LOCUS       sequence-403569-        7687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transduce excitatory neurons with AAV to express TRPV1 along with an
            mCherry fluorescence reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7687)
  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 7687)
  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-CamKII-TRPV1-P2A-mCherry
FEATURES             Location/Qualifiers
     source          1..7687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   45..174
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        192..1480
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     regulatory      1490..1499
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4028..4084
                     /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             4085..4792
                     /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(4227..4245)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4495..4514
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     4846..4865
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4994..5134
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4994..5123
                     /label=AAV2 ITR
     rep_origin      5209..5664
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5296..5315)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5506..5527
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5681..5700)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5800..5822
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5860..5878)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5946..6050
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6051..6119,6120..6911)
                     /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(6269..6288)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7082..7670
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7571..7590
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgtcctgca ggcagctgcg
       61 cgctcgctcg ctcactgagg ccgcccgggc gtcgggcgac ctttggtcgc ccggcctcag
      121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc
      181 gcacgcgtta acattatggc cttaggtcac ttcatctcca tggggttctt cttctgattt
      241 tctagaaaat gagatggggg tgcagagagc ttcctcagtg acctgcccag ggtcacatca
      301 gaaatgtcag agctagaact tgaactcaga ttactaatct taaattccat gccttggggg
      361 catgcaagta cgatatacag aaggagtgaa ctcattaggg cagatgacca atgagtttag
      421 gaaagaagag tccagggcag ggtacatcta caccacccgc ccagccctgg gtgagtccag
      481 ccacgttcac ctcattatag ttgcctctct ccagtcctac cttgacggga agcacaagca
      541 gaaactggga caggagcccc aggagaccaa atcttcatgg tccctctggg aggatgggtg
      601 gggagagctg tggcagaggc ctcaggaggg gccctgctgc tcagtggtga cagatagggg
      661 tgagaaagca gacagagtca ttccgtcagc attctgggtc tgtttggtac ttcttctcac
      721 gctaaggtgg cggtgtgata tgcacaatgg ctaaaaagca gggagagctg gaaagaaaca
      781 aggacagaga cagaggccaa gtcaaccaga ccaattccca gaggaagcaa agaaaccatt
      841 acagagacta caagggggaa gggaaggaga gatgaattag cttcccctgt aaaccttaga
      901 acccagctgt tgccagggca acggggcaat acctgtctct tcagaggaga tgaagttgcc
      961 agggtaacta catcctgtct ttctcaagga ccatcccaga atgtggcacc cactagccgt
     1021 taccatagca actgcctctt tgccccactt aatcccatcc cgtctgttaa aagggcccta
     1081 tagttggagg tgggggaggt aggaagagcg atgatcactt gtggactaag tttgttcgca
     1141 tccccttctc caaccccctc agtacatcac cctgggggaa cagggtccac ttgctcctgg
     1201 gcccacacag tcctgcagta ttgtgtatat aaggccaggg caaagaggag caggttttaa
     1261 agtgaaaggc aggcaggtgt tggggaggca gttaccgggg caacgggaac agggcgtttc
     1321 ggaggtggtt gccatgggga cctggatgct gacgaaggct cgcgaggctg tgagcagcca
     1381 cagtgccctg ctcagaagcc ccaagctcgt cagtcaagcc ggttctccgt ttgcactcag
     1441 gagcacgggc aggcgagtgg cccctagttc tgggggcagc gggggatccg ccaccatgga
     1501 acaacgggct agcttagact cagaggagtc tgagtcccca ccccaagaga actcctgcct
     1561 ggaccctcca gacagagacc ctaactgcaa gccacctcca gtcaagcccc acatcttcac
     1621 taccaggagt cgtacccggc tttttgggaa gggtgactcg gaggaggcct ctcccctgga
     1681 ctgcccttat gaggaaggcg ggctggcttc ctgccctatc atcactgtca gctctgttct
     1741 aactatccag aggcctgggg atggacctgc cagtgtcagg ccgtcatccc aggactccgt
     1801 ctccgctggt gagaagcccc cgaggctcta tgatcgcagg agcatcttcg atgctgtggc
     1861 tcagagtaac tgccaggagc tggagagcct gctgcccttc ctgcagagga gcaagaagcg
     1921 cctgactgac agcgagttca aagacccaga gacaggaaag acctgtctgc taaaagccat
     1981 gctcaatctg cacaatgggc agaatgacac catcgctctg ctcctggacg ttgcccggaa
     2041 gacagacagc ctgaagcagt ttgtcaatgc cagctacaca gacagctact acaagggcca
     2101 gacagcactg cacattgcca ttgaacggcg gaacatgacg ctggtgaccc tcttggtgga
     2161 gaatggagca gatgtccagg ctgcggctaa cggggacttc ttcaagaaaa ccaaagggag
     2221 gcctggcttc tactttggtg agctgcccct gtccctggct gcgtgcacca accagctggc
     2281 cattgtgaag ttcctgctgc agaactcctg gcagcctgca gacatcagcg cccgggactc
     2341 agtgggcaac acggtgcttc atgccctggt ggaggtggca gataacacag ttgacaacac
     2401 caagttcgtg acaagcatgt acaacgagat cttgatcctg ggggccaaac tccaccccac
     2461 gctgaagctg gaagagatca ccaacaggaa ggggctcacg ccactggctc tggctgctag
     2521 cagtgggaag atcggggtct tggcctacat tctccagagg gagatccatg aacccgagtg
     2581 ccgacaccta tccaggaagt tcaccgaatg ggcctatggg ccagtgcact cctcccttta
     2641 tgacctgtcc tgcattgaca cctgtgaaaa gaactcggtt ctggaggtga tcgcttacag
     2701 cagcagtgag acccctaacc gtcatgacat gcttctcgtg gaacccttga accgactcct
