LOCUS Exported 7875 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gq-coupled hM3D DREADD fused with mCherry under the control of CaMKIIa promoter. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-hM3D(Gq)-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7875) AUTHORS Bryan Roth TITLE Roth lab DREADDs JOURNAL Unpublished REFERENCE 2 (bases 1 to 7875) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7875 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 149..1437 /label=CaMKII promoter /note="promoter for the mouse calcium/calmodulin-dependent kinase II alpha subunit" regulatory 3261..3270 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3267..3977 /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(3412..3430) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3680..3699 /label=mCherry-F /note="mCherry, forward primer" misc_feature 4002..4590 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4055..4075) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4473..4484) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4622..5098 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4821..4840) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 5138..5278 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5138..5267 /label=AAV2 ITR rep_origin 5353..5808 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5440..5459) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5650..5671 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5825..5844) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5944..5966 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6004..6022) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6090..6194 /gene="bla" /label=AmpR promoter CDS 6195..7055 /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(6413..6432) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7226..7814 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7715..7734 /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 gcgtttaaca ttatggcctt aggtcacttc atctccatgg 181 ggttcttctt ctgattttct agaaaatgag atgggggtgc agagagcttc ctcagtgacc 241 tgcccagggt cacatcagaa atgtcagagc tagaacttga actcagatta ctaatcttaa 301 attccatgcc ttgggggcat gcaagtacga tatacagaag gagtgaactc attagggcag 361 atgaccaatg agtttaggaa agaagagtcc agggcagggt acatctacac cacccgccca 421 gccctgggtg agtccagcca cgttcacctc attatagttg cctctctcca gtcctacctt 481 gacgggaagc acaagcagaa actgggacag gagccccagg agaccaaatc ttcatggtcc 541 ctctgggagg atgggtgggg agagctgtgg cagaggcctc aggaggggcc ctgctgctca 601 gtggtgacag ataggggtga gaaagcagac agagtcattc cgtcagcatt ctgggtctgt 661 ttggtacttc ttctcacgct aaggtggcgg tgtgatatgc acaatggcta aaaagcaggg 721 agagctggaa agaaacaagg acagagacag aggccaagtc aaccagacca attcccagag 781 gaagcaaaga aaccattaca gagactacaa gggggaaggg aaggagagat gaattagctt 841 cccctgtaaa ccttagaacc cagctgttgc cagggcaacg gggcaatacc tgtctcttca 901 gaggagatga agttgccagg gtaactacat cctgtctttc tcaaggacca tcccagaatg 961 tggcacccac tagccgttac catagcaact gcctctttgc cccacttaat cccatcccgt 1021 ctgttaaaag ggccctatag ttggaggtgg gggaggtagg aagagcgatg atcacttgtg 1081 gactaagttt gttcgcatcc ccttctccaa ccccctcagt acatcaccct gggggaacag 1141 ggtccacttg ctcctgggcc cacacagtcc tgcagtattg tgtatataag gccagggcaa 1201 agaggagcag gttttaaagt gaaaggcagg caggtgttgg ggaggcagtt accggggcaa 1261 cgggaacagg gcgtttcgga ggtggttgcc atggggacct ggatgctgac gaaggctcgc 1321 gaggctgtga gcagccacag tgccctgctc agaagcccca agctcgtcag tcaagccggt 1381 tctccgtttg cactcaggag cacgggcagg cgagtggccc ctagttctgg gggcagctct 1441 agagcggtac cggatcctct agagtcgacg ccaccatgac cttgcacaat aacagtacaa 1501 cctcgccttt gtttccaaac atcagctcct cctggataca cagcccctcc gatgcagggc 1561 tgcccccggg aaccgtcact catttcggca gctacaatgt ttctcgagca gctggcaatt 1621 tctcctctcc agacggtacc accgatgacc ctctgggagg tcataccgtc tggcaagtgg 1681 tcttcatcgc tttcttaacg ggcatcctgg ccttggtgac catcatcggc aacatcctgg 1741 taattgtgtc atttaaggtc aacaagcagc tgaagacggt caacaactac ttcctcttaa 1801 gcctggcctg tgccgatctg attatcgggg tcatttcaat gaatctgttt acgacctaca 1861 tcatcatgaa tcgatgggcc ttagggaact tggcctgtga cctctggctt gccattgact 1921 gcgtagccag caatgcctct gttatgaatc ttctggtcat cagctttgac agatactttt 1981 ccatcacgag gccgctcacg taccgagcca aacgaacaac aaagagagcc ggtgtgatga 2041 tcggtctggc ttgggtcatc tcctttgtcc tttgggctcc tgccatcttg ttctggcaat 2101 actttgttgg aaagagaact gtgcctccgg gagagtgctt cattcagttc ctcagtgagc 2161 ccaccattac ttttggcaca gccatcgctg gtttttatat gcctgtcacc attatgacta 2221 ttttatactg gaggatctat aaggaaactg aaaagcgtac caaagagctt gctggcctgc 2281 aagcctctgg gacagaggca gagacagaaa actttgtcca ccccacgggc agttctcgaa 2341 gctgcagcag ttacgaactt caacagcaaa gcatgaaacg ctccaacagg aggaagtatg 2401 gccgctgcca cttctggttc acaaccaaga gctggaaacc cagctccgag cagatggacc 2461 aagaccacag cagcagtgac agttggaaca acaatgatgc tgctgcctcc ctggagaact 2521 ccgcctcctc cgacgaggag gacattggct ccgagacgag agccatctac tccatcgtgc 2581 tcaagcttcc gggtcacagc accatcctca actccaccaa gttaccctca tcggacaacc 2641 tgcaggtgcc tgaggaggag ctggggatgg tggacttgga gaggaaagcc gacaagctgc 2701 aggcccagaa gagcgtggac gatggaggca gttttccaaa aagcttctcc aagcttccca 2761 tccagctaga gtcagccgtg gacacagcta agacttctga cgtcaactcc tcagtgggta 2821 agagcacggc cactctacct ctgtccttca aggaagccac tctggccaag aggtttgctc 2881 tgaagaccag aagtcagatc