgcataac ttcgtataaa gtatcctata cgaagttatt ccggatccgc tagctctaga
     3121 gtcgactccg gagtcgactc tagaggatcc ggtaccttct cgactgcgct ctcaggcacg
     3181 acacgactcc tccgctgccc accgcagact gaggcagcgc tgagtcgccg gcgccgcagc
     3241 gcagatggtc gcgcccgtgc ccccctatct cgcgcctcgc gtggtgcggt ccggctgggc
     3301 cggcggcggc gcggacgcga ccaaggtggc cgggaagggg agtttgcggg ggaccggcga
     3361 gtgacgtcag cgcgccttca gtgctgaggc ggcggtggcg cgcgccgcca ggcgggggcg
     3421 aaggcactgt ccgcggtgct gaagctggca gtgcgcacgc gcctcgccgc atcctgtttc
     3481 ccctccccct ctctgatagg ggatgcgcaa tttggggaat gggggttggg tgcttgtcca
     3541 gtgggtcggg gtcggtcgtc aggtaggcac ccccaccccg cctcatcctg gtcctaaaac
     3601 ccacttgcac tcatacgcag ggccctctgc agtctagaca cacgcgtgcg gccgcaggaa
     3661 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg
     3721 cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     3781 gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3841 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3901 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3961 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     4021 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     4081 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     4141 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     4201 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4261 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4321 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4381 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4441 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4501 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4561 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4621 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4681 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4741 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4801 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4861 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4921 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4981 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     5041 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     5101 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     5161 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     5221 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5281 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5341 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5401 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5461 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5521 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5581 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     5641 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     5701 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     5761 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     5821 atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata ttttgttaaa
     5881 attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg aaatcggcaa
     5941 aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc cagtttggaa
     6001 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     6061 gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt cgaggtgccg
     6121 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     6181 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta aggcgctggc
     6241 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     6301 gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag atgcgtaagg
     6361 agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct cactgaggcc
     6421 gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga
     6481 gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg ccatagagcc
     6541 caccgcatcc ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc
     6601 caccccaccc cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt
     6661 ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg
     6721 gcaaacaaca gatggctggc aactagaagg cacaggatcc ccccgatgcg gggaggcggc
     6781 ccaaagggag atccgactcg tctgagggcg aaggcgaaga cgcggaagag gccgcagagc
     6841 cggcagcagg ccgcgggaag gaaggtccgc tggattgagg gccgaaggga cgtagcagaa
     6901 ggacgtcccg cgcagaatcc aggtggcaac acaggcgagc agccaaggaa aggacgatga
     6961 tttccccgac aacaccacgg aattgtcagt gcccaacagc cgagcccctg tccagcagcg
     7021 ggcaaggcag gcggcgatga gttccgccgt ggcaataggg agggggaaag cgaaagtccc
     7081 ggaaaggagc tgacaggtgg tggcaatgcc ccaaccagtg ggggttgcgt cagcaaacac
     7141 agtgcacacc acgccacgtt gcctgacaac gggccacaac tcctcataaa gagacagcaa
     7201 ccaggattta tacaaggagg agaaaatgaa agccatacgg gaagcaatag catgatacaa
     7261 aggcattaaa gcagcgtatc cacatagcgt aaaaggagca acatagttaa gaataccagt
     7321 caatctttca caaattttgt aatccagagg ttgattatcg gggctgcagg aattcaaaaa
     7381 gacgatcgat agtctcattg atgg
//
