LOCUS       sequence-412169-        7255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the green 5-HT sensor GRAB_g5-HT3.0 in astrocytes.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7255)
  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 7255)
  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-GfaABC1D-GRAB_g5-HT3.0
FEATURES             Location/Qualifiers
     source          1..7255
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   16..145
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     intron          1010..1142
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1125..1144
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1187..1205
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1222..1231
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     sig_peptide     1228..1287
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     protein_bind    1291..1315
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     2379..2400
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2506..2527)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2767..2786)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    3397..3985
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3450..3470)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3868..3879)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4017..4493
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4216..4235)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4533..4673
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4533..4662
                     /label=AAV2 ITR
     rep_origin      4748..5203
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4835..4854)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5045..5066
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5220..5239)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5339..5361
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5399..5417)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5485..5589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5590..5658,5659..6450)
                     /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"
     rep_origin      6621..7209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7110..7129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg
       61 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
      121 gccaactcca tcactagggg ttcctgcggc cgcacgcgtt gtagttaatg attaacccgc
      181 catgctactt atctacgtag ccatgctcta ggaagatcta acatatcctg gtgtggagta
      241 ggggacgctg ctctgacaga ggctcggggg cctgagctgg ctctgtgagc tggggaggag
      301 gcagacagcc aggccttgtc tgcaagcaga cctggcagca ttgggctggc cgccccccag
      361 ggcctcctct tcatgcccag tgaatgactc accttggcac agacacaatg ttcggggtgg
      421 gcacagtgcc tgcttcccgc cgcaccccag cccccctcaa atgccttccg agaagcccat
      481 tgagcagggg gcttgcattg caccccagcc tgacagcctg gcatcttggg ataaaagcag
      541 cacagccccc taggggctgc ccttgctgtg tggcgccacc ggcggtggag aacaaggctc
      601 tattcagcct gtgcccagga aaggggatca ggggatgccc aggcatggac agtgggtggc
      661 agggggggag aggagggctg tctgcttccc agaagtccaa ggacacaaat gggtgagggg
      721 agagctctcc ccatagctgg gctgcggccc aaccccaccc cctcaggcta tgccaggggg
      781 tgttgccagg ggcacccggg catcgccagt ctagcccact ccttcataaa gccctcgcat
      841 cccaggagcg agcagagcca gagcaggttg gagaggagac gcatcacctc cgctgctcgc
      901 aagctttatt gcggtagttt atcacagtta aattgctaac gcagtcagtg cttctgacac
      961 aacagtctcg aacttaagct gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
     1021 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
     1081 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1141 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1201 ataggctagc ctcgagaatt cgccaccatg gagacagaca cactcctgct atgggtactg
     1261 ctgctctggg ttccaggttc cactggtgac acaagtttgt acaaaaaagt tggcaccacc
     1321 ggtgacaaac ttgatgctaa tgtgagttcc aacgagggtt tcaggtccgt ggagaaggtc
     1381 gtgctgctca cgttccttgc agtggttatc ctgatggcca tcttgggcaa cctgctggtg
     1441 atggtggctg tgtgcaggga caggcagctc aggaaaataa aaaccaacta tttcattgtg
     1501 tctctcgcct ttgctgacct gctggtttcg gtgctggtga tgccctttgg tgccattgag
     1561 ctggtccaag acatctgggc ttatggggag atgttctgcc tggtccggac ctctctggat
     1621 gtctaccttt gtacagcatc gatctttcac ctgtgctgta tttccctgga caggtattac
     1681 gccatctgct gccagccttt ggtttatagg aacaagatga cccctctacg catcgcatta
     1741 atgttgggag gctgctgggt ccttcccatg tttatatctt ttctccccat aatgcaaggc
     1801 tggaacaaca tcggcatagt tgatgtgata gagaaaagga aattcagcca caactctaac
     1861 tccacgtggt gtgtcttcat ggtcaacaag ccctatgcta tcacctgctc tgtggtggcc
     1921 ttctacatcc cgtttctcct catggtgctg gcctattacc gaatctatgt cgtggctaag
     1981 cggggtccgg acgccgtcgc cgcgccgccg gggggcaccg agcgcaggcc caacggtctg
     2041 ggccccgagc gcagcgcggg cccggggggc gcagaggccg aaccgctgcc cacccagctc
     2101 aacggcgccc ctggcgagcc cgcgccggcc gggccgcgcg acaccgacgc gctggacctg
     2161 gaggagggag gaaacgtcta tatcaaggcc gacaagcaga agaacggcat caaggcgatc
     2221 ttccacatcc gccacaacat cgaggacggc ggcgtgcagc tcgccatcca ctaccagcag
     2281 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcgtgcag
     2341 tccaaacttt cgaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     2401 accgccgccg ggatcactct cggcatggac gagctgtaca agggcggtac cggagggagc
     2461 atggtgagaa agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     2521 ggcgacgtaa acggccacaa gttcagcgtg cggggcgagg gtgagggcga tgccaccaat
     2581 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2641 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagtc gctaccccga ccacatgaag
     2701 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tgcaggagcg caccatcttc
     2761 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2821 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2881 aagctggagt acaacaccgg agcagcagca cgcttccgct ggaggacaga gaccaaggca
     2941 gccaaggtgt tatgtgtcat catgggctgc ttctgtttct gctgggcccc cttctttgtc
     3001 accaatattg tggacccttt catagactac actgtccccg agcaggtgtg gactgctttc
     3061 ctctggcttg gctatatcaa ttcggggttg aacccttttc tctatgcctt cttgaataag
     3121 tctttcagac gtgccttcct catcatcctc cgttgcaatt ataaggtaga gaaaaagcct
     3181 cctgtcaggc agattccaag agttgccgcc actgctttgt ctgggaggga gcttaatgtt
     3241 aacatttatc ggcataccaa tgaaccggtg atcgagaaag ccagtgacaa tgagcccggt
     3301 atagagatgc aagttgagaa tttagagtta ccagtaaatc cctccagtgt ggttagcgaa
     3361 aggattagca gtgtgtgaga tatcaagctt atcgataatc aacctctgga ttacaaaatt
     3421 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     3481 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
     3541 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
     3601 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
     3661 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
     3721 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
     3781 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac ctggattctg
     3841 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
     3901 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
     3961 atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag atctacgggt
     4021 ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc actccagtgc
     4081 ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag gtgtccttct
     4141 ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg aagacaacct
     4201 gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa tcttggctca
     4261 ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc gagttgttgg
     4321 gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag agacggggtt
     4381 tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac ccaccttggc
     4441 ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc cttctgattt
     4501 tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg agttggccac
     4561 tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc
     4621 gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct
     4681 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa
     4741 ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     4801 gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt
     4861 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     4921 cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt
     4981 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     5041 aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt
     5101 gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa
     5161 aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt
     5221 acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     5281 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     5341 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     5401 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5461 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5521 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5581 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5641 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5701 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5761 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5821 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5881 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5941 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     6001 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     6061 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     6121 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6181 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6241 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6301 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6361 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6421 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6481 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6541 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6601 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6661 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6721 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6781 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6841 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6901 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6961 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7021 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7081 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7141 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7201 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggc
//