actaagcgga aaaggatgtc cctggtcaag gagaagaaag 2941 cggcccagac cctcagtgcg atcttgcttg ccttcatcat cacttggacc ccatacaaca 3001 tcatggttct ggtgaacacc ttttgtgaca gctgcatacc caaaaccttt tggaatctgg 3061 gctactggct gtgctacatc aacagcaccg tgaaccccgt gtgctatgct ctgtgcaaca 3121 aaacattcag aaccactttc aagatgctgc tgctgtgcca gtgtgacaaa aaaaagaggc 3181 gcaagcagca gtaccagcag agacagtcgg tcatttttca caagcgcgca cccgagcagg 3241 ccttgaagga tcccccggtc gccaccatgg tgagcaaggg cgaggaggat aacatggcca 3301 tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg ctccgtgaac ggccacgagt 3361 tcgagatcga gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga 3421 aggtgaccaa gggtggcccc ctgcccttcg cctgggacat cctgtcccct cagttcatgt 3481 acggctccaa ggcctacgtg aagcaccccg ccgacatccc cgactacttg aagctgtcct 3541 tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga ggacggcggc gtggtgaccg 3601 tgacccagga ctcctccctg caggacggcg agttcatcta caaggtgaag ctgcgcggca 3661 ccaacttccc ctccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcct 3721 ccgagcggat gtaccccgag gacggcgccc tgaagggcga gatcaagcag aggctgaagc 3781 tgaaggacgg cggccactac gacgctgagg tcaagaccac ctacaaggcc aagaagcccg 3841 tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga catcacctcc cacaacgagg 3901 actacaccat cgtggaacag tacgaacgcg ccgagggccg ccactccacc ggcggcatgg 3961 acgagctgta caagtaagaa ttcgatatca agcttatcga taatcaacct ctggattaca 4021 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 4081 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 4141 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 4201 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 4261 cctgtcagct cctttccgga actttcgctt tccccctccc tattgccacg gcggaactca 4321 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 4381 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga 4441 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 4501 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 4561 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta 4621 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc 4681 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc 4741 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac 4801 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg 4861 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt 4921 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg 4981 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc 5041 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct 5101 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg 5161 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga 5221 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg 5281 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa 5341 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc 5401 gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct ttcttccctt 5461 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag 5521 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt 5581 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt 5641 tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc tcgggctatt 5701 cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat gagctgattt 5761 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc 5821 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg 5881 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg 5941 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa 6001 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga 6061 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 6121 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 6181 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 6241 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 6301 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 6361 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 6421 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 6481 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 6541 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 6601 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 6661 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 6721 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 6781 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 6841 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 6901 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 6961 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 7021 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 7081 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 7141 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 7201 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 7261 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 7321 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 7381 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 7441 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 7501 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 7561 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 7621 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 7681 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 7741 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 7801 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 7861 cttttgctca catgt //