LOCUS Exported 8476 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gq-coupled hM3D-IRES-mCitrine under the control of CaMKIIa promoter. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-HA-hM3D(Gq)-IRES-mCitrine SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8476) AUTHORS Bryan Roth TITLE Roth lab DREADDs JOURNAL Unpublished REFERENCE 2 (bases 1 to 8476) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8476 /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" CDS 1467..1493 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 3279..3852 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(3446..3463) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 3673..3692 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" regulatory 3853..3862 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3859..4575 /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(3904..3925) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4165..4184) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4512..4533 /label=EGFP-C /note="EGFP, forward primer" misc_feature 4603..5191 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4656..4676) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5074..5085) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 5223..5699 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(5422..5441) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 5739..5879 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5739..5868 /label=AAV2 ITR rep_origin 5954..6409 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6041..6060) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6251..6272 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(6426..6445) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 6545..6567 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6605..6623) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 6691..6795 /gene="bla" /label=AmpR promoter CDS 6796..7656 /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(7014..7033) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7827..8415 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8316..8335 /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 cggatccgcc accatgtacc catacgatgt tccagattac gctaccttgc 1501 acaataacag tacaacctcg cctttgtttc caaacatcag ctcctcctgg atacacagcc 1561 cctccgatgc agggctgccc ccgggaaccg tcactcattt cggcagctac aatgtttctc 1621 gagcagctgg caatttctcc tctccagacg gtaccaccga tgaccctctg ggaggtcata 1681 ccgtctggca agtggtcttc atcgctttct taacgggcat cctggccttg gtgaccatca 1741 tcggcaacat cctggtaatt gtgtcattta aggtcaacaa gcagctgaag acggtcaaca 1801 actacttcct cttaagcctg gcctgtgccg atctgattat cggggtcatt tcaatgaatc 1861 tgtttacgac ctacatcatc atgaatcgat gggccttagg gaacttggcc tgtgacctct 1921 ggcttgccat tgactgcgta gccagcaatg cctctgttat gaatcttctg gtcatcagct 1981 ttgacagata cttttccatc acgaggccgc tcacgtaccg agccaaacga acaacaaaga 2041 gagccggtgt gatgatcggt ctggcttggg tcatctcctt tgtcctttgg gctcctgcca 2101 tcttgttctg gcaatacttt gttggaaaga gaactgtgcc tccgggagag tgcttcattc 2161 agttcctcag tgagcccacc attacttttg gcacagccat cgctggtttt tatatgcctg 2221 tcaccattat gactatttta tactggagga tctataagga aactgaaaag cgtaccaaag 2281 agcttgctgg cctgcaagcc tctgggacag aggcagagac agaaaacttt gtccacccca 2341 cgggcagttc tcgaagctgc agcagttacg aacttcaaca gcaaagcatg aaacgctcca 2401 acaggaggaa gtatggccgc tgccacttct ggttcacaac caagagctgg aaacccagct 2461 ccgagcagat ggaccaagac cacagcagca gtgacagttg gaacaacaat gatgctgctg 2521 cctccctgga gaactccgcc tcctccgacg aggaggacat tggctccgag acgagagcca 2581 tctactccat cgtgctcaag cttccgggtc acagcaccat cctcaactcc accaagttac 2641 cctcatcgga caacctgcag gtgcctgagg aggagctggg gatggtggac ttggagagga 2701 aagccgacaa gctgcaggcc cagaagagcg tggacgatgg aggcagtttt ccaaaaagct 2761 tctccaagct tcccatccag ctagagtcag ccgtggacac agctaagact tctgacgtca 2821 actcctcagt gggtaagagc acggccactc tacctctgtc cttcaaggaa gccactctgg 2881 ccaagaggtt tgctctgaag accagaagtc agatcactaa gcggaaaagg atgtccctgg 2941 tcaaggagaa gaaagcggcc cagaccctca gtgcgatctt gcttgccttc atcatcactt 3001 ggaccccata caacatcatg gttctggtga acaccttttg tgacagctgc atacccaaaa 3061 ccttttggaa tctgggctac tggctgtgct acatcaacag caccgtgaac cccgtgtgct 3121 atgctctgtg caacaaaaca ttcagaacca ctttcaagat gctgctgctg tgccagtgtg 3181 acaaaaaaaa gaggcgcaag cagcagtacc agcagagaca gtcggtcatt tttcacaagc 3241 gcgcacccga gcaggccttg taggcggccg cgatatcgcc cctctccctc ccccccccct 3301 aacgttactg gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt 3361 tccaccatat tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg 3421 acgagcattc ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc 3481 gtgaaggaag cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt 3541 tgcaggcagc ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta 3601 taagatacac ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg 3661 gaaagagtca aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag 3721 gtaccccatt gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag 3781 tcgaggttaa aaaaacgtct aggccccccg aaccacgggg acgtggtttt cctttgaaaa 3841 acacgatgat aagccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc 3901 atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc 3961 gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg 4021 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc 4081 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc 4141 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag 4201 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac 4261 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg 4321 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac 4381 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg 4441 ctgctgcccg acaaccacta cctgagctac cagtccaaac tgagcaaaga ccccaacgag 4501 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg 4561 gacgagctgt acaagtgaga attcgatatc aagcttatcg ataatcaacc tctggattac 4621 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 4681 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 4741 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 4801 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 4861 acctgtcagc tcctttccgg aactttcgct ttccccctcc ctattgccac ggcggaactc 4921 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 4981 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg 5041 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 5101 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 5161 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct 5221 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc 5281 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt 5341 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga 5401 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt 5461 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt 5521 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac 5581 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac 5641 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc 5701 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt 5761 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg 5821 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg 5881 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca 5941 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 6001 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct 6061 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 6121 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt 6181 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 6241 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat 6301 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt 6361 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact 6421 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 6481 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 6541 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 6601 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 6661 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 6721 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 6781 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 6841 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 6901 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 6961 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 7021 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 7081 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 7141 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 7201 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 7261 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 7321 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 7381 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 7441 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 7501 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 7561 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 7621 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 7681 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 7741 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 7801 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 7861 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 7921 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 7981 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 8041 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 8101 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 8161 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 8221 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 8281 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 8341 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 8401 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 8461 ccttttgctc acatgt //