LOCUS Exported 7935 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Double floxed Gq-coupled hM3D-IRES-mCitrine under the control of human synapsin promoter. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-DIO-HA-hM3D(Gq)-IRES-mCitrine SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7935) AUTHORS Bryan Roth TITLE Roth lab DREADDs JOURNAL Unpublished REFERENCE 2 (bases 1 to 7935) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7935 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 7..136 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 181..628 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind 666..699 /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 750..783 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(798..1514) /codon_start=1 /product="enhanced variant of YFP (Heikal et al., 2001)" /label=Citrine /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(840..861) /label=EGFP-C /note="EGFP, forward primer" primer_bind 1189..1208 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1448..1469 /label=EGFP-N /note="EGFP, reverse primer" regulatory 1511..1520 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" misc_feature 1521..2094 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1681..1700) /label=IRES-F /note="IRES internal ribosome entry site, forward primer" primer_bind 1910..1927 /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" CDS complement(3880..3906) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" protein_bind complement(3920..3953) /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(4004..4037) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 4062..4650 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4115..4135) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4533..4544) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4682..5158 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4881..4900) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 5198..5338 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5198..5327 /label=AAV2 ITR rep_origin 5413..5868 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5500..5519) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5710..5731 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5885..5904) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6004..6026 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6064..6082) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6150..6254 /gene="bla" /label=AmpR promoter CDS 6255..7115 /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(6473..6492) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7286..7874 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7775..7794 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cnsnsscctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg 61 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc 121 atcactaggg gttcctgcgg ccgcacgcgt gtgtctagac tgcagagggc cctgcgtatg 181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga 241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga 301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg 361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc 421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg 481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg 541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg 601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggtac cggatcctct agagtcgact 661 ccggaataac ttcgtatagg atactttata cgaagttatg cagaatggta gctggattgt 721 agctgctatt agcaatatga aacctcttaa taacttcgta tagcatacat tatacgaagt 781 tatggcgcgc cctatcactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc 841 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagtttggac 901 tggtagctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc 961 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag 1021 ttcaccttga tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag 1081 ttgtactcca gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc 1141 tcgatgcggt tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg 1201 ccgtcgtcct tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg 1261 aagaagtcgt gctgcttcat gtggtcgggg tagcgggcga agcacatcag gccgtagccg 1321 aaggtggtca cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc 1381 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg 1441 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg 1501 cccttgctca ccatggtggc ttatcatcgt gtttttcaaa ggaaaaccac gtccccgtgg 1561 ttcggggggc ctagacgttt ttttaacctc gactaaacac atgtaaagca tgtgcaccga 1621 ggccccagat cagatcccat acaatggggt accttctggg catccttcag ccccttgttg 1681 aatacgcttg aggagagcca tttgactctt tccacaacta tccaactcac aacgtggcac 1741 tggggttgtg ccgcctttgc aggtgtatct tatacacgtg gcttttggcc gcagaggcac 1801 ctgtcgccag gtggggggtt ccgctgcctg caaagggtcg ctacagacgt tgtttgtctt 1861 caagaagctt ccagaggaac tgcttccttc acgacattca acagaccttg cattcctttg 1921 gcgagagggg aaagacccct aggaatgctc gtcaagaaga cagggccagg tttccgggcc 1981 ctcacattgc caaaagacgg caatatggtg gaaaataaca tatagacaaa cgcacaccgg 2041 ccttattcca agcggcttcg gccagtaacg ttaggggggg gggagggaga ggggcgatat 2101 cgcggccgcc tacaaggcct gctcgggtgc gcgcttgtga aaaatgaccg actgtctctg 2161 ctggtactgc tgcttgcgcc tctttttttt gtcacactgg cacagcagca gcatcttgaa 2221 agtggttctg aatgttttgt tgcacagagc atagcacacg gggttcacgg tgctgttgat 2281 gtagcacagc cagtagccca gattccaaaa ggttttgggt atgcagctgt cacaaaaggt 2341 gttcaccaga accatgatgt tgtatggggt ccaagtgatg atgaaggcaa gcaagatcgc 2401 actgagggtc tgggccgctt tcttctcctt gaccagggac atccttttcc gcttagtgat 2461 ctgacttctg gtcttcagag caaacctctt ggccagagtg gcttccttga aggacagagg 2521 tagagtggcc gtgctcttac