LOCUS       sequence-209246-        5798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of ChETA (hChR2 with E123T/H134R mutations) fused to 
            HA tag driven by hSynapsin promoter in Cre-ON manner for optogenetic
            activation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5798)
  AUTHORS   Li SJ, Vaughan A, Sturgill JF, Kepecs A
  TITLE     A Viral Receptor Complementation Strategy to Overcome CAV-2 Tropism 
            for Efficient Retrograde Targeting of Neurons.
  JOURNAL   Neuron. 2018 Jun 6;98(5):905-917.e5. doi: 
            10.1016/j.neuron.2018.05.028.
  PUBMED    29879392
REFERENCE   2  (bases 1 to 5798)
  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.neuron.2018.05"; journalName: "Neuron"; date: "2018-06-6-
            6"; volume: "98"; issue: "5"; pages: "905-917.e5"
COMMENT     Sequence Label: pAAV-hSyn-DIO {ChETA}on-WPRE
FEATURES             Location/Qualifiers
     source          1..5798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..142
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   13..142
                     /label=AAV2 ITR
     promoter        187..634
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(672..705)
                     /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(756..789)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(802..828)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             complement(802..828)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             complement(847..1773)
                     /codon_start=1
                     /product="H134R mutant of an N-terminal fragment of 
                     channelrhodopsin-2 from the alga Chlamydomonas reinhardtii 
                     (Nagel et al., 2005)"
                     /label=ChR2(H134R)
                     /note="blue light-activated cation channel"
                     /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT
                     NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN
                     PSMLYLATGHRVQWLRYATWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG
                     ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS
                     WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI
                     RKTTKLNIGGTEIEVETLVEDEAEAGAV"
     regulatory      1770..1779
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1784..1817
                     /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    1868..1901
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1926..2514
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1979..1999)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2397..2408)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2546..3022
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2745..2764)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3062..3202
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3062..3191
                     /label=AAV2 ITR
     rep_origin      3277..3732
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3364..3383)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3574..3595
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3749..3768)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3868..3890
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3928..3946)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4014..4118
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4119..4187,4188..4979)
                     /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(4337..4356)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5150..5738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5639..5658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca gagggccctg
      181 cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac
      241 gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct
      301 atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc
      361 accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga
      421 aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg
      481 cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca
      541 cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg
      601 cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga aggtaccgga tcctctagag
      661 tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg
      721 gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc atacattata
      781 cgaagttatg gcgcgtaatt aggcgtagtc gggcacgtcg taggggtaca tggtggcggc
      841 cgctggcacg gctccggcct cggcttcgtc ttcgacgaga gtctcgacct cgatctccgt
      901 tccgccgatg ttcagtttgg tggttttgcg gatatctccg tgaatcaata tgtgctcgtg
      961 gatcaggacg cgcaggtagt gtcccaacaa cccccaacaa tttttactca tcagatcaat
     1021 aatcgtgtga cctacggtgg agccatagac gctcaggacg ccaaaacctt cgggccccaa
     1081 aatgaagaga attgggaaca taccccagct cacgaaaaac agccatgcca tgccggtcac
     1141 gacctggcgg caccgaccct ttggcacagt atgataaccc tcgatatatg ctttggcggc
     1201 gtgaaaaaat gtgttcgcgc catagcacaa tccaagacaa aagaagatga ctttaacata
     1261 gccggttgcc atggcgctgg tagcccccca cacgatagtc ccgatgtctg agacaaggag
     1321 tcccatggtt ctcctgctgt agtcgttgct caggccggtg aggttgctca ggcggataag
     1381 gatgacagga caagtgagca gccaggttgc atagcgcagc cactgcaccc ggtgtcctgt
     1441 ggcaaggtag agcatagagg gattcttaaa ctcaaaaaag aactcgagaa tcaccttaac
     1501 catttcaatg gcgcacacat agatctcctc ccagccgcat gtagatttcc aggtttggta
     1561 ggcatagaac atcagcagca aaatgctgaa tcctgctgca agccactgca ggacatttga
     1621 cgcggtctga gcgccgttcg tgccgcgaga ttcaatccat ccggcacagt aacattgatc
     1681 ctcagggacc aggacggacc cgttcaccac cacaggatta gtaacgaaca aaagttcgcg
     1741 tccgacggca gacaaagcgc cgccatagtc catggtggct agcataactt cgtataaagt
     1801 atcctatacg aagttatttg ccttaaccca gaaattatca ctgttattct ttagaatggt
     1861 gcaaagaata acttcgtata atgtatgcta tacgaagtta tgaattcgat atcaagctta
     1921 tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg
     1981 ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt
     2041 cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg
     2101 agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc
     2161 ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc
     2221 tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc
     2281 ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc tttccttggc
     2341 tgctcgccta tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg
     2401 ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc
     2461 gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcatcgat
     2521 accgagcgct gctcgagaga tctacgggtg gcatccctgt gacccctccc cagtgcctct
     2581 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     2641 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     2701 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     2761 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     2821 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     2881 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     2941 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3001 cactgctccc ttccctgtcc ttctgatttt gtaggtaacc acgtgcggac cgagcggccg
     3061 caggaacccc tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag
     3121 gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag
     3181 cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3241 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3301 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3361 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     3421 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3481 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     3541 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     3601 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     3661 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     3721 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     3781 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3841 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3901 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     3961 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4021 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4081 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4141 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4201 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4261 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4321 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4381 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     4441 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     4501 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     4561 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     4621 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     4681 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     4741 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     4801 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     4861 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     4921 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     4981 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5041 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5101 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5161 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5221 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5281 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5341 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     5401 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     5461 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     5521 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     5581 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     5641 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     5701 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     5761 tacggttcct ggccttttgc tggccttttg ctcacatg
//
