LOCUS Exported 6999 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gi-DREADD (P2A) nuclear dTomato expression in forebrain GABA-ergic interneurons under the control of the hDlx enhancer element. ACCESSION . VERSION . KEYWORDS pAAV-hDlx-GiDREADD-dTomato-Fishell-5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6999) AUTHORS Dimidschstein J, Chen Q, Tremblay R, Rogers SL, Saldi GA, Guo L, Xu Q, Liu R, Lu C, Chu J, Avery MC, Rashid MS, Baek M, Jacob AL, Smith GB, Wilson DE, Kosche G, Kruglikov I, Rusielewicz T, Kotak VC, Mowery TM, Anderson SA, Callaway EM, Dasen JS, Fitzpatrick D, Fossati V, Long MA, Noggle S, Reynolds JH, Sanes DH, Rudy B, Feng G, Fishell G TITLE A viral strategy for targeting and manipulating interneurons across vertebrate species. JOURNAL Nat Neurosci. 2016 Oct 31. doi: 10.1038/nn.4430. PUBMED 27798629 REFERENCE 2 (bases 1 to 6999) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6999 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR intron 860..992 /label=chimeric intron /note="chimera between introns from human beta-globin and immunoglobulin heavy chain genes" primer_bind 975..994 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 1035..1044 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2502..2558 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 2571..2591 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 2592..3290 /codon_start=1 /product="dimeric variant of DsRed fluorescent protein (Shaner et al., 2004)" /label=dTomato /note="mammalian codon-optimized" /translation="VSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGL VTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIHQ ALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRHH LFLYGMDELYK" primer_bind complement(2722..2740) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2990..3009 /label=mCherry-F /note="mCherry, forward primer" primer_bind 3184..3207 /label=DsRed1-C /note="DsRed1, forward primer" misc_feature 3345..3933 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3398..3418) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3816..3827) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 4102..4183 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4139..4158) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4193..4212 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" repeat_region 4262..4402 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4262..4391 /label=AAV2 ITR rep_origin 4477..4932 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4564..4583) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4774..4795 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4949..4968) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5068..5090 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5128..5146) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5214..5318 /gene="bla" /label=AmpR promoter CDS 5319..6179 /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(5537..5556) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6350..6938 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6839..6858 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtttatt cagaatgtta tgcactcaca 181 gtggtttggc atgcatctgg tgaatttttt ttaacgaaaa attagtgttg gtttcgatgt 241 atggtagcat tctccctaac gtaatttgaa taattcagca aagccccact accagctgta 301 cttctgcagc ctcttccatt cttttcagca ttataatttt ggttaatttt caattttagg 361 tcctacgtct ctgcaatttg tgtatgaata acagaataat ttccctcttt tgtttcgcct 421 ttcctgttcc tgaatctaaa taaagatggc tttttagtat taaaagtgga agaaaattac 481 aggtaattat ctttgacggt aaaaacgctg taatcagcgg gctacatgaa aaattactct 541 aattatggct gcatttaaga gaatggaaaa aaaccttctt gtggataaaa accttaaatt 601 gtccccaatg tctgcttcaa attggatggc actgcagctg gaggctttgt tcagaattga 661 tcctggggag ctacgaaccc aaagtttcac agtaggaagc ttgatatcga attcctgcag 721 cccgggctgg gcataaaagt cagggcagag ccatctattg cttacatttg cttctagcct 781 gcaggtcgag gagcgcagcc ttccagaagc agagcgcggc gccttaagct gcagaagttg 841 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata 901 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct 961 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc 1021 ttaagaaagc tagcgccacc atggccaact tcacacctgt caatggcagc tcgggcaatc 1081 agtccgtgcg cctggtcacg tcatcatccc acaatcgcta tgagacggtg gaaatggtct 1141 tcattgccac agtgacaggc tccctgagcc tggtgactgt cgtgggcaac atcctggtga 1201 tgctgtccat caaggtcaac aggcagctgc agacagtcaa caactacttc ctcttcagcc 1261 tggcgtgtgc tgatctcatc ataggcgcct tctccatgaa cctctacacc gtgtacatca 1321 tcaagggcta ctggcccctg ggcgccgtgg tctgcgacct gtggctggcc ctggactgcg 1381 tggtgagcaa cgcctccgtc atgaaccttc tcatcatcag ctttgaccgc tacttctgcg 1441 tcaccaagcc tctcacctac cctgcccggc gcaccaccaa gatggcaggc ctcatgattg 1501 ctgctgcctg ggtactgtcc ttcgtgctct gggcgcctgc catcttgttc tggcagtttg 1561 tggtgggtaa gcggacggtg cccgacaacc agtgcttcat ccagttcctg tccaacccag 1621 cagtgacctt tggcacagcc attgctggct tctacctgcc tgtggtcatc atgacggtgc 1681 tgtacatcca catctccctg gccagtcgca gccgagtcca caagcaccgg cccgagggcc 1741 cgaaggagaa gaaagccaag acgctggcct tcctcaagag cccactaatg aagcagagcg 1801 tcaagaagcc cccgcccggg gaggccgccc gggaggagct gcgcaatggc aagctggagg 1861 aggccccccc gccagcgctg ccaccgccac cgcgccccgt ggctgataag gacacttcca 1921 atgagtccag ctcaggcagt gccacccaga acaccaagga acgcccagcc acagagctgt 1981 ccaccacaga ggccaccacg