LOCUS       sequence-387512-        6210 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses cilia-targeted RhoA inhibitory peptide, tet-inducible.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6210)
  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 6210)
  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-TRE-HTR6-SNAPf-TRIP
FEATURES             Location/Qualifiers
     source          1..6210
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     173..192
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(411..515)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(541..996)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(678..699)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     890..909
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1068..1197
                     /label=AAV2 ITR
     promoter        1272..1586
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1280..1298
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1316..1334
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1351..1369
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1387..1405
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1423..1441
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1458..1476
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1494..1512
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1559..1583
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1618..1656
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     regulatory      3007..3016
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3013..3558
                     /codon_start=1
                     /product="human O6-alkylguanine-DNA-alkyltransferase 
                     variant that forms covalent bonds with benzylguanine 
                     derivatives"
                     /label=SNAP-tag(R)
                     /note="fast-reacting variant, optimized for mammalian 
                     expression"
                     /translation="MDKDCEMKRTTLDSPLGKLELSGCEQGLHRIIFLGKGTSAADAVE
                     VPAPAAVLGGPEPLMQATAWLNAYFHQPEAIEEFPVPALHHPVFQQESFTRQVLWKLLK
                     VVKFGEVISYSHLAALAGNPAATAAVKTALSGNPVPILIPCHRVVQGDLDVGGYEGGLA
                     VKEWLLAHEGHRLGKPGLG"
     misc_feature    3736..4324
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3789..3809)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4207..4218)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4327..4343)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4328..4344)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    4373..4494
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4384..4403)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4438..4457
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(5001..5589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5081..5100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5760..6210,1..341,342..410))
                     /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"
ORIGIN
        1 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
       61 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
      121 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
      181 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
      241 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
      301 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
      361 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
      421 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
      481 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
      541 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
      601 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
      661 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
      721 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
      781 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
      841 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
      901 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
      961 cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc aggctgcgca
     1021 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg cgcgctcgct
     1081 cgctcactga ggccgcccgg gcaaagcccg ggcgtcgggc gacctttggt cgcccggcct
     1141 cagtgagcga gcgagcgcgc agagagggag tggccaactc catcactagg ggttccttgt
     1201 agttaatgat taacccgcca tgctacttat ctacgtagcc atgctctagg aagatcggcc
     1261 gcacgcgtct cgagtttact ccctatcagt gatagagaac gtatgtcgag tttactccct
     1321 atcagtgata gagaacgatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga
     1381 gtttactccc tatcagtgat agagaacgta tgtcgagttt actccctatc agtgatagag
     1441 aacgtatgtc gagtttatcc ctatcagtga tagagaacgt atgtcgagtt tactccctat
     1501 cagtgataga gaacgtatgt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag
     1561 ctcgtttagt gaaccgtcag atcgcctgga gaattcggta ccacgcgtgc caccatgtca
     1621 cgacttgagg aggaactcag aaggagactc actgaaccag tcccagagcc gggcccaacc
     1681 gccaatagca ccccggcctg gggggcaggg ccgccgtcgg ccccgggggg cagcggctgg
