LOCUS Exported 7542 bp ds-DNA circular SYN 30-AUG-2022 DEFINITION Gi-coupled hM4D DREADD fused with mCherry under the control of CaMKIIa promoter. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-hM4D(Gi)-mCherry SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7542) AUTHORS Bryan Roth TITLE Roth lab DREADDs JOURNAL Unpublished REFERENCE 2 (bases 1 to 7542) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 30, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7542 /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 1470..1479 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 2928..2937 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2934..3644 /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(3079..3097) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 3347..3366 /label=mCherry-F /note="mCherry, forward primer" misc_feature 3669..4257 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3722..3742) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4140..4151) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4289..4765 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4488..4507) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4805..4945 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4805..4934 /label=AAV2 ITR rep_origin 5020..5475 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5107..5126) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5317..5338 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5492..5511) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5611..5633 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5671..5689) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5757..5861 /gene="bla" /label=AmpR promoter CDS 5862..6722 /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(6080..6099) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6893..7481 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7382..7401 /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 ccaccatggc caacttcaca cctgtcaatg 1501 gcagctcggg caatcagtcc gtgcgcctgg tcacgtcatc atcccacaat cgctatgaga 1561 cggtggaaat ggtcttcatt gccacagtga caggctccct gagcctggtg actgtcgtgg 1621 gcaacatcct ggtgatgctg tccatcaagg tcaacaggca gctgcagaca gtcaacaact 1681 acttcctctt cagcctggcg tgtgctgatc tcatcatagg cgccttctcc atgaacctct 1741 acaccgtgta catcatcaag ggctactggc ccctgggcgc cgtggtctgc gacctgtggc 1801 tggccctgga ctgcgtggtg agcaacgcct ccgtcatgaa ccttctcatc atcagctttg 1861 accgctactt ctgcgtcacc aagcctctca cctaccctgc ccggcgcacc accaagatgg 1921 caggcctcat gattgctgct gcctgggtac tgtccttcgt gctctgggcg cctgccatct 1981 tgttctggca gtttgtggtg ggtaagcgga cggtgcccga caaccagtgc ttcatccagt 2041 tcctgtccaa cccagcagtg acctttggca cagccattgc tggcttctac ctgcctgtgg 2101 tcatcatgac ggtgctgtac atccacatct ccctggccag tcgcagccga gtccacaagc 2161 accggcccga gggcccgaag gagaagaaag ccaagacgct ggccttcctc aagagcccac 2221 taatgaagca gagcgtcaag aagcccccgc ccggggaggc cgcccgggag gagctgcgca 2281 atggcaagct ggaggaggcc cccccgccag cgctgccacc gccaccgcgc cccgtggctg 2341 ataaggacac ttccaatgag tccagctcag gcagtgccac ccagaacacc aaggaacgcc 2401 cagccacaga gctgtccacc acagaggcca ccacgcccgc catgcccgcc cctcccctgc 2461 agccgcgggc cctcaaccca gcctccagat ggtccaagat ccagattgtg acgaagcaga 2521 caggcaatga gtgtgtgaca gccattgaga ttgtgcctgc cacgccggct ggcatgcgcc 2581 ctgcggccaa cgtggcccgc aagttcgcca gcatcgctcg caaccaggtg cgcaagaagc 2641 ggcagatggc ggcccgggag cgcaaagtga cacgaacgat ctttgccatt ctgctggcct 2701 tcatcctcac ctggacgccc tacaacgtca tggtcctggt gaacaccttc tgccagagct 2761 gcatccctga cacggtgtgg tccattggct actggctctg ctacgtcaac agcaccatca 2821 accctgcctg ctatgctctg tgcaacgcca cctttaaaaa gaccttccgg cacctgctgc 2881 tgtgccagta tcggaacatc ggcactgcca ggcgggatcc accggtcgcc accatggtga 2941 gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc aaggtgcaca 3001 tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag ggccgcccct 3061 acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg cccttcgcct 3121 gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag caccccgccg 3181 acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag cgcgtgatga 3241 acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag gacggcgagt 3301 tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc gtaatgcaga 3361 agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac ggcgccctga 3421 agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac gctgaggtca 3481 agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac gtcaacatca 3541 agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac gaacgcgccg 3601 agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattc gatatcaagc 3661 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact 3721 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg 3781 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg 3841 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa 3901 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccggaact ttcgctttcc 3961 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg 4021 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt 4081 ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt 4141 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc 4201 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc 4261 gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc 4321 tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag 4381 ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt 4441 atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc 4501 aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc 4561 gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag 4621 ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa 4681 ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg 4741 aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg 4801 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact 4861 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc 4921 gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg 4981 tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat 5041 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag 5101 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc 5161 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc 5221 ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt 5281 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa 5341 caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg 5401 tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat 5461 taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 5521 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 5581 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 5641 cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta 5701 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg 5761 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat 5821 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc 5881 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa 5941 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac 6001 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga 6061 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag 6121 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca 6181 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca 6241 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa 6301 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc 6361 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa 6421 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag 6481 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct 6541 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac 6601 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa 6661 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt 6721 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat 6781 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg 6841 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc 6901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg 6961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag 7021 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact 7081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg 7141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc 7201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg 7261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg 7321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag 7381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc 7441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct 7501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gt //