LOCUS       sequence-444380-        7402 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hM3D(Gq) and a shRNA targetting Scg2 in a Cre-dependent 
            manner.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7402)
  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 7402)
  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-shScg2-CWB
FEATURES             Location/Qualifiers
     source          1..7402
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(123..141)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     391..410
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     780..800
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     protein_bind    889..922
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    973..1006
                     /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(1071..1518)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   1563..1692
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(1754..2342)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1834..1853)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2513..3304,3305..3373))
                     /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     3136..3155
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3374..3478)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3602..3624)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3724..3743
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3760..4215)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3897..3918)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4109..4128
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4290..4430
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4301..4430
                     /label=AAV2 ITR
     polyA_signal    4438..4662
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    4675..5263
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             4781..4792
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     5190..5210
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     primer_bind     complement(5399..5418)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(5454..5487)
                     /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(5538..5571)
                     /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      7374..7383
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7380..7402,1..688)
                     /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"
ORIGIN
        1 taacatggcc atcatcaagg agttcatgcg cttcaaggtg cacatggagg gctccgtgaa
       61 cggccacgag ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac
      121 cgccaagctg aaggtgacca agggtggccc cctgcccttc gcctgggaca tcctgtcccc
      181 tcagttcatg tacggctcca aggcctacgt gaagcacccc gccgacatcc ccgactactt
      241 gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg
      301 cgtggtgacc gtgacccagg actcctccct gcaggacggc gagttcatct acaaggtgaa
      361 gctgcgcggc accaacttcc cctccgacgg ccccgtaatg cagaagaaga ccatgggctg
      421 ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
      481 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
      541 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atcaagttgg acatcacctc
      601 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
      661 cggcggcatg gacgagctgt acaagtaatt aattaaccca gtggaaagac gcgcaggcaa
      721 aacgcaccac gtgacggagc gtgaccgcgc gccgagcgcg cgccaaggtc gggcaggaag
      781 agggcctatt tcccatgatt ccttcatatt tgcatatacg atacaaggct gttagagaga
      841 taattagaat taatttgact gtaaacacaa agatattagt acaaaataat aacttcgtat
      901 aatgtatgct atacgaagtt attaagaggt ttcatattgc taatagcagc tacaatccag
      961 ctaccattct gcataacttc gtataaagta tcctatacga agttattccg gatccgctag
     1021 ctctagagtc gactccggag tcgactctag aggatccggt accttctcga ctgcgctctc
     1081 aggcacgaca cgactcctcc gctgcccacc gcagactgag gcagcgctga gtcgccggcg
     1141 ccgcagcgca gatggtcgcg cccgtgcccc cctatctcgc gcctcgcgtg gtgcggtccg
     1201 gctgggccgg cggcggcgcg gacgcgacca aggtggccgg gaaggggagt ttgcggggga
     1261 ccggcgagtg acgtcagcgc gccttcagtg ctgaggcggc ggtggcgcgc gccgccaggc
     1321 gggggcgaag gcactgtccg cggtgctgaa gctggcagtg cgcacgcgcc tcgccgcatc
     1381 ctgtttcccc tccccctctc tgatagggga tgcgcaattt ggggaatggg ggttgggtgc
     1441 ttgtccagtg ggtcggggtc ggtcgtcagg taggcacccc caccccgcct catcctggtc
     1501 ctaaaaccca cttgcactca tacgcagggc cctctgcagt ctagacacac gcgtgcggcc
     1561 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
     1621 ggccgggcga ccaaaggtcg cccgacgccc gggcggcctc agtgagcgag cgagcgcgca
     1681 gctgcctgca ggacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     1741 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     1801 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     1861 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     1921 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1981 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     2041 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     2101 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     2161 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     2221 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2281 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2341 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     2401 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     2461 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     2521 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     2581 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     2641 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     2701 