     1741 gtggcggccg cgctgtgcgt ggtcatcgcg ctgacggcgg cggccaactc gctgctgatc
     1801 gcgctcatct gcactcagcc cgcgctgcgc aacacgtcca acttcttcct ggtgtcgctc
     1861 ttcacgtctg acctgatggt ggggctggtg gtgatgccgc cggccatgct gaacgcgctg
     1921 tacgggcgct gggtgctggc gcgcggcctc tgcctgctct ggaccgcctt cgacgtgatg
     1981 tgctgcagcg cctccatcct caacctctgc ctcatcagcc tggaccgcta cctgctcatc
     2041 ctctcgccgc tgcgctacaa gctgcgcatg acgcccctgc gtgccctggc cctagtcctg
     2101 ggcgcctgga gcctcgccgc tctcgcctcc ttcctgcccc tgctgctggg ctggcacgag
     2161 ctgggccacg cacggccacc cgtccctggc cagtgccgcc tgctggccag cctgcctttt
     2221 gtccttgtgg cgtcgggcct caccttcttc ctgccctcgg gtgccatatg cttcacctac
     2281 tgcaggatcc tgctagctgc ccgcaagcag gccgtgcagg tggcctccct caccaccggc
     2341 atggccagtc aggcctcgga gacgctgcag gtgcccagga ccccacgccc aggggtggag
     2401 tctgctgaca gcaggcgtct agccacgaag cacagcagga aggccctgaa ggccagcctg
     2461 acgctgggca tcctgctggg catgttcttt gtgacctggt tgcccttctt tgtggccaac
     2521 atagtccagg ccgtgtgcga ctgcatctcc ccaggcctct tcgatgtcct cacatggctg
     2581 ggttactgta acagcaccat gaaccccatc atctacccac tcttcatgcg ggacttcaag
     2641 cgggcgctgg gcaggttcct gccatgtcca cgctgtcccc gggagcgcca ggccagcctg
     2701 gcctcgccat cactgcgcac ctctcacagc ggcccccggc ccggccttag cctacagcag
     2761 gtgctgccgc tgcccctgcc gccggactca gattcggact cagacgcagg ctcaggcggc
     2821 tcctcgggcc tgcggctcac ggcccagctg ctgcttcctg gcgaggccac ccaggacccc
     2881 ccgctgccca ccagggccgc tgccgccgtc aatttcttca acatcgaccc cgcggagccc
     2941 gagctgcggc cgcatccact tggcatcccc acgaacaacg gtacagcggg cccgggatcc
     3001 gcgatcgcca ccatggacaa agactgcgaa atgaagcgca ccaccctgga tagccctctg
     3061 ggcaagctgg aactgtctgg gtgcgaacag ggcctgcacc gtatcatctt cctgggcaaa
     3121 ggaacatctg ccgccgacgc cgtggaagtg cctgccccag ccgccgtgct gggcggacca
     3181 gagccactga tgcaggccac cgcctggctc aacgcctact ttcaccagcc tgaggccatc
     3241 gaggagttcc ctgtgccagc cctgcaccac ccagtgttcc agcaggagag ctttacccgc
     3301 caggtgctgt ggaaactgct gaaagtggtg aagttcggag aggtcatcag ctacagccac
     3361 ctggccgccc tggccggcaa tcccgccgcc accgccgccg tgaaaaccgc cctgagcgga
     3421 aatcccgtgc ccattctgat cccctgccac cgggtggtgc agggcgacct ggacgtgggg
     3481 ggctacgagg gcgggctcgc cgtgaaagag tggctgctgg cccacgaggg ccacagactg
     3541 ggcaagcctg ggctgggtgg aggaggagga tctggaggag gaggatctgc cagcgaggga
     3601 gctgacggag ctatctgtgg ctacaacctg gccaccctgg tcatgctggg accaagcgag
     3661 agggtgttct gcccactgtg cggaccttgc agctccgata tctacgagct gatgtgaact
     3721 agtaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3781 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3841 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3901 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3961 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     4021 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4081 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     4141 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     4201 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     4261 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     4321 ccgcatcgat accgtcgacc tcgacccggg cggccgcttc gagcagacat gataagatac
     4381 attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa
     4441 atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac
     4501 aacaattgca ttcattttat gtttcaggtt cagggggaga tgtgggaggt tttttaaagc
     4561 aagtaaaacc tctacaaatg tggtaaaatc gataaggatc ttcctagagc atggctacgt
     4621 agataagtag catggcgggt taatcattaa ctacaaggaa cccctagtga tggagttggc
     4681 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgrcs
     4741 cccgggcttt gcccgggcgg cctcagtgag taatgaatcg gccaacgcgc ggggagaggc
     4801 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     4861 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     4921 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     4981 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5041 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5101 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5161 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5221 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5281 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5341 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5401 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5461 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5521 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5581 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5641 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5701 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     5761 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     5821 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     5881 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     5941 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6001 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6061 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6121 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6181 gttagctcct tcggtcctcc gatcgttgtc
//