cccgccatgc ccgcccctcc cctgcagccg cgggccctca 2041 acccagcctc cagatggtcc aagatccaga ttgtgacgaa gcagacaggc aatgagtgtg 2101 tgacagccat tgagattgtg cctgccacgc cggctggcat gcgccctgcg gccaacgtgg 2161 cccgcaagtt cgccagcatc gctcgcaacc aggtgcgcaa gaagcggcag atggcggccc 2221 gggagcgcaa agtgacacga acgatctttg ccattctgct ggccttcatc ctcacctgga 2281 cgccctacaa cgtcatggtc ctggtgaaca ccttctgcca gagctgcatc cctgacacgg 2341 tgtggtccat tggctactgg ctctgctacg tcaacagcac catcaaccct gcctgctatg 2401 ctctgtgcaa cgccaccttt aaaaagacct tccggcacct gctgctgtgc cagtatcgga 2461 acatcggcac tgccaggcgg tccggagata acggaagcgg agccactaac ttctccctgt 2521 tgaaacaagc aggggatgtc gaagagaatc ccgggccatc tagaatggcc cccaagaaga 2581 agaggaaggt ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc ttcaaggtgc 2641 gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc 2701 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc ctgcccttcg 2761 cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg aagcaccccg 2821 ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga 2881 tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg caggacggca 2941 cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc cccgtaatgc 3001 agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc gacggcgtgc 3061 tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac ctggtggagt 3121 tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac tacgtggaca 3181 ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag tacgagcgct 3241 ccgagggccg ccaccacctg ttcctgtacg gcatggacga gctgtacaag taagtcgacg 3301 gcgcgcccca cccctgcagg gaattcgata tcaagcttat cgataatcaa cctctggatt 3361 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg 3421 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct 3481 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc 3541 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca 3601 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac 3661 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt 3721 ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctat gttgccacct 3781 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc 3841 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga 3901 cgagtcggat ctccctttgg gccgcctccc cgcatcgata ccgagcgctg ctcgagagat 3961 ctgtgatagc ggccatcaag ctggccgcga ctctagatca taatcagcca taccacattt 4021 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 4081 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 4141 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 4201 tccaaactca tcaatgtatc agcttatcga taccgcatgc acgtgcggac cgagcggccg 4261 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag 4321 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag 4381 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc 4441 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa 4501 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc 4561 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag 4621 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca 4681 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc 4741 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa 4801 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct 4861 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa 4921 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc 4981 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat 5041 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt 5101 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg 5161 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa 5221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac 5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg 5341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc 5401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg 5461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga 5521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc 5581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag 5641 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga 5701 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg 5761 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga 5821 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt 5881 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact 5941 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt 6001 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg 6061 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta 6121 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac 6181 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta 6241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt 6301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt 6361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt 6421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc 6481 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg 6541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg 6601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt 6661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac 6721 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg 6781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg 6841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat 6901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt 6961 tacggttcct ggccttttgc tggccttttg ctcacatgt //