LOCUS       sequence-412608-        6635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the green 5-HT sensor GRAB_g5-HT2h 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-HT2h
FEATURES             Location/Qualifiers
     source          1..6635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     68..87
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   229..358
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     regulatory      809..818
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     815..874
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    878..902
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     1966..1987
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2093..2114)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2354..2373)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2990..3578
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3043..3063)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3461..3472)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3610..4086
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3809..3828)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4126..4266
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4126..4255
                     /label=AAV2 ITR
     rep_origin      4341..4796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4428..4447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4638..4659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4813..4832)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4932..4954
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4992..5010)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5078..5182
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5183..5251,5252..6043)
                     /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(5401..5420)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6214..6635,1..167)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
       61 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
      121 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
      181 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc
      241 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc
      301 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc
      361 ggccgcacgc gtatgaggcg gggtgggggt gcctacctga cgaccgaccc cgacccactg
      421 gacaagcacc caacccccat tccccaaatt gcgcatcccc tatcagagag ggggagggga
      481 aacaggatgc ggcgaggcgc gtgcgcactg ccagcttcag caccgcggac agtgccttcg
      541 cccccgcctg gcggcgcgcg ccaccgccgc ctcagcactg aaggcgcgct gacgtcactc
      601 gccggtcccc cgcaaactcc ccttcccggc caccttggtc gcgtccgcgc cgccgccggc
      661 ccagccggac cgcaccacgc gaggcgcgag ataggggggc acgggcgcga ccatctgcgc
      721 tgcggcgccg gcgactcagc gctgcctcag tctgcggtgg gcagcggagg agtcgtgtcg
      781 tgcctgagag cgcagtcgag aaaccggtgc caccatggag acagacacac tcctgctatg
      841 ggtactgctg ctctgggttc caggttccac tggtgacaca agtttgtaca aaaaagttgg
      901 caccaccggt gacaaacttg atgctaatgt gagttccaac gagggtttca ggtccgtgga
      961 gaaggtcgtg ctgctcacgt tccttgcagt ggttatcctg atggccatct tgggcaacct
     1021 gctggtgatg gtggctgtgt gcagggacag gcagctcagg aaaataaaaa ccaactattt
     1081 cattgtgtct ctcgcctttg ctgacctgct ggtttcggtg ctggtgatgc cctttggtgc
     1141 cattgagctg gtccaagaca tctgggctta tggggagatg ttctgcctgg tccggacctc
     1201 tctggatgtc ctacttacca cagcatcgat ctttcacctg tgctgtattt ccctggacag
     1261 gtattacgcc atctgctgcc agcctttggt ttataggaac aagatgaccc ctctacgcat
     1321 cgcattaatg ttgggaggct gctgggtcct tcccatgttt atatcttttc tccccataat
     1381 gcaaggctgg aacaacatcg gcatagttga tgtgatagag aaaaggaaat tcagccacaa
     1441 ctctaactcc acgtggtgtg tcttcatggt caacaagccc tatgctatca cctgctctgt
     1501 ggtggccttc tacatcccgt ttctcctcat ggtgctggcc tattaccgaa tctatgtcac
     1561 tgctaagcgg ggtccggacg ccgtcgccgc gccgccgggg ggcaccgagc gcaggcccaa
     1621 cggtctgggc cccgagcgca gcgcgggccc ggggggcgca gaggccgaac cgctgcccac
     1681 ccagctcaac ggcgcccctg gcgagcccgc gccggccggg ccgcgcgaca ccgacgcgct
     1741 ggacctggag gagggaggaa acgtctatat caaggccgac aagcagaaga acggcatcaa
     1801 ggcgatcttc cacatccgcc acaacatcga ggacggcggc gtgcagctcg ccatccacta
     1861 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1921 cgtgcagtcc aaactttcga aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1981 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagg gcggtaccgg
     2041 agggagcatg gtgagaaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     2101 gctggacggc gacgtaaacg gccacaagtt cagcgtgcgg ggcgagggtg agggcgatgc
     2161 caccgagggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     2221 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagtcgct accccgacca
     2281 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacatcc aggagcgcac
     2341 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     2401 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     2461 ggggcacaag ctggagtaca acaccggagc agcagcacgc tggcgcatga ggacagagac
     2521 caaggcagcc aagactttat gtgtcatcat gggctgcttc tgtttctgct gggccccctt
     2581 ctttgtcacc aatattgtgg accctttcat agactacact gtccccgagc aggtgtggac
     2641 tgctttcctc tggcttggct atatcaattc ggggttgaac ccttttctct atgccttctt
     2701 gaataagtct ttcagacgtg ccttcctcat catcctccgt tgcaattata aggtagagaa
     2761 aaagcctcct gtcaggcaga ttccaagagt tgccgccact gctttgtctg ggagggagct
     2821 taatgttaac atttatcggc ataccaatga accggtgatc gagaaagcca gtgacaatga
     2881 gcccggtata gagatgcaag ttgagaattt agagttacca gtaaatccct ccagtgtggt
     2941 tagcgaaagg attagcagtg tgtaagaatt cgatatcaag cttatcgata atcaacctct
     3001 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     3061 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     3121 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     3181 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     3241 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     3301 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     3361 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     3421 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     3481 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     3541 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     3601 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3661 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3721 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3781 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     3841 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     3901 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     3961 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     4021 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     4081 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     4141 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4201 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4261 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4321 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     4441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4681 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4741 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4801 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4861 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4921 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4981 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5041 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5101 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5161 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5221 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5281 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5341 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5401 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5461 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5521 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5581 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5641 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5701 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5761 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5821 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5881 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5941 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6001 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6061 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6121 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6181 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6241 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6301 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6361 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6601 tgagctatga gaaagcgcca cgcttcccga aggga
//
