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
//
