LOCUS pAAV-hsyn-GRAB_5 6530 bp DNA circular SYN 09-APR-2021 DEFINITION Expresses the 5-HT sensor GRAB_5-HT1.0 in neurons. ACCESSION . VERSION . KEYWORDS pAAV-hsyn-GRAB_5-HT1.0. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6530) TITLE Yulong Li 5-HT sensor and DA sensor plasmids REFERENCE 2 (bases 1 to 6530) TITLE Direct Submission REFERENCE 3 (bases 1 to 6530) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..6530 /mol_type="other DNA" /organism="synthetic DNA construct" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="AAV2 ITR (alternate)" /note="Functional equivalent of wild-type AAV2 ITR" promoter 145..586 /note="synapsin" /note="human synapsin promoter is a neuron-specific promoter" regulatory 581..590 /label=Kozak sequence /note="Kozak sequence" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" CDS 587..649 /codon_start=1 /product="leader sequence from mouse immunoglobulin kappa light chain" /label=leader sequence from mouse immunoglobulin kappa /note="Ig-kappa leader" /translation="METDTLLLWVLLLWVPGSTGD" protein_bind 650..674 /gene="mutant version of attB" /label=BP Clonase(TM) binding site /bound_moiety="BP Clonase(TM)" /note="attB1" /note="recombination site for the Gateway(R) BP reaction" CDS 683..1420 /codon_start=1 /label=Human HTR2C(2-247) /note="Human HTR2C(2-247)" /note="G-protein coupled receptor for 5-hydroxytryptamine (serotonin). Also functions as a receptor for various drugs and psychoactive substances, including ergot alkaloid derivatives, 1-2,5,-dimethoxy-4-iodophenyl-2-aminopropane (DOI) and lysergic acid diethylamide (LSD). Ligand binding causes a conformation change that triggers signaling via guanine nucleotide-binding proteins (G proteins) and modulates the activity of down-stream effectors. Beta-arrestin family members inhibit signaling via G proteins and mediate activation of alternative signaling pathways. Signaling activates a phosphatidylinositol-calcium second messenger system that modulates the activity of phosphatidylinositol 3-kinase and down-stream signaling cascades and promotes the release of Ca2+ ions from intracellular stores. Regulates neuronal activity via the activation of short transient receptor potential calcium channels in the brain, and thereby modulates the activation of pro-opiomelacortin neurons and the release of CRH that then regulates the release of corticosterone. Plays a role in the regulation of appetite and eating behavior, responses to anxiogenic stimuli and stress. Plays a role in insulin sensitivity and glucose homeostasis." /translation="VNLRNAVHSFLVHLIGLLVWQCDISVSPVAAIVTDIFNTSDGGRF KFPDGVQNWPALSIVIIIIMTIGGNILVIMAVSMEKKLHNATNYFLMSLAIADMLVGLL VMPLSLLAILYDYVWPLPRYLCPVWISLDVLFSTASIMHLCAISLDRYVAIRNPIEHSR FNSRTKAIMKIAIVWAISIGVSVPIPVIGLRDEEKVFVNNTTCVLNDPNFVLIGSFVAF FIPLTIMVITYCLTIYVLRRQALM" CDS 1451..1702 /label=VC155 /note="C-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" CDS 2177..2644 /codon_start=1 /label=Human HTR2C(304-458) /note="Human HTR2C(304-458)" /translation="NNERKASKVLGIVFFVFLIMWCPFFITNILSVLCEKSCNQKLMEK LLNVFVWIGYVCSGINPLVYTLFNKIYRRAFSNYLRCNYKVEKKPPVRQIPRVAATALS GRELNVNIYRHTNEPVIEKASDNEPGIEMQVENLELPVNPSSVVSERISSV" misc_feature 2657..3245 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" polyA_signal 3277..3753 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" repeat_region 3793..3933 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" rep_origin 4008..4463 /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 4745..4849 /label=AmpR promoter CDS 4850..5707 /label=AmpR /note="beta-lactamase" rep_origin 5881..6469 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtatgagg cggggtgggg gtgcctacct gacgaccgac 181 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag 241 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg 301 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg 361 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc 421 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc 481 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga 541 ggagtcgtgt cgtgcctgag agcgcagtcg agaaaccggt gccaccatgg agacagacac 601 actcctgcta tgggtactgc tgctctgggt tccaggttcc actggtgaca caagtttgta 661 caaaaaagtt ggcaccaccg gtgtgaacct gaggaatgcg gtgcattcat tccttgtgca 721 cctaattggc ctattggttt ggcaatgtga tatttctgtg agcccagtag cagctatagt 781 aactgacatt ttcaatacct ccgatggtgg acgcttcaaa ttcccagacg gggtacaaaa 841 ctggccagca ctttcaatcg tcatcataat aatcatgaca ataggtggca acatccttgt 901 gatcatggca gtaagcatgg aaaagaaact gcacaatgcc accaattact tcttaatgtc 961 cctagccatt gctgatatgc tagtgggact acttgtcatg cccctgtctc tcctggcaat 1021 cctttatgat tatgtctggc cactacctag atatttgtgc cccgtctgga tttctttaga 1081 tgttttattt tcaacagcgt ccatcatgca cctctgcgct atatcgctgg atcggtatgt 1141 agcaatacgt aatcctattg agcatagccg tttcaattcg cggactaagg ccatcatgaa 1201 gattgctatt gtttgggcaa tttctatagg tgtatcagtt cctatccctg tgattggact 1261 gagggacgaa gaaaaggtgt tcgtgaacaa cacgacgtgc gtgctcaacg acccaaattt 1321 cgttcttatt gggtccttcg tagctttctt cataccgctg acgattatgg tgattacgta 1381 ttgcctgacc atctacgttc tgcgccgaca agctttgatg tttctgaacg gaaacgtcta 1441 tatcaaggcc gacaagcaga agaacggcat caaggcgaac ttccacatcc gccacaacat 