LOCUS       sequence-398423-        8110 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In the presence of Cre, drives neuron-specific expression of TRPV1 
            with fluorescence reporter mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8110)
  AUTHORS   Yang Y, Pacia CP, Ye D, Zhu L, Baek H, Yue Y, Yuan J, Miller MJ, Cui
            J, Culver JP, Bruchas MR, Chen H
  TITLE     Sonothermogenetics for noninvasive and cell-type specific deep brain
            neuromodulation.
  JOURNAL   Brain Stimul. 2021 Jul-Aug;14(4):790-800. doi: 
            10.1016/j.brs.2021.04.021. Epub 2021 May 11.
  PUBMED    33989819
REFERENCE   2  (bases 1 to 8110)
  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"; journalName: "Brain 
            Stimul. 2021 Jul-Aug"; volume: "14(4)"; pages: "790-800. doi: 
            10.1016/j.brs.2021.04.021. Epub 2021 May 11"
COMMENT     Sequence Label: AAV-Syn-DIO-TRPV1-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..8110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   20..149
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        194..641
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(679..712)
                     /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(763..796)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(805..1512)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     primer_bind     complement(1083..1102)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1352..1370
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(1513..1569)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     regulatory      4098..4107
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    4114..4147
                     /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    4198..4231
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    4256..4844
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4309..4329)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4727..4738)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4876..5352
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(5075..5094)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5392..5532
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5392..5521
                     /label=AAV2 ITR
     rep_origin      5607..6062
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5694..5713)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5904..5925
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(6079..6098)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6198..6220
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6258..6276)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6344..6448
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6449..6517,6518..7309)
                     /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(6667..6686)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7480..8068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7969..7988
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tggccttttg ctcacatgtc ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc
       61 cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg
      121 agtggccaac tccatcacta ggggttcctg cggccgcacg cgtgtgtcta gactgcagag
      181 ggccctgcgt atgagtgcaa gtgggtttta ggaccaggat gaggcggggt gggggtgcct
      241 acctgacgac cgaccccgac ccactggaca agcacccaac ccccattccc caaattgcgc
      301 atcccctatc agagaggggg aggggaaaca ggatgcggcg aggcgcgtgc gcactgccag
      361 cttcagcacc gcggacagtg ccttcgcccc cgcctggcgg cgcgcgccac cgccgcctca
      421 gcactgaagg cgcgctgacg tcactcgccg gtcccccgca aactcccctt cccggccacc
      481 ttggtcgcgt ccgcgccgcc gccggcccag ccggaccgca ccacgcgagg cgcgagatag
      541 gggggcacgg gcgcgaccat ctgcgctgcg gcgccggcga ctcagcgctg cctcagtctg
      601 cggtgggcag cggaggagtc gtgtcgtgcc tgagagcgca gtcgagaagg taccggatcc
      661 tctagagtcg actccggaat aacttcgtat aggatacttt atacgaagtt atgcagaatg
      721 gtagctggat tgtagctgct attagcaata tgaaacctct taataacttc gtatagcata
      781 cattatacga agttatggcg cgccttactt gtacagctcg tccatgccgc cggtggagtg
      841 gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
      901 gtccaacttg atgttgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta
      961 ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat
     1021 ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat
     1081 ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt
     1141 gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc
     1201 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcaagtagtc
     1261 ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag
     1321 gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt
     1381 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga
     1441 gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc
     1501 gcccttgctc acgggaccgg ggttttcttc cacgtctcct gcttgcttta acagagagaa
     1561 gttcgtggct ccggagccgg gcgcgccttt ctcccctggg accatggaat ccttgaaaac
     1621 ctcagcatcc tctggcttaa gggagcccgt ataatgcttc agttgaactt cttcctgctg
     1681 ggtggcatgt ctatctcgag tgcttgcatc cctcagaagg ggaaccaggg caaagttctt
     1741 ccagtttctc cctgaaactc ggcctgacct cagggagaag ctcagggtgc gcttgacgcc
     1801 ctcacagttg cctgggtcct cgttgatgat acccacattg gtgttccagg tagtccagtt
     1861 tacctcgtcc accctgaaac accaccggta gtcatccttg ccgtcaggag tgaaccccac
     1921 ctgcagcagc ttgccagagc ggaaggcctt cctcatgcac ttcaggaagc tcttctctgt
     1981 atccaggatg gtgatggctc tctgcagctt ccagatgttc ttgctctctt gtgcaatctt
     2041 gttgacggtc tcacccatga gagcaatgag catgttgagc agaaggatgt aggtgagaat
     2101 cacataggcc agtaacagga tgatgaagac agccttgaag tcgtagttct cagtgaactc
     2161 caggtcgccc atgccgatgg tgaacttgaa cagctccaga catgtggaat acaggctgtt
     2221 gtaagagtta cctggcttgc aggcagaccc ccggcacttg tgtggtgtgg actccatagg
     2281 cagagagtta ttcttcccat cctcaatcag tgtcaccaca gctgtggaaa atccaaacaa
     2341 gaacacgagg tagacgaaca taaaccggca caggtctctg aggatcatct tctcaatcat
     2401 gacagcatag atgcccatct gctggaatcc tcgggtatag tagagcatgt tggtccagcc
     2461 catggccagg gagaacacca tggaagccac atactccttg cgttggctga agtacagtac
     2521 cacagacacc agcatgaaca gcgactgtac aaagaaaagt atctcactgt agctgtccac
     2581 aaacaaactc ttgagggatg gtcgcctctg caggaaatat tgaatccctc ggaagaagaa