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     2761 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     2821 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     2881 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     2941 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     3001 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     3061 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     3121 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     3181 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     3241 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     3301 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     3361 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     3421 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     3481 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     3541 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     3601 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     3661 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     3721 atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac gttaatattt
     3781 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa tagaccgaaa
     3841 tcggcaaaat cccttataaa tcaaaagaat agcccgagat agagttgagt gttgttccag
     3901 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3961 tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt ttggggtcga
     4021 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     4081 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaagg
     4141 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     4201 cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac cgcacagatg
     4261 cgtaaggaga aaataccgca tcaggcgccc ctgcaggcag ctgcgcgctc gctcgctcac
     4321 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
     4381 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcca
     4441 tagagcccac cgcatcccca gcatgcctgc tattgtcttc ccaatcctcc cccttgctgt
     4501 cctgccccac cccacccccc agaatagaat gacacctact cagacaatgc gatgcaattt
     4561 cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag gaaggcacgg
     4621 gggaggggca aacaacagat ggctggcaac tagaaggcac aggatccccc cgatgcgggg
     4681 aggcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc ggaagaggcc
     4741 gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc gaagggacgt
     4801 agcagaagga cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc caaggaaagg
     4861 acgatgattt ccccgacaac accacggaat tgtcagtgcc caacagccga gcccctgtcc
     4921 agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg gggaaagcga
     4981 aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg gttgcgtcag
     5041 caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc tcataaagag
     5101 acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
     5161 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
     5221 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatcgggg ctgcaggaat
     5281 tcaaaaagca gacaagcacc ttatgaatgg gtcaggttca taaggtgctt gtctgcctag
     5341 gaaaccgcgt cctttccaca agatatataa acccaagaaa tcgaaatact ttcaagttac
     5401 ggtaagcata tgatagtcca ttttaaaaca taattttaaa actgcagata aacataactt
     5461 cgtatagcat acattatacg aagttattct ttgcaccatt ctaaagaata acagtgataa
     5521 tttctgggtt aaggcaaata acttcgtata ggatacttta tacgaagtta tcgcggcgcg
     5581 ccgccaccat gaccttgcac aataacagta caacctcgcc tttgtttcca aacatcagct
     5641 cctcctggat acacagcccc tccgatgcag ggctgccccc gggaaccgtc actcatttcg
     5701 gcagctacaa tgtttctcga gcagctggca atttctcctc tccagacggt accaccgatg
     5761 accctctggg aggtcatacc gtctggcaag tggtcttcat cgctttctta acgggcatcc
     5821 tggccttggt gaccatcatc ggcaacatcc tggtaattgt gtcatttaag gtcaacaagc
     5881 agctgaagac ggtcaacaac tacttcctct taagcctggc ctgtgccgat ctgattatcg
     5941 gggtcatttc aatgaatctg tttacgacct acatcatcat gaatcgatgg gccttaggga
     6001 acttggcctg tgacctctgg cttgccattg actgcgtagc cagcaatgcc tctgttatga
     6061 atcttctggt catcagcttt gacagatact tttccatcac gaggccgctc acgtaccgag
     6121 ccaaacgaac aacaaagaga gccggtgtga tgatcggtct ggcttgggtc atctcctttg
     6181 tcctttgggc tcctgccatc ttgttctggc aatactttgt tggaaagaga actgtgcctc
     6241 cgggagagtg cttcattcag ttcctcagtg agcccaccat tacttttggc acagccatcg
     6301 ctggttttta tatgcctgtc accattatga ctattttata ctggaggatc tataaggaaa
     6361 ctgaaaagcg taccaaagag cttgctggcc tgcaagcctc tgggacagag gcagagacag
     6421 aaaactttgt ccaccccacg ggcagttctc gaagctgcag cagttacgaa cttcaacagc
     6481 aaagcatgaa acgctccaac aggaggaagt atggccgctg ccacttctgg ttcacaacca
     6541 agagctggaa acccagctcc gagcagatgg accaagacca cagcagcagt gacagttgga
     6601 acaacaatga tgctgctgcc tccctggaga actccgcctc ctccgacgag gaggacattg
     6661 gctccgagac gagagccatc tactccatcg tgctcaagct tccgggtcac agcaccatcc
     6721 tcaactccac caagttaccc tcatcggaca acctgcaggt gcctgaggag gagctgggga
     6781 tggtggactt ggagaggaaa gccgacaagc tgcaggccca gaagagcgtg gacgatggag
     6841 gcagttttcc aaaaagcttc tccaagcttc ccatccagct agagtcagcc gtggacacag
     6901 ctaagacttc tgacgtcaac tcctcagtgg gtaagagcac ggccactcta cctctgtcct
     6961 tcaaggaagc cactctggcc aagaggtttg ctctgaagac cagaagtcag atcactaagc
     7021 ggaaaaggat gtccctggtc aaggagaaga aagcggccca gaccctcagt gcgatcttgc
     7081 ttgccttcat catcacttgg accccataca acatcatggt tctggtgaac accttttgtg
     7141 acagctgcat acccaaaacc ttttggaatc tgggctactg gctgtgctac atcaacagca
     7201 ccgtgaaccc cgtgtgctat gctctgtgca acaaaacatt cagaaccact ttcaagatgc
     7261 tgctgctgtg ccagtgtgac aaaaaaaaga ggcgcaagca gcagtaccag cagagacagt
     7321 cggtcatttt tcacaagcgc gcacccgagc aggccttgaa ggatcccccg gtcgccacca
     7381 tggtgagcaa gggcgaggag ga
//
