LOCUS       sequence-433248-        7379 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the hM3D(Gq) excitatory DREADD fused to mCherry, in a 
            Cre-ON and FLP-OFF dependent manner, under the control of hSyn 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7379)
  TITLE     Somatostatin interneurons control the timing of developmental 
            desynchronization in cortical networks
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7379)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-hSyn-frt-DIO-hM3D(Gq)-mCherry
FEATURES             Location/Qualifiers
     source          1..7379
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    644..677
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(690..723)
                     /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(774..807)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(816..1526)
                     /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(1094..1113)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1363..1381
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1523..1532
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3330..3363
                     /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    3414..3447
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3460..3493
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3506..4094
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3559..3579)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3977..3988)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4126..4602
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4325..4344)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4642..4782
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4642..4771
                     /label=AAV2 ITR
     rep_origin      4857..5312
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4944..4963)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5154..5175
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5329..5348)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5448..5470
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5594..5698
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5699..5767,5768..6559)
                     /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(5917..5936)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6730..7318
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7219..7238
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccttaat taagaagttc ctattctcta
      661 gaaagtatag gaacttcgtc gactccggaa taacttcgta taggatactt tatacgaagt
      721 tatgcagaat ggtagctgga ttgtagctgc tattagcaat atgaaacctc ttaataactt
      781 cgtatagcat acattatacg aagttatggc gcgccttact tgtacagctc gtccatgccg
      841 ccggtggagt ggcggccctc ggcgcgttcg tactgttcca cgatggtgta gtcctcgttg
      901 tgggaggtga tgtccaactt gatgttgacg ttgtaggcgc cgggcagctg cacgggcttc
      961 ttggccttgt aggtggtctt gacctcagcg tcgtagtggc cgccgtcctt cagcttcagc
     1021 ctctgcttga tctcgccctt cagggcgccg tcctcggggt acatccgctc ggaggaggcc
     1081 tcccagccca tggtcttctt ctgcattacg gggccgtcgg aggggaagtt ggtgccgcgc
     1141 agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg agtcctgggt cacggtcacc
     1201 acgccgccgt cctcgaagtt catcacgcgc tcccacttga agccctcggg gaaggacagc
     1261 ttcaagtagt cggggatgtc ggcggggtgc ttcacgtagg ccttggagcc gtacatgaac
     1321 tgaggggaca ggatgtccca ggcgaagggc agggggccac ccttggtcac cttcagcttg
     1381 gcggtctggg tgccctcgta ggggcggccc tcgccctcgc cctcgatctc gaactcgtgg
     1441 ccgttcacgg agccctccat gtgcaccttg aagcgcatga actccttgat gatggccatg
     1501 ttatcctcct cgcccttgct caccatggtg gcgaccgggg gatccttcaa ggcctgctcg
     1561 ggtgcgcgct tgtgaaaaat gaccgactgt ctctgctggt actgctgctt gcgcctcttt
     1621 tttttgtcac actggcacag cagcagcatc ttgaaagtgg ttctgaatgt tttgttgcac
     1681 agagcatagc acacggggtt cacggtgctg ttgatgtagc acagccagta gcccagattc
     1741 caaaaggttt tgggtatgca gctgtcacaa aaggtgttca ccagaaccat gatgttgtat
     1801 ggggtccaag tgatgatgaa ggcaagcaag atcgcactga gggtctgggc cgctttcttc
     1861 tccttgacca gggacatcct tttccgctta gtgatctgac ttctggtctt cagagcaaac
     1921 ctcttggcca gagtggcttc cttgaaggac agaggtagag tggccgtgct cttacccact
     1981 gaggagttga cgtcagaagt cttagctgtg tccacggctg actctagctg gatgggaagc
     2041 ttggagaagc tttttggaaa actgcctcca tcgtccacgc tcttctgggc ctgcagcttg
     2101 tcggctttcc tctccaagtc caccatcccc agctcctcct caggcacctg caggttgtcc
     2161 gatgagggta acttggtgga gttgaggatg gtgctgtgac ccggaagctt gagcacgatg
     2221 gagtagatgg ctctcgtctc ggagccaatg tcctcctcgt cggaggaggc ggagttctcc
     2281 agggaggcag cagcatcatt gttgttccaa ctgtcactgc tgctgtggtc ttggtccatc
     2341 tgctcggagc tgggtttcca gctcttggtt gtgaaccaga agtggcagcg gccatacttc
     2401 ctcctgttgg agcgtttcat gctttgctgt tgaagttcgt aactgctgca gcttcgagaa
     2461 ctgcccgtgg ggtggacaaa gttttctgtc tctgcctctg tcccagaggc ttgcaggcca
     2521 gcaagctctt tggtacgctt ttcagtttcc ttatagatcc tccagtataa aatagtcata
     2581 atggtgacag gcatataaaa accagcgatg gctgtgccaa aagtaatggt gggctcactg
     2641 aggaactgaa tgaagcactc tcccggaggc acagttctct ttccaacaaa gtattgccag
     2701 aacaagatgg caggagccca aaggacaaag gagatgaccc aagccagacc gatcatcaca
     2761 ccggctctct ttgttgttcg tttggctcgg tacgtgagcg gcctcgtgat ggaaaagtat
     2821 ctgtcaaagc tgatgaccag aagattcata acagaggcat tgctggctac gcagtcaatg
     2881 gcaagccaga ggtcacaggc caagttccct aaggcccatc gattcatgat gatgtaggtc
     2941 gtaaacagat tcattgaaat gaccccgata atcagatcgg cacaggccag gcttaagagg
     3001 aagtagttgt tgaccgtctt cagctgcttg ttgaccttaa atgacacaat taccaggatg
     3061 ttgccgatga tggtcaccaa ggccaggatg cccgttaaga aagcgatgaa gaccacttgc
     3121 cagacggtat gacctcccag agggtcatcg gtggtaccgt ctggagagga gaaattgcca
     3181 gctgctcgag aaacattgta gctgccgaaa tgagtgacgg ttcccggggg cagccctgca
     3241 tcggaggggc tgtgtatcca ggaggagctg atgtttggaa acaaaggcga ggttgtactg
     3301 ttattgtgca aggtcatggt ggcgctagca taacttcgta taaagtatcc tatacgaagt
     3361 tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt
     3421 cgtataatgt atgctatacg aagttatgaa ttccctaggg aagttcctat tctctagaaa
     3481 gtataggaac ttcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt
     3541 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     3601 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     3661 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     3721 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     3781 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     3841 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     3901 atcatcgtcc tttccttggc tgctcgccta tgttgccacc tggattctgc gcgggacgtc
     3961 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     4021 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     4081 ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg gcatccctgt
     4141 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     4201 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     4261 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     4321 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     4381 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     4441 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     4501 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     4561 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     4621 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     4681 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     4741 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     4801 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     4861 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     4921 acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     4981 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     5041 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     5101 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     5161 cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg atttataagg
     5221 gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     5281 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     5341 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     5401 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5461 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     5521 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     5581 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     5641 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     5701 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     5761 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     5821 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5881 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5941 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     6001 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     6061 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     6121 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     6181 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     6241 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     6301 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     6361 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     6421 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     6481 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     6541 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     6601 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     6661 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     6721 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     6781 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     6841 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     6901 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6961 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     7021 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     7081 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     7141 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     7201 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     7261 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     7321 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
