LOCUS       sequence-412598-        6914 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the green 5-HT sensor GRAB_g5-HT2h in neurons in the 
            presence of Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6914)
  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 6914)
  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-DIO-GRAB_g5-HT2h
FEATURES             Location/Qualifiers
     source          1..6914
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   12..141
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     protein_bind    complement(616..649)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(700..733)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     1339..1358
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1598..1619
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1725..1746)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2810..2834)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(2838..2897)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      2894..2903
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2910..2943
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2994..3027
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3052..3640
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3105..3125)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3523..3534)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3672..4148
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3871..3890)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4188..4328
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4188..4317
                     /label=AAV2 ITR
     rep_origin      4403..4858
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4490..4509)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4700..4721
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4875..4894)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4994..5016
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5054..5072)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5140..5244
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5245..5313,5314..6105)
                     /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(5463..5482)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6276..6864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6765..6784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgctcacatg tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtatgag gcggggtggg ggtgcctacc
      181 tgacgaccga ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc
      241 ccctatcaga gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt
      301 cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca
      361 ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg
      421 gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg
      481 ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg
      541 tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc gagaaaccgg tgccacctct
      601 agagtcgact ccggaataac ttcgtatagg atactttata cgaagttatg cagaatggta
      661 gctggattgt agctgctatt agcaatatga aacctcttaa taacttcgta tagcatacat
      721 tatacgaagt tatggcgcat atgtcattac acactgctaa tcctttcgct aaccacactg
      781 gagggattta ctggtaactc taaattctca acttgcatct ctataccggg ctcattgtca
      841 ctggctttct cgatcaccgg ttcattggta tgccgataaa tgttaacatt aagctccctc
      901 ccagacaaag cagtggcggc aactcttgga atctgcctga caggaggctt tttctctacc
      961 ttataattgc aacggaggat gatgaggaag gcacgtctga aagacttatt caagaaggca
     1021 tagagaaaag ggttcaaccc cgaattgata tagccaagcc agaggaaagc agtccacacc
     1081 tgctcgggga cagtgtagtc tatgaaaggg tccacaatat tggtgacaaa gaagggggcc
     1141 cagcagaaac agaagcagcc catgatgaca cataaagtct tggctgcctt ggtctctgtc
     1201 ctcatgcgcc agcgtgctgc tgctccggtg ttgtactcca gcttgtgccc caggatgttg
     1261 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     1321 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     1381 tggatgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     1441 tagcgactga agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg
     1501 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccctcggt ggcatcgccc
     1561 tcaccctcgc cccgcacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     1621 atgggcacca ccccggtgaa cagctcctcg ccctttctca ccatgctccc tccggtaccg
     1681 cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     1741 accatgtgat cgcgcttctc gttggggtct ttcgaaagtt tggactgcac gctcaggtag
     1801 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggatg
     1861 gcgagctgca cgccgccgtc ctcgatgttg tggcggatgt ggaagatcgc cttgatgccg
     1921 ttcttctgct tgtcggcctt gatatagacg tttcctccct cctccaggtc cagcgcgtcg
     1981 gtgtcgcgcg gcccggccgg cgcgggctcg ccaggggcgc cgttgagctg ggtgggcagc
     2041 ggttcggcct ctgcgccccc cgggcccgcg ctgcgctcgg ggcccagacc gttgggcctg
     2101 cgctcggtgc cccccggcgg cgcggcgacg gcgtccggac cccgcttagc agtgacatag
     2161 attcggtaat aggccagcac catgaggaga aacgggatgt agaaggccac cacagagcag
     2221 gtgatagcat agggcttgtt gaccatgaag acacaccacg tggagttaga gttgtggctg
     2281 aatttccttt tctctatcac atcaactatg ccgatgttgt tccagccttg cattatgggg
     2341 agaaaagata taaacatggg aaggacccag cagcctccca acattaatgc gatgcgtaga
     2401 ggggtcatct tgttcctata aaccaaaggc tggcagcaga tggcgtaata cctgtccagg
     2461 gaaatacagc acaggtgaaa gatcgatgct gtggtaagta ggacatccag agaggtccgg
     2521 accaggcaga acatctcccc ataagcccag atgtcttgga ccagctcaat ggcaccaaag
     2581 ggcatcacca gcaccgaaac cagcaggtca gcaaaggcga gagacacaat gaaatagttg
     2641 gtttttattt tcctgagctg cctgtccctg cacacagcca ccatcaccag caggttgccc
     2701 aagatggcca tcaggataac cactgcaagg aacgtgagca gcacgacctt ctccacggac
     2761 ctgaaaccct cgttggaact cacattagca tcaagtttgt caccggtggt gccaactttt
     2821 ttgtacaaac ttgtgtcacc agtggaacct ggaacccaga gcagcagtac ccatagcagg
     2881 agtgtgtctg tctccatggt ggcgctagca taacttcgta taaagtatcc tatacgaagt
     2941 tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt
     3001 cgtataatgt atgctatacg aagttatgaa ttcgatatca agcttatcga taatcaacct
     3061 ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg
     3121 ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc
     3181 attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt
     3241 gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc
     3301 attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg
     3361 gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact
     3421 gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt
     3481 gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg
     3541 gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc
     3601 cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc
     3661 gagagatcta cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag
     3721 ttgccactcc agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg
     3781 actaggtgtc cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag
     3841 ttgggaagac aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg
     3901 cacaatcttg gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc
     3961 ctcccgagtt gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt
     4021 ggtagagacg gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga
     4081 tctacccacc ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc
     4141 ctgtccttct gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt
     4201 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     4261 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     4321 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     4381 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     4441 gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc tcctttcgct
     4501 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     4561 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     4621 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     4681 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caactctatc
     4741 tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg gttaaaaaat
     4801 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     4861 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4921 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4981 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     5041 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5101 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     5161 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5221 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5281 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5341 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5401 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5461 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5521 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5581 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5641 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5701 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5761 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5821 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5881 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5941 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6001 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6061 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6121 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6181 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6241 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6301 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6361 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6421 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6481 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6541 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6601 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6661 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6721 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6781 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6841 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6901 ttttgctggc cttt
//