     2761 acaggacaag tgggacagat ttgtcaagcg catcttctac ttcaacttct tcgtctactg
     2821 cttgtatatg atcatcttca ccgcggctgc ctactatcgg cctgtggaag gcttgccccc
     2881 ctataagctg aaaaacaccg ttggggacta tttccgagtc accggagaga tcttgtctgt
     2941 gtcaggagga gtctacttct tcttccgagg gattcaatat ttcctgcaga ggcgaccatc
     3001 cctcaagagt ttgtttgtgg acagctacag tgagatactt ttctttgtac agtcgctgtt
     3061 catgctggtg tctgtggtac tgtacttcag ccaacgcaag gagtatgtgg cttccatggt
     3121 gttctccctg gccatgggct ggaccaacat gctctactat acccgaggat tccagcagat
     3181 gggcatctat gctgtcatga ttgagaagat gatcctcaga gacctgtgcc ggtttatgtt
     3241 cgtctacctc gtgttcttgt ttggattttc cacagctgtg gtgacactga ttgaggatgg
     3301 gaagaataac tctctgccta tggagtccac accacacaag tgccgggggt ctgcctgcaa
     3361 gccaggtaac tcttacaaca gcctgtattc cacatgtctg gagctgttca agttcaccat
     3421 cggcatgggc gacctggagt tcactgagaa ctacgacttc aaggctgtct tcatcatcct
     3481 gttactggcc tatgtgattc tcacctacat ccttctgctc aacatgctca ttgctctcat
     3541 gggtgagacc gtcaacaaga ttgcacaaga gagcaagaac atctggaagc tgcagagagc
     3601 catcaccatc ctggatacag agaagagctt cctgaagtgc atgaggaagg ccttccgctc
     3661 tggcaagctg ctgcaggtgg ggttcactcc tgacggcaag gatgactacc ggtggtgttt
     3721 cagggtggac gaggtaaact ggactacctg gaacaccaat gtgggtatca tcaacgagga
     3781 cccaggcaac tgtgagggcg tcaagcgcac cctgagcttc tccctgaggt caggccgagt
     3841 ttcagggaga aactggaaga actttgccct ggttcccctt ctgagggatg caagcactcg
     3901 agatagacat gccacccagc aggaagaagt tcaactgaag cattatacgg gctcccttaa
     3961 gccagaggat gctgaggttt tcaaggattc catggtccca ggggagaaag gcgcgcccgg
     4021 ctccggagcc acgaacttct ctctgttaaa gcaagcagga gacgtggaag aaaaccccgg
     4081 tcccgtgagc aagggcgagg aggataacat ggccatcatc aaggagttca tgcgcttcaa
     4141 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     4201 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     4261 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     4321 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     4381 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     4441 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     4501 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     4561 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     4621 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     4681 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     4741 acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt aaggatccac
     4801 cggatctaga taaggcctca cctgcgatct cgatgcttta tttgtgaaat ttgtgatgct
     4861 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     4921 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaaactag tcacgtgcgg
     4981 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     5041 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     5101 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     5161 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     5221 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     5281 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     5341 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     5401 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     5461 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     5521 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     5581 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     5641 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     5701 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     5761 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     5821 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     5881 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5941 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6001 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     6061 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     6121 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6181 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6241 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     6301 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6361 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6421 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6481 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6541 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     6601 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6661 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6721 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6781 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6841 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6901 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6961 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7021 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7081 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7141 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7201 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7261 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7321 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7381 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7441 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7501 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7561 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7621 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7681 acgcggc
//
