LOCUS       sequence-412600-        6635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the green 5-HT sensor GRAB_g5-HT3.0mut in neurons.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6635)
  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 6635)
  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-hSyn-GRAB_g5-HT3.0mut
FEATURES             Location/Qualifiers
     source          1..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      581..590
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     587..646
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    650..674
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1738..1759
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1865..1886)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2126..2145)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2762..3350
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2815..2835)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3233..3244)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3382..3858
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3581..3600)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3898..4038
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3898..4027
                     /label=AAV2 ITR
     rep_origin      4113..4568
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4200..4219)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4410..4431
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4585..4604)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4704..4726
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4764..4782)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4850..4954
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4955..5023,5024..5815)
                     /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(5173..5192)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5986..6574
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6475..6494
                     /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 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 gtgacaaact tgatgctaat gtgagttcca acgagggttt
      721 caggtccgtg gagaaggtcg tgctgctcac gttccttgca gtggttatcc tgatggccat
      781 cttgggcaac ctgctggtga tggtggctgt gtgcagggac aggcagctca ggaaaataaa
      841 aaccaactat ttcattgtgt ctctcgcctt tgctgacctg ctggtttcgg tgctggtgat
      901 gccctttggt gccattgagc tggtccaaga catctgggct tatggggaga tgttctgcct
      961 ggtccggacc tctctgtttg tctacctttg tacagcatcg atctttcacc tgtgctgtat
     1021 ttccctggac aggtattacg ccatctgctg ccagcctttg gtttatagga acaagatgac
     1081 ccctctacgc atcgcattaa tgttgggagg ctgctgggtc cttcccatgt ttatatcttt
     1141 tctccccata atgcaaggct ggaacaacat cggcatagtt gatgtgatag agaaaaggaa
     1201 attcagccac aactctaact ccacgtggtg tgtcttcatg gtcaacaagc cctatgctat
     1261 cacctgctct gtggtggcct tctacatccc gtttctcctc atggtgctgg cctattaccg
     1321 aatctatgtc gtggctaagc ggggtccgga cgccgtcgcc gcgccgccgg ggggcaccga
     1381 gcgcaggccc aacggtctgg gccccgagcg cagcgcgggc ccggggggcg cagaggccga
     1441 accgctgccc acccagctca acggcgcccc tggcgagccc gcgccggccg ggccgcgcga
     1501 caccgacgcg ctggacctgg aggagggagg aaacgtctat atcaaggccg acaagcagaa
     1561 gaacggcatc aaggcgatct tccacatccg ccacaacatc gaggacggcg gcgtgcagct
     1621 cgccatccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     1681 ccactacctg agcgtgcagt ccaaactttc gaaagacccc aacgagaagc gcgatcacat
     1741 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     1801 gggcggtacc ggagggagca tggtgagaaa gggcgaggag ctgttcaccg gggtggtgcc
     1861 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgc ggggcgaggg
     1921 tgagggcgat gccaccaatg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1981 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagtcg
     2041 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2101 gcaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2161 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2221 cggcaacatc ctggggcaca agctggagta caacaccgga gcagcagcac gcttccgctg
     2281 gaggacagag accaaggcag ccaaggtgtt atgtgtcatc atgggctgct tctgtttctg
     2341 ctgggccccc ttctttgtca ccaatattgt ggaccctttc atagactaca ctgtccccga
     2401 gcaggtgtgg actgctttcc tctggcttgg ctatatcaat tcggggttga acccttttct
     2461 ctatgccttc ttgaataagt ctttcagacg tgccttcctc atcatcctcc gttgcaatta
     2521 taaggtagag aaaaagcctc ctgtcaggca gattccaaga gttgccgcca ctgctttgtc
     2581 tgggagggag cttaatgtta acatttatcg gcataccaat gaaccggtga tcgagaaagc
     2641 cagtgacaat gagcccggta tagagatgca agttgagaat ttagagttac cagtaaatcc
     2701 ctccagtgtg gttagcgaaa ggattagcag tgtgtaagaa ttcgatatca agcttatcga
     2761 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2821 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2881 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2941 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     3001 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     3061 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     3121 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3181 cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3241 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3301 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3361 agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg
     3421 gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc
     3481 attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc
     3541 aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg
     3601 agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc
     3661 ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt
     3721 ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa
     3781 tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact
     3841 gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg
     3901 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     3961 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     4021 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     4081 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     4141 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     4201 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     4261 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     4321 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     4381 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     4441 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     4501 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     4561 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     4621 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     4681 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     4741 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     4801 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     4861 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     4921 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     4981 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     5041 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     5101 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     5161 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     5221 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     5281 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     5341 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     5401 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     5461 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     5521 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     5581 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     5641 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     5701 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     5761 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     5821 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5881 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5941 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     6001 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     6061 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     6121 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     6181 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     6241 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6301 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6361 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6421 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6481 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6541 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6601 gttcctggcc ttttgctggc cttttgctca catgt
//
