LOCUS       sequence-412607-        7749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the green 5-HT sensor GRAB_g5-HT3.0 in neurons in the 
            presence of Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7749)
  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 7749)
  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-EF1a-DIO-GRAB_g5-HT3.0
FEATURES             Location/Qualifiers
     source          1..7749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     61..80
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   222..351
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        452..1630
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          683..1621
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1578..1598
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1661..1694)
                     /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(1745..1778)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2384..2403
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2643..2664
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2770..2791)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(3855..3879)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(3883..3942)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3939..3948
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3955..3988
                     /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    4039..4072
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4097..4685
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4150..4170)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4568..4579)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4717..5193
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4916..4935)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5233..5373
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5233..5362
                     /label=AAV2 ITR
     rep_origin      5448..5903
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5535..5554)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5745..5766
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5920..5939)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6039..6061
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6099..6117)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6185..6289
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6290..6358,6359..7150)
                     /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(6508..6527)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7321..7749,1..160)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
       61 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
      121 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
      181 tttacggttc ctggcctttt gctggccttt tgctcacatg tcctgcaggc agctgcgcgc
      241 tcgctcgctc actgaggccg cccgggcgtc gggcgacctt tggtcgcccg gcctcagtga
      301 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc tgcggccgca
      361 cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag aggaatcttt gcagctaatg
      421 gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt gggcagagcg
      481 cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa ccggtgccta
      541 gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc
      601 cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa
      661 cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc ctggcctctt
      721 tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt acgtgattct
      781 tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg cttaaggagc
      841 cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc gcgtgcgaat
      901 ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca tttaaaattt
      961 ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg cgggccaaga
     1021 tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc gtgcgtccca
     1081 gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg gacgggggta
     1141 gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta tcgccccgcc
     1201 ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat ggccgcttcc
     1261 cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc gggcgggtga
     1321 gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat gtgactccac
     1381 ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga gtacgtcgtc
     1441 tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt gggtggagac
     1501 tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc tttttgagtt
     1561 tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc ttccatttca
     1621 ggtgtcgtga ggtaccggat cctctagagt cgactccgga ataacttcgt ataggatact
     1681 ttatacgaag ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct
     1741 cttaataact tcgtatagca tacattatac gaagttatgg cgcatatgtc attacacact
     1801 gctaatcctt tcgctaacca cactggaggg atttactggt aactctaaat tctcaacttg
     1861 catctctata ccgggctcat tgtcactggc tttctcgatc accggttcat tggtatgccg
     1921 ataaatgtta acattaagct ccctcccaga caaagcagtg gcggcaactc ttggaatctg
     1981 cctgacagga ggctttttct ctaccttata attgcaacgg aggatgatga ggaaggcacg
     2041 tctgaaagac ttattcaaga aggcatagag aaaagggttc aaccccgaat tgatatagcc
     2101 aagccagagg aaagcagtcc acacctgctc ggggacagtg tagtctatga aagggtccac
     2161 aatattggtg acaaagaagg gggcccagca gaaacagaag cagcccatga tgacacataa
     2221 caccttggct gccttggtct ctgtcctcca gcggaagcgt gctgctgctc cggtgttgta
     2281 ctccagcttg tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat
     2341 gcggttcacc agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc
     2401 gtccttgaag aagatggtgc gctcctgcac gtagccttcg ggcatggcgg acttgaagaa
     2461 gtcgtgctgc ttcatgtggt cggggtagcg actgaagcac tgcacgccgt aggtcagggt
     2521 ggtcacgagg gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt
     2581 cagcttgcca ttggtggcat cgccctcacc ctcgccccgc acgctgaact tgtggccgtt
