LOCUS       sequence-412606-        7514 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the green 5-HT sensor GRAB_g5-HT3.0 in excitatory neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7514)
  AUTHORS   Deng F, Wan J, Li G, Dong H, Xia X, Wang Y, Li X, Zhuang C, Zheng Y,
            Liu L, Yan Y, Feng J, Zhao Y, Xie H, Li Y
  TITLE     Improved green and red GRAB sensors for monitoring spatiotemporal 
            serotonin release in vivo.
  JOURNAL   Nat Methods. 2024 Apr;21(4):692-702. doi: 
            10.1038/s41592-024-02188-8. Epub 2024 Mar 5.
  PUBMED    38443508
REFERENCE   2  (bases 1 to 7514)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41592-024-02188-8"; journalName: "Nat Methods"; date: 
            "2024-04"; volume: "21"; issue: "4"; pages: "692-702"
COMMENT     Sequence Label: pAAV-CaMKII-GRAB_g5-HT3.0
FEATURES             Location/Qualifiers
     source          1..7514
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        145..1433
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1460..1469
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1466..1525
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    1529..1553
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2617..2638
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2744..2765)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3005..3024)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3641..4229
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3694..3714)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4112..4123)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4261..4737
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4460..4479)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4777..4917
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4777..4906
                     /label=AAV2 ITR
     rep_origin      4992..5447
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5079..5098)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5289..5310
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5464..5483)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5583..5605
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5643..5661)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5729..5833
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5834..5902,5903..6694)
                     /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(6052..6071)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6865..7453
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7354..7373
                     /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 gcgtcattat ggccttaggt cacttcatct ccatggggtt
      181 cttcttctga ttttctagaa aatgagatgg gggtgcagag agcttcctca gtgacctgcc
      241 cagggtcaca tcagaaatgt cagagctaga acttgaactc agattactaa tcttaaattc
      301 catgccttgg gggcatgcaa gtacgatata cagaaggagt gaactcatta gggcagatga
      361 ccaatgagtt taggaaagaa gagtccaggg cagggtacat ctacaccacc cgcccagccc
      421 tgggtgagtc cagccacgtt cacctcatta tagttgcctc tctccagtcc taccttgacg
      481 ggaagcacaa gcagaaactg ggacaggagc cccaggagac caaatcttca tggtccctct
      541 gggaggatgg gtggggagag ctgtggcaga ggcctcagga ggggccctgc tgctcagtgg
      601 tgacagatag gggtgagaaa gcagacagag tcattccgtc agcattctgg gtctgtttgg
      661 tacttcttct cacgctaagg tggcggtgtg atatgcacaa tggctaaaaa gcagggagag
      721 ctggaaagaa acaaggacag agacagaggc caagtcaacc agaccaattc ccagaggaag
      781 caaagaaacc attacagaga ctacaagggg gaagggaagg agagatgaat tagcttcccc
      841 tgtaaacctt agaacccagc tgttgccagg gcaacggggc aatacctgtc tcttcagagg
      901 agatgaagtt gccagggtaa ctacatcctg tctttctcaa ggaccatccc agaatgtggc
      961 acccactagc cgttaccata gcaactgcct ctttgcccca cttaatccca tcccgtctgt
     1021 taaaagggcc ctatagttgg aggtggggga ggtaggaaga gcgatgatca cttgtggact
     1081 aagtttgttc gcatcccctt ctccaacccc ctcagtacat caccctgggg gaacagggtc
     1141 cacttgctcc tgggcccaca cagtcctgca gtattgtgta tataaggcca gggcaaagag
     1201 gagcaggttt taaagtgaaa ggcaggcagg tgttggggag gcagttaccg gggcaacggg
     1261 aacagggcgt ttcggaggtg gttgccatgg ggacctggat gctgacgaag gctcgcgagg
     1321 ctgtgagcag ccacagtgcc ctgctcagaa gccccaagct cgtcagtcaa gccggttctc
     1381 cgtttgcact caggagcacg ggcaggcgag tggcccctag ttctgggggc agcggggtac
     1441 cggatcctct agagtcgacg ccaccatgga gacagacaca ctcctgctat gggtactgct
     1501 gctctgggtt ccaggttcca ctggtgacac aagtttgtac aaaaaagttg gcaccaccgg
     1561 tgacaaactt gatgctaatg tgagttccaa cgagggtttc aggtccgtgg agaaggtcgt
     1621 gctgctcacg ttccttgcag tggttatcct gatggccatc ttgggcaacc tgctggtgat
     1681 ggtggctgtg tgcagggaca ggcagctcag gaaaataaaa accaactatt tcattgtgtc
     1741 tctcgccttt gctgacctgc tggtttcggt gctggtgatg ccctttggtg ccattgagct
     1801 ggtccaagac atctgggctt atggggagat gttctgcctg gtccggacct ctctggatgt
     1861 ctacctttgt acagcatcga tctttcacct gtgctgtatt tccctggaca ggtattacgc
     1921 catctgctgc cagcctttgg tttataggaa caagatgacc cctctacgca tcgcattaat
     1981 gttgggaggc tgctgggtcc ttcccatgtt tatatctttt ctccccataa tgcaaggctg
     2041 gaacaacatc ggcatagttg atgtgataga gaaaaggaaa ttcagccaca actctaactc
     2101 cacgtggtgt gtcttcatgg tcaacaagcc ctatgctatc acctgctctg tggtggcctt
     2161 ctacatcccg tttctcctca tggtgctggc ctattaccga atctatgtcg tggctaagcg
     2221 gggtccggac gccgtcgccg cgccgccggg gggcaccgag cgcaggccca acggtctggg
     2281 ccccgagcgc agcgcgggcc cggggggcgc agaggccgaa ccgctgccca cccagctcaa
     2341 cggcgcccct ggcgagcccg cgccggccgg gccgcgcgac accgacgcgc tggacctgga
     2401 ggagggagga aacgtctata tcaaggccga caagcagaag aacggcatca aggcgatctt
