LOCUS       sequence-447481-        6489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cilia-targeted serotonin sensor based on HTR6 receptor and
            cpEGFP, driven by SYN promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6489)
  AUTHORS   Sheu SH, Upadhyayula S, Dupuy V, Pang S, Deng F, Wan J, Walpita D, 
            Pasolli HA, Houser J, Sanchez-Martinez S, Brauchi SE, Banala S, 
            Freeman M, Xu CS, Kirchhausen T, Hess HF, Lavis L, Li Y, 
            Chaumont-Dubel S, Clapham DE
  TITLE     A serotonergic axon-cilium synapse drives nuclear signaling to alter
            chromatin accessibility.
  JOURNAL   Cell. 2022 Sep 1;185(18):3390-3407.e18. doi: 
            10.1016/j.cell.2022.07.026.
  PUBMED    36055200
REFERENCE   2  (bases 1 to 6489)
  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: 
            "10.1016/j.cell.2022.07"; journalName: "Cell"; date: "2022-09-1- 1";
            volume: "185"; issue: "18"; pages: "3390-3407.e18"
COMMENT     Sequence Label: pAAV-Syn1-GRAB-HTR6-cilia
FEATURES             Location/Qualifiers
     source          1..6489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    148..168
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        190..637
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    637..661
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      662..671
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1762..1783
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1889..1910)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2150..2169)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(2879..2903)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2913..3501
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2966..2986)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3384..3395)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3531..3548)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3537..3744
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3752..3892
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3752..3881
                     /label=AAV2 ITR
     rep_origin      3967..4422
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4054..4073)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4264..4285
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4439..4458)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4558..4580
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4618..4636)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4704..4808
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4809..4877,4878..5669)
                     /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(5027..5046)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5840..6428
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6329..6348
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttctagacaa ctttgtatag aaaagttgct gcagagggcc
      181 ctgcgtatga gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct
      241 gacgaccgac cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc
      301 cctatcagag agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc
      361 agcaccgcgg acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac
      421 tgaaggcgcg ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg
      481 tcgcgtccgc gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg
      541 gcacgggcgc gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt
      601 gggcagcgga ggagtcgtgt cgtgcctgag agcgcagcaa gtttgtacaa aaaagcaggc
      661 tgccaccatg gtgcccgagc cgggcccaac tgctaacagc accccagctt ggggggctgg
      721 tcctccaagc gccccaggtg gcagtggttg ggttgctgca gcattgtgcg tggttatcgc
      781 cctgacagct gccgcaaaca gcctgctcat tgctcttatt tgtacccaac ccgcactccg
      841 gaatacgtct aattttttcc tggtctcact gttcacgagc gaccttatgg tcgggctcgt
      901 agtcatgcct cctgctatgt tgaacgccct gtatgggaga tgggtcctgg caaggggtct
      961 ttgccttctg tggaccgcat ttgacgtcat gtgctgtagt gccagcatct tgaacctctg
     1021 tttgattagt ctggatagat atctgctgat tctgagccca cttagataca agctgcggat
     1081 gactccgctc cgggccctcg ctttggtgct gggcgcctgg tctctggcag cactggcttc
     1141 attcttgcct ttgctgctgg gctggcacga attggggcac gcaaggccgc cagttccagg
     1201 ccagtgtagg ctgcttgcct ccctgccctt cgtgttggtg gcttccggcc tcacattctt
     1261 tctgcctagt ggtgccatct gcttcactta ctgccggata cttctggcag ctagaaaaca
     1321 agccgttcag gttgcctccc tcaccaccgg catggcaagc cagcggggtc cggacgccgt
     1381 cgccgcgccg ccggggggca ccgagcgcag gcccaacggt ctgggccccg agcgcagcgc
     1441 gggcccgggg ggcgcagagg ccgaaccgct gcccacccag ctcaacggcg cccctggcga
     1501 gcccgcgccg gccgggccgc gcgacaccga cgcgctggac ctggaggagg gaggaaacgt
     1561 ctatatcaag gccgacaagc agaagaacgg catcaaggcg aacttccaca tccgccacaa
     1621 catcgaggac ggcggcgtgc agctcgccta ccactaccag cagaacaccc ccatcggcga
     1681 cggccccgtg ctgctgcccg acaaccacta cctgagcgtg cagtccaaac tttcgaaaga
     1741 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
     1801 tctcggcatg gacgagctgt acaagggcgg taccggaggg agcatggtga gaaagggcga
     1861 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     1921 caagttcagc gtgtccggcg agggtgaggg cgatgccacc gagggcaagc tgaccctgaa
     1981 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2041 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2101 gtccgccatg cccgaaggct acatccagga gcgcaccatc ttcttcaagg acgacggcaa
     2161 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2221 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacac
     2281 cggagcagca gcacgctggc actcaagaaa ggcgctgaaa gcttcattga gcttgggcat
     2341 cttgctgggc atgttcttcg taacatggct cccatttttt gtggccaata ttgtccaggc
     2401 ggtgtgcgac tgcatctcac caggcctgtt cgatgtgctc acttggctgg ggtattgcaa
     2461 ttccactatg aaccccatca tatatcccct gttcatgaga gatttcaagc gagcactcgg
     2521 aaggtttctg ccatgcccac ggtgcccacg cgaaaggcaa gcaagtctgg cctccccctc
     2581 actccggacc tcccattcag gacccagacc aggactttca ctccagcaag ttctgcccct
     2641 gccacttcct ccagattcag actctgactc cgatgcgggt tcaggtggct ctagcggact
     2701 ccggcttacg gcccagcttt tgcttcccgg ggaagcaact caggacccac cattgcctac
     2761 tcgggctgcc gccgccgtca atttcttcaa cattgatcct gcagagccag aactcagacc
     2821 gcatcctctg ggtataccaa ccattaggct agcgcccctc tccctccccc ccccctaaac
     2881 ccagctttct tgtacaaagt gggaattccg ataatcaacc tctggattac aaaatttgtg
     2941 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3001 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     3061 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     3121 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     3181 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     3241 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     3301 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
     3361 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     3421 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     3481 ccctttgggc cgcctccccg catcgggaat tcctagagct cgctgatcag cctcgactgt
     3541 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     3601 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     3661 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     3721 agagaatagc aggcatgctg gggagggccg caggaacccc tagtgatgga gttggccact
     3781 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     3841 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     3901 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     3961 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4021 tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc ccttcctttc
     4081 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4141 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4201 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4261 atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc tattcttttg
     4321 atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg atttaacaaa
     4381 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4441 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4501 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4561 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     4621 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4681 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     4741 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     4801 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     4861 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     4921 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     4981 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5041 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5101 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5161 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5221 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5281 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5341 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5401 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5461 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5521 gtggaagccg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5581 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5641 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     5701 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5761 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5821 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5881 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5941 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6001 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6061 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6121 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6181 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6241 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6301 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6361 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6421 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6481 ctcacatgt
//
