LOCUS       sequence-398422-        7891 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transduce all neuron types with AAV to express TRPV1 along with a 
            DsRed fluorescence reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7891)
  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 7891)
  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-TRPV1-P2A-DsRed
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      654..663
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3192..3248
                     /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"
     CDS             3261..3935
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(3441..3461)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     3853..3876
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     misc_feature    3936..3992
                     /label=MCS
                     /note="multiple cloning site"
     misc_feature    4018..4606
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4071..4091)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4489..4500)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4638..5114
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4837..4856)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5154..5294
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5154..5283
                     /label=AAV2 ITR
     rep_origin      5369..5824
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5456..5475)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5666..5687
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5841..5860)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5960..5982
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6020..6038)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6106..6210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6211..6279,6280..7071)
                     /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(6429..6448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7242..7830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7731..7750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca
      181 agtgggtttt aggaccagga tgaggcgggg kgggggkgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgagaag gtaccggatc ctctagagtc gacgccacca
      661 tggaacaacg ggctagctta gactcagagg agtctgagtc cccaccccaa gagaactcct
      721 gcctggaccc tccagacaga gaccctaact gcaagccacc tccagtcaag ccccacatct
      781 tcactaccag gagtcgtacc cggctttttg ggaagggtga ctcggaggag gcctctcccc
      841 tggactgccc ttatgaggaa ggcgggctgg cttcctgccc tatcatcact gtcagctctg
      901 ttctaactat ccagaggcct ggggatggac ctgccagtgt caggccgtca tcccaggact
      961 ccgtctccgc tggtgagaag cccccgaggc tctatgatcg caggagcatc ttcgatgctg
     1021 tggctcagag taactgccag gagctggaga gcctgctgcc cttcctgcag aggagcaaga
     1081 agcgcctgac tgacagcgag ttcaaagacc cagagacagg aaagacctgt ctgctaaaag
     1141 ccatgctcaa tctgcacaat gggcagaatg acaccatcgc tctgctcctg gacgttgccc
     1201 ggaagacaga cagcctgaag cagtttgtca atgccagcta cacagacagc tactacaagg
     1261 gccagacagc actgcacatt gccattgaac ggcggaacat gacgctggtg accctcttgg
     1321 tggagaatgg agcagatgtc caggctgcgg ctaacgggga cttcttcaag aaaaccaaag
     1381 ggaggcctgg cttctacttt ggtgagctgc ccctgtccct ggctgcgtgc accaaccagc
     1441 tggccattgt gaagttcctg ctgcagaact cctggcagcc tgcagacatc agcgcccggg
     1501 actcagtggg caacacggtg cttcatgccc tggtggaggt ggcagataac acagttgaca
     1561 acaccaagtt cgtgacaagc atgtacaacg agatcttgat cctgggggcc aaactccacc
     1621 ccacgctgaa gctggaagag atcaccaaca ggaaggggct cacgccactg gctctggctg
     1681 ctagcagtgg gaagatcggg gtcttggcct acattctcca gagggagatc catgaacccg
     1741 agtgccgaca cctatccagg aagttcaccg aatgggccta tgggccagtg cactcctccc
     1801 tttatgacct gtcctgcatt gacacctgtg aaaagaactc ggttctggag gtgatcgctt
     1861 acagcagcag tgagacccct aaccgtcatg acatgcttct cgtggaaccc ttgaaccgac
     1921 tcctacagga caagtgggac agatttgtca agcgcatctt ctacttcaac ttcttcgtct
     1981 actgcttgta tatgatcatc ttcaccgcgg ctgcctacta tcggcctgtg gaaggcttgc
     2041 ccccctataa gctgaaaaac accgttgggg actatttccg agtcaccgga gagatcttgt
     2101 ctgtgtcagg aggagtctac ttcttcttcc gagggattca atatttcctg cagaggcgac
     2161 catccctcaa gagtttgttt gtggacagct acagtgagat acttttcttt gtacagtcgc
     2221 tgttcatgct ggtgtctgtg gtactgtact tcagccaacg caaggagtat gtggcttcca
     2281 tggtgttctc cctggccatg ggctggacca acatgctcta ctatacccga ggattccagc
     2341 agatgggcat ctatgctgtc atgattgaga agatgatcct cagagacctg tgccggttta
     2401 tgttcgtcta cctcgtgttc ttgtttggat tttccacagc tgtggtgaca ctgattgagg
     2461 atgggaagaa taactctctg cctatggagt ccacaccaca caagtgccgg gggtctgcct
     2521 gcaagccagg taactcttac aacagcctgt attccacatg tctggagctg ttcaagttca
     2581 ccatcggcat gggcgacctg gagttcactg agaactacga cttcaaggct gtcttcatca
     2641 tcctgttact ggcctatgtg attctcacct acatccttct gctcaacatg ctcattgctc
     2701 tcatgggtga gaccgtcaac aagattgcac aagagagcaa gaacatctgg aagctgcaga
     2761 gagccatcac catcctggat acagagaaga gcttcctgaa gtgcatgagg aaggccttcc
     2821 gctctggcaa gctgctgcag gtggggttca ctcctgacgg caaggatgac taccggtggt
     2881 gtttcagggt ggacgaggta aactggacta cctggaacac caatgtgggt atcatcaacg
     2941 aggacccagg caactgtgag ggcgtcaagc gcaccctgag cttctccctg aggtcaggcc
     3001 gagtttcagg gagaaactgg aagaactttg ccctggttcc ccttctgagg gatgcaagca