     2461 ccacatccgc cacaacatcg aggacggcgg cgtgcagctc gccatccact accagcagaa
     2521 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcgtgcagtc
     2581 caaactttcg aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     2641 cgccgccggg atcactctcg gcatggacga gctgtacaag ggcggtaccg gagggagcat
     2701 ggtgagaaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     2761 cgacgtaaac ggccacaagt tcagcgtgcg gggcgagggt gagggcgatg ccaccaatgg
     2821 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     2881 cgtgaccacc ctgacctacg gcgtgcagtg cttcagtcgc taccccgacc acatgaagca
     2941 gcacgacttc ttcaagtccg ccatgcccga aggctacgtg caggagcgca ccatcttctt
     3001 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     3061 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     3121 gctggagtac aacaccggag cagcagcacg cttccgctgg aggacagaga ccaaggcagc
     3181 caaggtgtta tgtgtcatca tgggctgctt ctgtttctgc tgggccccct tctttgtcac
     3241 caatattgtg gaccctttca tagactacac tgtccccgag caggtgtgga ctgctttcct
     3301 ctggcttggc tatatcaatt cggggttgaa cccttttctc tatgccttct tgaataagtc
     3361 tttcagacgt gccttcctca tcatcctccg ttgcaattat aaggtagaga aaaagcctcc
     3421 tgtcaggcag attccaagag ttgccgccac tgctttgtct gggagggagc ttaatgttaa
     3481 catttatcgg cataccaatg aaccggtgat cgagaaagcc agtgacaatg agcccggtat
     3541 agagatgcaa gttgagaatt tagagttacc agtaaatccc tccagtgtgg ttagcgaaag
     3601 gattagcagt gtgtgagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa
     3661 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
     3721 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
     3781 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
     3841 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
     3901 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
     3961 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
     4021 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctatgttg ccacctggat
     4081 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
     4141 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
     4201 tcggatctcc ctttgggccg cctccccgca tcgataccga gcgctgctcg agagatctac
     4261 gggtggcatc cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca
     4321 gtgcccacca gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc
     4381 ttctataata ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca
     4441 acctgtaggg cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg
     4501 ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg
     4561 ttgggattcc aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg
     4621 ggtttcacca tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct
     4681 tggcctccca aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg
     4741 attttgtagg taaccacgtg cggaccgagc ggccgcagga acccctagtg atggagttgg
     4801 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4861 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4921 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4981 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5041 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     5101 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5161 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     5221 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5281 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     5341 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     5401 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     5461 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5521 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5581 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5641 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5701 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5761 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5821 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5881 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5941 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     6001 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     6061 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     6121 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     6181 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6241 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6301 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6361 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6421 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6481 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6541 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6601 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6661 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6721 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6781 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6841 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6901 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6961 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     7021 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     7081 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     7141 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     7201 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7261 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7321 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7381 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7441 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7501 ttttgctcac atgt
//
