LOCUS       sequence-412184-        7590 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the red 5-HT sensor GRAB_r5-HT1.0 in neurons in the 
            presence of Cre recombinase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7590)
  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 7590)
  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_r5-HT1.0
FEATURES             Location/Qualifiers
     source          1..7590
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        101..1279
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          332..1270
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1227..1247
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1310..1343)
                     /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(1394..1427)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(1947..1966)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(3345..3369)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     sig_peptide     complement(3373..3432)
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     regulatory      3429..3438
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    3445..3478
                     /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    3529..3562
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3587..4175
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3640..3660)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4058..4069)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4207..4683
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4406..4425)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4723..4863
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4723..4852
                     /label=AAV2 ITR
     rep_origin      4938..5393
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5025..5044)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5235..5256
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5410..5429)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5529..5551
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5589..5607)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5675..5779
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5780..5848,5849..6640)
                     /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(5998..6017)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6811..7399
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7300..7319
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   7461..7590
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga ggaatctttg
       61 cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg
      121 ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg gcaattgaac
      181 cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg
      241 cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg tgaacgttct
      301 ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt cccgcgggcc
      361 tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct ggctgcagta
      421 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc
      481 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg
      541 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat
      601 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc
      661 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg
      721 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg
      781 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat
      841 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg
      901 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg
      961 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg
     1021 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag
     1081 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg
     1141 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct
     1201 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct
     1261 tccatttcag gtgtcgtgag gtaccggatc ctctagagtc gactccggaa taacttcgta
     1321 taggatactt tatacgaagt tatgcagaat ggtagctgga ttgtagctgc tattagcaat
     1381 atgaaacctc ttaataactt cgtatagcat acattatacg aagttatggc gcatatgtca
     1441 ttacacactg ctaatccttt cgctaaccac actggaggga tttactggta actctaaatt
     1501 ctcaacttgc atctctatac cgggctcatt gtcactggct ttctcgatca ccggttcatt
     1561 ggtatgccga taaatgttaa cattaagctc cctcccagac aaagcagtgg cggcaactct
     1621 tggaatctgc ctgacaggag gctttttctc taccttataa ttgcaacgga ggatgatgag
     1681 gaaggcacgt ctgaaagact tattcaagaa ggcatagaga aaagggttca accccgaatt
     1741 gatatagcca agccagagga aagcgatcca cacctgctcg gggacagtgt agtctatgaa
     1801 agggtccaca atattggtga caaagaaggg ggcccagcag aaacagaagc agcccatgat
     1861 gatacataaa gtcttggctg ccttggtctc tgtcctcatg cgacttgtgc tgtgctggtc
     1921 gtgcaccgtc agctcccagc ccatggtctt cttctgcatt acggggccgt cgggggggaa
     1981 gttggtgccg cgcagcttca ccttgtagat gaagtagccg tcctgcaggg aggagtcctg
     2041 cagaacgtga atgcggccgc cgtcctcgaa gttcatcacg cgctcccacc tgaagccctc
     2101 ggggaaggac agcttgaagt agtcggggat gtcggctggg tgcttaatgt aggccttgga
     2161 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     2221 caccttcagc ttagcggtct gaaaggcctc gtaggggcgg ccctcgccct cgccctcgat
     2281 ctcgaactcg tggccgttga aggagccctc catgtgcacc ttgaagcgca tgaactcctt
     2341 gatgatggcg tggttgtgct cctcgccctt gctcaccaga ctcccgcctg tacctccctt
     2401 gtacagctcg tccatgccgc cggtggagtg gcgggcctcg gcgcgttcgc actgttccac
     2461 gatggtgtag tcctcgttgt gggacacgat gtccaacttg atgtcgacga tgtaggcgcc
     2521 gggcagctgc acgggcttct tggccttgta ggtggtcttg acctcggcgg cgtagtggcc
     2581 gccgtccttc agcctcagcc ccttcacgat ctcgctcttc agggcgccgt cctcggggta
     2641 cataacctcg gaacacacgg gccagagctg ggcatgctcc ttagcagtga catagattcg
     2701 gtaataggcc agcaccatga ggagaaacgg gatgtagaag gccaccacag agcaggtgat
     2761 agcatagggc ttgttgacca tgaagacaca ccacgtggag ttagagttgt ggctgaattt
     2821 ccttttctct atcacatcaa ctatgccgat gttgttccag ccttgcatta tggggagaaa
     2881 agatataaac atgggaagga cccagcagcc tcccaacatt aatgcgatgc gtagaggggt
     2941 catcttgttc ctataaacca aaggctggca gcagatggcg taatacctgt ccagggaaat
     3001 acagcacagg tgaaagatcg atgctgtaca aaggtagaca tccagagagg tccggaccag
     3061 gcagaacatc tccccataag cccagatgtc ttggaccagc tcaatggcac caaagggcat
     3121 caccagcacc gaaaccagca ggtcagcaaa ggcgagagac acaatgaaat agttggtttt
     3181 tattttcctg agctgcctgt ccctgcacac agccaccatc accagcaggt tgcccaagat
     3241 ggccatcagg ataaccactg caaggaacgt gagcagcacg accttctcca cggacctgaa
     3301 accctcgttg gaactcacat tagcatcaag tttgtcaccg gtggtgccaa cttttttgta
     3361 caaacttgtg tcaccagtgg aacctggaac ccagagcagc agtacccata gcaggagtgt
     3421 gtctgtctcc atggtggcgc tagcataact tcgtataaag tatcctatac gaagttattt
     3481 gccttaaccc agaaattatc actgttattc tttagaatgg tgcaaagaat aacttcgtat
     3541 aatgtatgct atacgaagtt atgaattcga tatcaagctt atcgataatc aacctctgga
     3601 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3661 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3721 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3781 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3841 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3901 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3961 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct atgttgccac
     4021 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     4081 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     4141 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag
     4201 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
     4261 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
     4321 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
     4381 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
     4441 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
     4501 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
     4561 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
     4621 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
     4681 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg
     4741 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg
     4801 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc
     4861 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac
     4921 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     4981 tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt tcgctttctt
     5041 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     5101 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga
     5161 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     5221 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact ctatctcggg
     5281 ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa aaaatgagct
     5341 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg
     5401 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac
     5461 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     5521 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     5581 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     5641 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5701 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5761 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5821 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5881 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5941 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     6001 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     6061 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     6121 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     6181 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     6241 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     6301 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     6361 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     6421 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     6481 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     6541 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     6601 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6661 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6721 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6781 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     6841 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     6901 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     6961 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     7021 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     7081 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     7141 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     7201 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     7261 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     7321 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     7381 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     7441 ctggcctttt gctcacatgt cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc
     7501 ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg
     7561 gagtggccaa ctccatcact aggggttcct
//
