LOCUS       sequence-433249-        7046 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the hM4D(Gi) inhibitory 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 7046)
  TITLE     Somatostatin interneurons control the timing of developmental 
            desynchronization in cortical networks
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7046)
  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            "Unpublished"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-hM4D(Gi)-mCherry
FEATURES             Location/Qualifiers
     source          1..7046
                     /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"
     regulatory      2981..2990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2997..3030
                     /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    3081..3114
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    3127..3160
                     /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    3173..3761
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3226..3246)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3644..3655)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3793..4269
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3992..4011)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4309..4449
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4309..4438
                     /label=AAV2 ITR
     rep_origin      4524..4979
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4611..4630)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4821..4842
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4996..5015)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5115..5137
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5175..5193)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5261..5365
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5366..5434,5435..6226)
                     /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(5584..5603)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6397..6985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6886..6905
                     /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 gcgaccggtg gatcccgcct ggcagtgccg
     1561 atgttccgat actggcacag cagcaggtgc cggaaggtct ttttaaaggt ggcgttgcac
     1621 agagcatagc aggcagggtt gatggtgctg ttgacgtagc agagccagta gccaatggac
     1681 cacaccgtgt cagggatgca gctctggcag aaggtgttca ccaggaccat gacgttgtag
     1741 ggcgtccagg tgaggatgaa ggccagcaga atggcaaaga tcgttcgtgt cactttgcgc
     1801 tcccgggccg ccatctgccg cttcttgcgc acctggttgc gagcgatgct ggcgaacttg
     1861 cgggccacgt tggccgcagg gcgcatgcca gccggcgtgg caggcacaat ctcaatggct
     1921 gtcacacact cattgcctgt ctgcttcgtc acaatctgga tcttggacca tctggaggct
     1981 gggttgaggg cccgcggctg caggggaggg gcgggcatgg cgggcgtggt ggcctctgtg
     2041 gtggacagct ctgtggctgg gcgttccttg gtgttctggg tggcactgcc tgagctggac
     2101 tcattggaag tgtccttatc agccacgggg cgcggtggcg gtggcagcgc tggcgggggg
     2161 gcctcctcca gcttgccatt gcgcagctcc tcccgggcgg cctccccggg cgggggcttc
     2221 ttgacgctct gcttcattag tgggctcttg aggaaggcca gcgtcttggc tttcttctcc
     2281 ttcgggccct cgggccggtg cttgtggact cggctgcgac tggccaggga gatgtggatg
     2341 tacagcaccg tcatgatgac cacaggcagg tagaagccag caatggctgt gccaaaggtc
     2401 actgctgggt tggacaggaa ctggatgaag cactggttgt cgggcaccgt ccgcttaccc
     2461 accacaaact gccagaacaa gatggcaggc gcccagagca cgaaggacag tacccaggca
     2521 gcagcaatca tgaggcctgc catcttggtg gtgcgccggg cagggtaggt gagaggcttg
     2581 gtgacgcaga agtagcggtc aaagctgatg atgagaaggt tcatgacgga ggcgttgctc
     2641 accacgcagt ccagggccag ccacaggtcg cagaccacgg cgcccagggg ccagtagccc
     2701 ttgatgatgt acacggtgta gaggttcatg gagaaggcgc ctatgatgag atcagcacac
     2761 gccaggctga agaggaagta gttgttgact gtctgcagct gcctgttgac cttgatggac
     2821 agcatcacca ggatgttgcc cacgacagtc accaggctca gggagcctgt cactgtggca
     2881 atgaagacca tttccaccgt ctcatagcga ttgtgggatg atgacgtgac caggcgcacg
     2941 gactgattgc ccgagctgcc attgacaggt gtgaagttgg ccatggtggc gctagcataa
     3001 cttcgtataa agtatcctat acgaagttat ttgccttaac ccagaaatta tcactgttat
     3061 tctttagaat ggtgcaaaga ataacttcgt ataatgtatg ctatacgaag ttatgaattc
     3121 cctagggaag ttcctattct ctagaaagta taggaacttc aagcttatcg ataatcaacc
     3181 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     3241 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     3301 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     3361 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     3421 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3481 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3541 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3601 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3661 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3721 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     3781 cgagagatct acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa
     3841 gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct
     3901 gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa
     3961 gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg
     4021 gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag
     4081 cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt
     4141 tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg
     4201 atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc
     4261 cctgtccttc tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag
     4321 tgatggagtt ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa
     4381 aggtcgcccg acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct
     4441 gcctgcaggg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     4501 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4561 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
     4621 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4681 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     4741 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4801 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
     4861 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
     4921 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
     4981 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     5041 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5101 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5161 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5221 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5281 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5341 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5401 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5461 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5521 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5581 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5641 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5701 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5761 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5821 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5881 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5941 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     6001 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6061 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6121 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6181 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6241 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6301 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6541 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     6841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     6901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     7021 cttttgctgg ccttttgctc acatgt
//
