LOCUS       Exported                7315 bp DNA     circular SYN 29-MAY-2024
DEFINITION  Double floxed Gq-coupled hM3D DREADD fused with mCherry under the 
            control of human synapsin promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn-DIO-hM3D(Gq)-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7315)
  AUTHORS   Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, 
            Maratos-Flier E, Roth BL, Lowell BB
  TITLE     Rapid, reversible activation of AgRP neurons drives feeding behavior
            in mice.
  JOURNAL   J Clin Invest. 2011 Apr;121(4):1424-8. doi: 10.1172/JCI46229.
  PUBMED    21364278
REFERENCE   2  (bases 1 to 7315)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7315)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 29, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName: 
            "J Clin Invest"; date: "2011-04"; volume: "121"; issue: "4"; pages: 
            "1424-8"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7315
                     /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    660..693
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    744..777
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(786..1496)
                     /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(1064..1083)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1333..1351
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1493..1502
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1518..3287)
                     /codon_start=1
                     /label=Muscarinic acetylcholine receptor M3
                     /translation="MTLHNNSTTSPLFPNISSSWIHSPSDAGLPPGTVTHFGSYNVSRA
                     AGNFSSPDGTTDDPLGGHTVWQVVFIAFLTGILALVTIIGNILVIVSFKVNKQLKTVNN
                     YFLLSLACADLIIGVISMNLFTTYIIMNRWALGNLACDLWLAIDCVASNASVMNLLVIS
                     FDRYFSITRPLTYRAKRTTKRAGVMIGLAWVISFVLWAPAILFWQYFVGKRTVPPGECF
                     IQFLSEPTITFGTAIAGFYMPVTIMTILYWRIYKETEKRTKELAGLQASGTEAETENFV
                     HPTGSSRSCSSYELQQQSMKRSNRRKYGRCHFWFTTKSWKPSSEQMDQDHSSSDSWNNN
                     DAAASLENSASSDEEDIGSETRAIYSIVLKLPGHSTILNSTKLPSSDNLQVPEEELGMV
                     DLERKADKLQAQKSVDDGGSFPKSFSKLPIQLESAVDTAKTSDVNSSVGKSTATLPLSF
                     KEATLAKRFALKTRSQITKRKRMSLVKEKKAAQTLSAILLAFIITWTPYNIMVLVNTFC
                     DSCIPKTFWNLGYWLCYINSTVNPVCYALCNKTFRTTFKMLLLCQCDKKKRRKQQYQQR
                     QSVIFHKRAPEQAL"
     protein_bind    complement(3300..3333)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(3384..3417)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    3442..4030
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(3495..3515)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3913..3924)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4062..4538
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4261..4280)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4578..4718
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4578..4707
                     /label=AAV2 ITR
     rep_origin      4793..5248
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4880..4899)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5090..5111
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5265..5284)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     5384..5406
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5444..5462)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        5530..5634
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5635..6495
                     /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(5853..5872)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6666..7254
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7155..7174
                     /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 gtaccggatc ctctagagtc gactccggaa
      661 taacttcgta taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc
      721 tattagcaat atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc
      781 gcgccttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
      841 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
      901 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
      961 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     1021 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     1081 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     1141 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     1201 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     1261 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     1321 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     1381 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     1441 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct caccatggtg
     1501 gcgaccgggg gatccttcaa ggcctgctcg ggtgcgcgct tgtgaaaaat gaccgactgt
     1561 ctctgctggt actgctgctt gcgcctcttt tttttgtcac actggcacag cagcagcatc
     1621 ttgaaagtgg ttctgaatgt tttgttgcac agagcatagc acacggggtt cacggtgctg
     1681 ttgatgtagc acagccagta gcccagattc caaaaggttt tgggtatgca gctgtcacaa
     1741 aaggtgttca ccagaaccat gatgttgtat ggggtccaag tgatgatgaa ggcaagcaag
     1801 atcgcactga gggtctgggc cgctttcttc tccttgacca gggacatcct tttccgctta
     1861 gtgatctgac ttctggtctt cagagcaaac ctcttggcca gagtggcttc cttgaaggac
     1921 agaggtagag tggccgtgct cttacccact gaggagttga cgtcagaagt cttagctgtg
     1981 tccacggctg actctagctg gatgggaagc ttggagaagc tttttggaaa actgcctcca
     2041 tcgtccacgc tcttctgggc ctgcagcttg tcggctttcc tctccaagtc caccatcccc
     2101 agctcctcct caggcacctg caggttgtcc gatgagggta acttggtgga gttgaggatg
     2161 gtgctgtgac ccggaagctt gagcacgatg gagtagatgg ctctcgtctc ggagccaatg