     2641 gtagactcct cctgacacag acaagatctc tccggtgact cggaaatagt ccccaacggt
     2701 gtttttcagc ttataggggg gcaagccttc cacaggccga tagtaggcag ccgcggtgaa
     2761 gatgatcata tacaagcagt agacgaagaa gttgaagtag aagatgcgct tgacaaatct
     2821 gtcccacttg tcctgtagga gtcggttcaa gggttccacg agaagcatgt catgacggtt
     2881 aggggtctca ctgctgctgt aagcgatcac ctccagaacc gagttctttt cacaggtgtc
     2941 aatgcaggac aggtcataaa gggaggagtg cactggccca taggcccatt cggtgaactt
     3001 cctggatagg tgtcggcact cgggttcatg gatctccctc tggagaatgt aggccaagac
     3061 cccgatcttc ccactgctag cagccagagc cagtggcgtg agccccttcc tgttggtgat
     3121 ctcttccagc ttcagcgtgg ggtggagttt ggcccccagg atcaagatct cgttgtacat
     3181 gcttgtcacg aacttggtgt tgtcaactgt gttatctgcc acctccacca gggcatgaag
     3241 caccgtgttg cccactgagt cccgggcgct gatgtctgca ggctgccagg agttctgcag
     3301 caggaacttc acaatggcca gctggttggt gcacgcagcc agggacaggg gcagctcacc
     3361 aaagtagaag ccaggcctcc ctttggtttt cttgaagaag tccccgttag ccgcagcctg
     3421 gacatctgct ccattctcca ccaagagggt caccagcgtc atgttccgcc gttcaatggc
     3481 aatgtgcagt gctgtctggc ccttgtagta gctgtctgtg tagctggcat tgacaaactg
     3541 cttcaggctg tctgtcttcc gggcaacgtc caggagcaga gcgatggtgt cattctgccc
     3601 attgtgcaga ttgagcatgg cttttagcag acaggtcttt cctgtctctg ggtctttgaa
     3661 ctcgctgtca gtcaggcgct tcttgctcct ctgcaggaag ggcagcaggc tctccagctc
     3721 ctggcagtta ctctgagcca cagcatcgaa gatgctcctg cgatcataga gcctcggggg
     3781 cttctcacca gcggagacgg agtcctggga tgacggcctg acactggcag gtccatcccc
     3841 aggcctctgg atagttagaa cagagctgac agtgatgata gggcaggaag ccagcccgcc
     3901 ttcctcataa gggcagtcca ggggagaggc ctcctccgag tcacccttcc caaaaagccg
     3961 ggtacgactc ctggtagtga agatgtgggg cttgactgga ggtggcttgc agttagggtc
     4021 tctgtctgga gggtccaggc aggagttctc ttggggtggg gactcagact cctctgagtc
     4081 taagctagcc cgttgttcca tggtggcgct agcataactt cgtataaagt atcctatacg
     4141 aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt gcaaagaata
     4201 acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta tcgataatca
     4261 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     4321 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     4381 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc
     4441 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg
     4501 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc
     4561 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg
     4621 cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc tgctcgccta
     4681 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc
     4741 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct
     4801 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat accgagcgct
     4861 gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct cctggccctg
     4921 gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg catcattttg
     4981 tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg gagcaagggg
     5041 caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag ctggagtgca
     5101 gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct
     5161 cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta atttttgttt
     5221 ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc ctaatctcag
     5281 gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac cactgctccc
     5341 ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg caggaacccc
     5401 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     5461 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     5521 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     5581 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     5641 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     5701 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     5761 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     5821 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     5881 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     5941 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     6001 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     6061 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     6121 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     6181 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     6241 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     6301 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     6361 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     6421 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     6481 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     6541 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     6601 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     6661 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     6721 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     6781 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     6841 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     6901 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     6961 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     7021 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     7081 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     7141 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     7201 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     7261 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     7321 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     7381 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     7441 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     7501 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     7561 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     7621 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     7681 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     7741 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     7801 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     7861 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     7921 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     7981 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     8041 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     8101 ggccttttgc
//