ccactgagga gttgacgtca gaagtcttag ctgtgtccac 2581 ggctgactct agctggatgg gaagcttgga gaagcttttt ggaaaactgc ctccatcgtc 2641 cacgctcttc tgggcctgca gcttgtcggc tttcctctcc aagtccacca tccccagctc 2701 ctcctcaggc acctgcaggt tgtccgatga gggtaacttg gtggagttga ggatggtgct 2761 gtgacccgga agcttgagca cgatggagta gatggctctc gtctcggagc caatgtcctc 2821 ctcgtcggag gaggcggagt tctccaggga ggcagcagca tcattgttgt tccaactgtc 2881 actgctgctg tggtcttggt ccatctgctc ggagctgggt ttccagctct tggttgtgaa 2941 ccagaagtgg cagcggccat acttcctcct gttggagcgt ttcatgcttt gctgttgaag 3001 ttcgtaactg ctgcagcttc gagaactgcc cgtggggtgg acaaagtttt ctgtctctgc 3061 ctctgtccca gaggcttgca ggccagcaag ctctttggta cgcttttcag tttccttata 3121 gatcctccag tataaaatag tcataatggt gacaggcata taaaaaccag cgatggctgt 3181 gccaaaagta atggtgggct cactgaggaa ctgaatgaag cactctcccg gaggcacagt 3241 tctctttcca acaaagtatt gccagaacaa gatggcagga gcccaaagga caaaggagat 3301 gacccaagcc agaccgatca tcacaccggc tctctttgtt gttcgtttgg ctcggtacgt 3361 gagcggcctc gtgatggaaa agtatctgtc aaagctgatg accagaagat tcataacaga 3421 ggcattgctg gctacgcagt caatggcaag ccagaggtca caggccaagt tccctaaggc 3481 ccatcgattc atgatgatgt aggtcgtaaa cagattcatt gaaatgaccc cgataatcag 3541 atcggcacag gccaggctta agaggaagta gttgttgacc gtcttcagct gcttgttgac 3601 cttaaatgac acaattacca ggatgttgcc gatgatggtc accaaggcca ggatgcccgt 3661 taagaaagcg atgaagacca cttgccagac ggtatgacct cccagagggt catcggtggt 3721 accgtctgga gaggagaaat tgccagctgc tcgagaaaca ttgtagctgc cgaaatgagt 3781 gacggttccc gggggcagcc ctgcatcgga ggggctgtgt atccaggagg agctgatgtt 3841 tggaaacaaa ggcgaggttg tactgttatt gtgcaaggta gcgtaatctg gaacatcgta 3901 tgggtacatg gtggctagca taacttcgta taaagtatcc tatacgaagt tatttgcctt 3961 aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt cgtataatgt 4021 atgctatacg aagttatgaa ttcgatatca agcttatcga taatcaacct ctggattaca 4081 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 4141 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 4201 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 4261 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 4321 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 4381 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 4441 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga 4501 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 4561 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 4621 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta 4681 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc 4741 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc 4801 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac 4861 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg 4921 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt 4981 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg 5041 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc 5101 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct 5161 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg 5221 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga 5281 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg 5341 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa 5401 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 5461 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt 5521 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 5581 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt 5641 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 5701 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt 5761 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt 5821 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc 5881 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg 5941 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg 6001 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa 6061 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga 6121 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 6181 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 6241 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 6301 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 6361 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 6421 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 6481 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 6541 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 6601 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 6661 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 6721 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 6781 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 6841 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 6901 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 6961 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 7021 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 7081 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 7141 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 7201 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 7261 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 7321 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 7381 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 7441 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 7501 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 7561 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 7621 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 7681 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 7741 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 7801 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 7861 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 7921 cttttgctca catgt //