     3061 ctcgagatag acatgccacc cagcaggaag aagttcaact gaagcattat acgggctccc
     3121 ttaagccaga ggatgctgag gttttcaagg attccatggt cccaggggag aaaggcgcgc
     3181 ccggctccgg agccacgaac ttctctctgt taaagcaagc aggagacgtg gaagaaaacc
     3241 ccggtcccat ggatcccacc atggcctcct ccgagaacgt catcaccgag ttcatgcgct
     3301 tcaaggtgcg catggagggc accgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     3361 agggccgccc ctacgagggc cacaacaccg tgaagctgaa ggtgaccaag ggcggccccc
     3421 tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag gtgtacgtga
     3481 agcaccccgc cgacatcccc gactacaaga agctgtcctt ccccgagggc ttcaagtggg
     3541 agcgcgtgat gaacttcgag gacggcggcg tggcgaccgt gacccaggac tcctccctgc
     3601 aggacggctg cttcatctac aaggtgaagt tcatcggcgt gaacttcccc tccgacggcc
     3661 ccgtgatgca gaagaagacc atgggctggg aggcctccac cgagcgcctg tacccccgcg
     3721 acggcgtgct gaagggcgag acccacaagg ccctgaagct gaaggacggc ggccactacc
     3781 tggtggagtt caagtccatc tacatggcca agaagcccgt gcagctgccc ggctactact
     3841 acgtggacgc caagctggac atcacctccc acaacgagga ctacaccatc gtggagcagt
     3901 acgagcgcac cgagggccgc caccacctgt tcctgagatc tcgagctcaa gcttcgaatt
     3961 ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc tagataagct tatcgataat
     4021 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     4081 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     4141 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     4201 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     4261 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     4321 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     4381 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     4441 tatgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     4501 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     4561 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg
     4621 ctgctcgaga gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc
     4681 tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt
     4741 tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg
     4801 ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg
     4861 cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc
     4921 ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt
     4981 ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc
     5041 aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc
     5101 ccttccctgt ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc
     5161 cctagtgatg gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg
     5221 accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg
     5281 cagctgcctg caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
     5341 acaccgcata cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg
     5401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct
     5461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5521 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5581 gatttgggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5701 tctatctcgg gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta
     5761 aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca
     5821 attttatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     5881 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     5941 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     6001 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     6061 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     6121 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     6181 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     6241 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     6301 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     6361 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     6421 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     6481 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     6541 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     6601 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     6661 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     6721 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     6781 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     6841 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     6901 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     6961 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     7021 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     7081 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     7141 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     7201 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     7261 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     7321 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     7381 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     7441 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     7501 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     7561 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     7621 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     7681 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     7741 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     7801 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     7861 ctggcctttt gctggccttt tgctcacatg t
//