     2221 tcctcctcgt cggaggaggc ggagttctcc agggaggcag cagcatcatt gttgttccaa
     2281 ctgtcactgc tgctgtggtc ttggtccatc tgctcggagc tgggtttcca gctcttggtt
     2341 gtgaaccaga agtggcagcg gccatacttc ctcctgttgg agcgtttcat gctttgctgt
     2401 tgaagttcgt aactgctgca gcttcgagaa ctgcccgtgg ggtggacaaa gttttctgtc
     2461 tctgcctctg tcccagaggc ttgcaggcca gcaagctctt tggtacgctt ttcagtttcc
     2521 ttatagatcc tccagtataa aatagtcata atggtgacag gcatataaaa accagcgatg
     2581 gctgtgccaa aagtaatggt gggctcactg aggaactgaa tgaagcactc tcccggaggc
     2641 acagttctct ttccaacaaa gtattgccag aacaagatgg caggagccca aaggacaaag
     2701 gagatgaccc aagccagacc gatcatcaca ccggctctct ttgttgttcg tttggctcgg
     2761 tacgtgagcg gcctcgtgat ggaaaagtat ctgtcaaagc tgatgaccag aagattcata
     2821 acagaggcat tgctggctac gcagtcaatg gcaagccaga ggtcacaggc caagttccct
     2881 aaggcccatc gattcatgat gatgtaggtc gtaaacagat tcattgaaat gaccccgata
     2941 atcagatcgg cacaggccag gcttaagagg aagtagttgt tgaccgtctt cagctgcttg
     3001 ttgaccttaa atgacacaat taccaggatg ttgccgatga tggtcaccaa ggccaggatg
     3061 cccgttaaga aagcgatgaa gaccacttgc cagacggtat gacctcccag agggtcatcg
     3121 gtggtaccgt ctggagagga gaaattgcca gctgctcgag aaacattgta gctgccgaaa
     3181 tgagtgacgg ttcccggggg cagccctgca tcggaggggc tgtgtatcca ggaggagctg
     3241 atgtttggaa acaaaggcga ggttgtactg ttattgtgca aggtcatggt ggcgctagca
     3301 taacttcgta taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt
     3361 tattctttag aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa
     3421 ttcgatatca agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact
     3481 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg
     3541 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg
     3601 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg
     3661 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg
     3721 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc
     3781 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca
     3841 tcgtcctttc cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc
     3901 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct
     3961 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc
     4021 gcctccccgc atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc
     4081 cctccccagt gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc
     4141 taataaaatt aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt
     4201 ggaggggggt ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg
     4261 tctattggga accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct
     4321 cctgggttca agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca
     4381 tgaccaggct cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca
     4441 ggctggtctc caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg
     4501 gattacaggc gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt
     4561 gcggaccgag cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     4621 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     4681 ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct
     4741 ccttacgcat ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc
     4801 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     4861 gccagcgcct tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     4921 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     4981 cggcacctcg accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc
     5041 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     5101 ttccaaactg gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt
     5161 ttgccgattt cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     5221 tttaacaaaa tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat
     5281 gccgcatagt taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     5341 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     5401 cagaggtttt caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta
     5461 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5521 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5581 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5641 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5701 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5761 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5821 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5881 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5941 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6001 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6061 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6121 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     6181 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6241 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6301 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6361 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6421 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6481 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6541 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6601 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6661 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6721 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6781 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6841 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6901 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6961 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7021 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7081 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7141 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7201 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7261 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgt
//