     2641 tacgtcgccg tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt
     2701 tctcaccatg ctccctccgg taccgccctt gtacagctcg tccatgccga gagtgatccc
     2761 ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttcga
     2821 aagtttggac tgcacgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat
     2881 gggggtgttc tgctggtagt ggatggcgag ctgcacgccg ccgtcctcga tgttgtggcg
     2941 gatgtggaag atcgccttga tgccgttctt ctgcttgtcg gccttgatat agacgtttcc
     3001 tccctcctcc aggtccagcg cgtcggtgtc gcgcggcccg gccggcgcgg gctcgccagg
     3061 ggcgccgttg agctgggtgg gcagcggttc ggcctctgcg ccccccgggc ccgcgctgcg
     3121 ctcggggccc agaccgttgg gcctgcgctc ggtgcccccc ggcggcgcgg cgacggcgtc
     3181 cggaccccgc ttagccacga catagattcg gtaataggcc agcaccatga ggagaaacgg
     3241 gatgtagaag gccaccacag agcaggtgat agcatagggc ttgttgacca tgaagacaca
     3301 ccacgtggag ttagagttgt ggctgaattt ccttttctct atcacatcaa ctatgccgat
     3361 gttgttccag ccttgcatta tggggagaaa agatataaac atgggaagga cccagcagcc
     3421 tcccaacatt aatgcgatgc gtagaggggt catcttgttc ctataaacca aaggctggca
     3481 gcagatggcg taatacctgt ccagggaaat acagcacagg tgaaagatcg atgctgtaca
     3541 aaggtagaca tccagagagg tccggaccag gcagaacatc tccccataag cccagatgtc
     3601 ttggaccagc tcaatggcac caaagggcat caccagcacc gaaaccagca ggtcagcaaa
     3661 ggcgagagac acaatgaaat agttggtttt tattttcctg agctgcctgt ccctgcacac
     3721 agccaccatc accagcaggt tgcccaagat ggccatcagg ataaccactg caaggaacgt
     3781 gagcagcacg accttctcca cggacctgaa accctcgttg gaactcacat tagcatcaag
     3841 tttgtcaccg gtggtgccaa cttttttgta caaacttgtg tcaccagtgg aacctggaac
     3901 ccagagcagc agtacccata gcaggagtgt gtctgtctcc atggtggcgc tagcataact
     3961 tcgtataaag tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc
     4021 tttagaatgg tgcaaagaat aacttcgtat aatgtatgct atacgaagtt atgaattcga
     4081 tatcaagctt atcgataatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat
     4141 tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca
     4201 tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc
     4261 tctttatgag gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc
     4321 tgacgcaacc cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt
     4381 cgctttcccc ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg
     4441 gacaggggct cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga aatcatcgtc
     4501 ctttccttgg ctgctcgcct atgttgccac ctggattctg cgcgggacgt ccttctgcta
     4561 cgtcccttcg gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg
     4621 gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc
     4681 cccgcatcga taccgagcgc tgctcgagag atctacgggt ggcatccctg tgacccctcc
     4741 ccagtgcctc tcctggccct ggaagttgcc actccagtgc ccaccagcct tgtcctaata
     4801 aaattaagtt gcatcatttt gtctgactag gtgtccttct ataatattat ggggtggagg
     4861 ggggtggtat ggagcaaggg gcaagttggg aagacaacct gtagggcctg cggggtctat
     4921 tgggaaccaa gctggagtgc agtggcacaa tcttggctca ctgcaatctc cgcctcctgg
     4981 gttcaagcga ttctcctgcc tcagcctccc gagttgttgg gattccaggc atgcatgacc
     5041 aggctcagct aatttttgtt tttttggtag agacggggtt tcaccatatt ggccaggctg
     5101 gtctccaact cctaatctca ggtgatctac ccaccttggc ctcccaaatt gctgggatta
     5161 caggcgtgaa ccactgctcc cttccctgtc cttctgattt tgtaggtaac cacgtgcgga
     5221 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct
     5281 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct
     5341 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta
     5401 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     5461 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     5521 cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     5581 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     5641 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     5701 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     5761 aactggaaca acactcaact ctatctcggg ctattctttt gatttataag ggattttgcc
     5821 gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     5881 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     5941 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6001 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     6061 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     6121 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     6181 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     6241 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     6301 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     6361 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     6421 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     6481 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     6541 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     6601 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6661 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6721 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6781 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6841 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6901 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6961 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     7021 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     7081 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     7141 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     7201 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     7261 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     7321 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     7381 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     7441 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     7501 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     7561 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     7621 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7681 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7741 gagaaaggc
//
