LOCUS       sequence-444377-        7404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses hM3D(Gq) and a control shRNA (targetting LacZ) 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-shLacZ-CWB
FEATURES             Location/Qualifiers
     source          1..7404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1023..1056
                     /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)."
     protein_bind    1107..1140
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1176..1195
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    1333..1921
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1386..1406)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1804..1815)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    1934..2158
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2166..2306
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2166..2295
                     /label=AAV2 ITR
     rep_origin      2381..2836
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2468..2487)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2678..2699
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2853..2872)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2972..2994
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        3118..3222
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3223..3291,3292..4083)
                     /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(3441..3460)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4254..4842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4743..4762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   4904..5033
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        5078..5525
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(5590..5623)
                     /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)."
     protein_bind    complement(5674..5707)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(5796..5816)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(5908..6618)
                     /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(6186..6205)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6455..6473
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      6615..6624
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 gtcactgctg ctgtggtctt ggtccatctg ctcggagctg ggtttccagc tcttggttgt
       61 gaaccagaag tggcagcggc catacttcct cctgttggag cgtttcatgc tttgctgttg
      121 aagttcgtaa ctgctgcagc ttcgagaact gcccgtgggg tggacaaagt tttctgtctc
      181 tgcctctgtc ccagaggctt gcaggccagc aagctctttg gtacgctttt cagtttcctt
      241 atagatcctc cagtataaaa tagtcataat ggtgacaggc atataaaaac cagcgatggc
      301 tgtgccaaaa gtaatggtgg gctcactgag gaactgaatg aagcactctc ccggaggcac
      361 agttctcttt ccaacaaagt attgccagaa caagatggca ggagcccaaa ggacaaagga
      421 gatgacccaa gccagaccga tcatcacacc ggctctcttt gttgttcgtt tggctcggta
      481 cgtgagcggc ctcgtgatgg aaaagtatct gtcaaagctg atgaccagaa gattcataac
      541 agaggcattg ctggctacgc agtcaatggc aagccagagg tcacaggcca agttccctaa
      601 ggcccatcga ttcatgatga tgtaggtcgt aaacagattc attgaaatga ccccgataat
      661 cagatcggca caggccaggc ttaagaggaa gtagttgttg accgtcttca gctgcttgtt
      721 gaccttaaat gacacaatta ccaggatgtt gccgatgatg gtcaccaagg ccaggatgcc
      781 cgttaagaaa gcgatgaaga ccacttgcca gacggtatga cctcccagag ggtcatcggt
      841 ggtaccgtct ggagaggaga aattgccagc tgctcgagaa acattgtagc tgccgaaatg
      901 agtgacggtt cccgggggca gccctgcatc ggaggggctg tgtatccagg aggagctgat
      961 gtttggaaac aaaggcgagg ttgtactgtt attgtgcaag gtcatggtgg cggcgcgccg
     1021 cgataacttc gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac
     1081 tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat
     1141 gtttatctgc agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt
     1201 gaaagtattt cgatttcttg ggtttatata tcttgtggaa aggacgcggt ttcctagaaa
     1261 tcgctgattt gtgtagtccc tgacccagac tacacaaatc agcgattttt tttgaattcc
     1321 tgcagccccg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     1381 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     1441 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     1501 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     1561 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     1621 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     1681 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     1741 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     1801 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     1861 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     1921 catcgggggg atcctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     1981 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2041 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2101 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggcg
     2161 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     2221 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     2281 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     2341 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     2401 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     2461 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     2521 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     2581 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     2641 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     2701 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     2761 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     2821 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     2881 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     2941 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3001 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     3061 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3121 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3181 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3241 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3301 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3361 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3421 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3481 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3541 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3601 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3661 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3721 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3781 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3841 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3901 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3961 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4021 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4081 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     4141 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4201 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4261 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4321 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4381 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4621 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     4921 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     4981 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     5041 cacgcgtgtg tctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggacca
     5101 ggatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg gacaagcacc
     5161 caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga aacaggatgc
     5221 ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg cccccgcctg
     5281 gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc gccggtcccc
     5341 cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc ccagccggac
     5401 cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc tgcggcgccg
     5461 gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg tgcctgagag
     5521 cgcagtcgag aaggtaccgg atcctctaga gtcgactccg gagtcgactc tagagctagc
     5581 ggatccggaa taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga
     5641 ttgtagctgc tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg
     5701 aagttattat tttgtactaa tatctttgtg tttacagtca aattaattct aattatctct
     5761 ctaacagcct tgtatcgtat atgcaaatat gaaggaatca tgggaaatag gccctcttcc
     5821 tgcccgacct tggcgcgcgc tcggcgcgcg gtcacgctcc gtcacgtggt gcgttttgcc
     5881 tgcgcgtctt tccactgggt taattaatta cttgtacagc tcgtccatgc cgccggtgga
     5941 gtggcggccc tcggcgcgtt cgtactgttc cacgatggtg tagtcctcgt tgtgggaggt
     6001 gatgtccaac ttgatgttga cgttgtaggc gccgggcagc tgcacgggct tcttggcctt
     6061 gtaggtggtc ttgacctcag cgtcgtagtg gccgccgtcc ttcagcttca gcctctgctt
     6121 gatctcgccc ttcagggcgc cgtcctcggg gtacatccgc tcggaggagg cctcccagcc
     6181 catggtcttc ttctgcatta cggggccgtc ggaggggaag ttggtgccgc gcagcttcac
     6241 cttgtagatg aactcgccgt cctgcaggga ggagtcctgg gtcacggtca ccacgccgcc
     6301 gtcctcgaag ttcatcacgc gctcccactt gaagccctcg gggaaggaca gcttcaagta
     6361 gtcggggatg tcggcggggt gcttcacgta ggccttggag ccgtacatga actgagggga
     6421 caggatgtcc caggcgaagg gcagggggcc acccttggtc accttcagct tggcggtctg
     6481 ggtgccctcg taggggcggc cctcgccctc gccctcgatc tcgaactcgt ggccgttcac
     6541 ggagccctcc atgtgcacct tgaagcgcat gaactccttg atgatggcca tgttatcctc
     6601 ctcgcccttg ctcaccatgg tggcgaccgg gggatccttc aaggcctgct cgggtgcgcg
     6661 cttgtgaaaa atgaccgact gtctctgctg gtactgctgc ttgcgcctct tttttttgtc
     6721 acactggcac agcagcagca tcttgaaagt ggttctgaat gttttgttgc acagagcata
     6781 gcacacgggg ttcacggtgc tgttgatgta gcacagccag tagcccagat tccaaaaggt
     6841 tttgggtatg cagctgtcac aaaaggtgtt caccagaacc atgatgttgt atggggtcca
     6901 agtgatgatg aaggcaagca agatcgcact gagggtctgg gccgctttct tctccttgac
     6961 cagggacatc cttttccgct tagtgatctg acttctggtc ttcagagcaa acctcttggc
     7021 cagagtggct tccttgaagg acagaggtag agtggccgtg ctcttaccca ctgaggagtt
     7081 gacgtcagaa gtcttagctg tgtccacggc tgactctagc tggatgggaa gcttggagaa
     7141 gctttttgga aaactgcctc catcgtccac gctcttctgg gcctgcagct tgtcggcttt
     7201 cctctccaag tccaccatcc ccagctcctc ctcaggcacc tgcaggttgt ccgatgaggg
     7261 taacttggtg gagttgagga tggtgctgtg acccggaagc ttgagcacga tggagtagat
     7321 ggctctcgtc tcggagccaa tgtcctcctc gtcggaggag gcggagttct ccagggaggc
     7381 agcagcatca ttgttgttcc aact
//