1501 cgaggacggc ggcgtgcagc tcgcctacca ctaccagcag aacaccccca tcggcgacgg 1561 ccccgtgctg ctgcccgaca accactacct gagcgtgcag tccaaacttt cgaaagaccc 1621 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg gaatcactct 1681 cggcatggac gagctgtaca agggcggtac cggagggcaa gaatccatga gcagcaaggg 1741 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 1801 ccacaagttc agcgtgtccg gcgagggtga gggcgatgcc accactggca agctgaccct 1861 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 1921 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt 1981 caagtccgcc atgcccgaag gctacatcca ggagcgcacc atcttcttca aggacgacgg 2041 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 2101 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 2161 cggatttgca acagcaaaca atgaaagaaa agcttcgaaa gtccttggga ttgttttctt 2221 tgtgtttctg atcatgtggt gcccattttt cattaccaat attctgtctg ttctttgtga 2281 gaagtcctgt aaccaaaagc tcatggaaaa gcttctgaat gtgtttgttt ggattggcta 2341 tgtttgttca ggaatcaatc ctctggtgta tactctgttc aacaaaattt accgaagggc 2401 attctccaac tatttgcgtt gcaattataa ggtagagaaa aagcctcctg tcaggcagat 2461 tccaagagtt gccgccactg ctttgtctgg gagggagctt aatgttaaca tttatcggca 2521 taccaatgaa ccggtgatcg agaaagccag tgacaatgag cccggtatag agatgcaagt 2581 tgagaattta gagttaccag taaatccctc cagtgtggtt agcgaaagga ttagcagtgt 2641 gtgaaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat 2701 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca 2761 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc 2821 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc 2881 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt 2941 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg 3001 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc 3061 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta 3121 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg 3181 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc 3241 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc 3301 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata 3361 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg 3421 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat 3481 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg 3541 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc 3601 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg 3661 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta 3721 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga 3781 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct 3841 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct 3901 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta 3961 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag 4021 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 4081 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 4141 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 4201 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata 4261 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 4321 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc 4381 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa 4441 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc 4501 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 4561 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 4621 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 4681 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa 4741 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 4801 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 4861 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 4921 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 4981 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 5041 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 5101 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 5161 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 5221 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 5281 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 5341 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 5401 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 5461 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 5521 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 5581 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 5641 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 5701 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 5761 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 5821 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 5881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 5941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6001 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt 6061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6121 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6181 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6241 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6301 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6361 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6421 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6481 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt //