LOCUS       sequence-271479-        3219 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV vector for Cre-dependent expression of Venus-tagged parapinopsin
            driven by the CAG promoter. Used for photoswitchable control of 
            inhibitory GPCR signaling cascades..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3219)
  AUTHORS   Copits BA, Gowrishankar R, O'Neill PR, Li JN, Girven KS, Yoo JJ, 
            Meshik X, Parker KE, Spangler SM, Elerding AJ, Brown BJ, Shirley SE,
            Ma KKL, Vasquez AM, Stander MC, Kalyanaraman V, Vogt SK, Samineni 
            VK, Patriarchi T, Tian L, Gautam N, Sunahara RK, Gereau RW 4th, 
            Bruchas MR
  TITLE     A photoswitchable GPCR-based opsin for presynaptic inhibition.
  JOURNAL   Neuron. 2021 Jun 2;109(11):1791-1809.e11. doi: 
            10.1016/j.neuron.2021.04.026. Epub 2021 May 11.
  PUBMED    33979635
REFERENCE   2  (bases 1 to 3219)
  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.2021.04.026"; journalName: "Neuron"; date: 
            "2021-06-2- 2"; volume: "109"; issue: "11"; pages: "1791-1809.e11"
COMMENT     Sequence Label: pAAV-CAG-DIO-PPO-Venus
FEATURES             Location/Qualifiers
     source          1..3219
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     polyA_signal    31..142
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     131..148
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     misc_feature    175..763
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             281..292
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     690..710
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     protein_bind    complement(788..821)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(872..905)
                     /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)."
     regulatory      1957..1966
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1963..1965,1966..1968,1969..2682)
                     /codon_start=1
                     /product="Venus YFP with monomerizing A206K mutation (Nagai
                     et al., 2002; Kremers et al., 2006)"
                     /label=mVenus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2008..2029)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2269..2288)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2616..2637
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    2716..2749
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    2800..2833
                     /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)."
     primer_bind     complement(2884..2903)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     primer_bind     complement(2965..2987)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 cagaatagaa tgacactact cagacaatgc gatgcaattt cntcatttta ttaggaaagg
       61 acagtgggag tggcactttn cagggtcaag gaaggcacgg gggaggggca aacaacagat
      121 ggctggcaac tagaaggcac agtcgaggct gatcagcgag tcgacggtat cgatgcgggg
      181 aggcggccca aagggagatc cgactcgtct gagggcgaag gcgaagacgc ggaagaggcc
      241 gcagagccgg cagcaggccg cgggaaggaa ggtccgctgg attgagggcc gaagggacgt
      301 agcagaagga cgtcccgcgc agaatccagg tggcaacaca ggcgagcagc caaggaaagg
      361 acgatgattt ccccgacaac accacggaat tgtcagtgcc caacagccga gcccctgtcc
      421 agcagcgggc aaggcaggcg gcgatgagtt ccgccgtggc aatagggagg gggaaagcga
      481 aagtcccgga aaggagctga caggtggtgg caatgcccca accagtgggg gttgcgtcag
      541 caaacacagt gcacaccacg ccacgttgcc tgacaacggg ccacaactcc tcataaagag
      601 acagcaacca ggatttatac aaggaggaga aaatgaaagc catacgggaa gcaatagcat
      661 gatacaaagg cattaaagca gcgtatccac atagcgtaaa aggagcaaca tagttaagaa
      721 taccagtcaa tctttcacaa attttgtaat ccagaggttg attatcgata agcttgatat
      781 cgaattcata acttcgtata gcatacatta tacgaagtta ttctttgcac cattctaaag
      841 aataacagtg ataatttctg ggttaaggca aataacttcg tataggatac tttatacgaa
      901 gttatgctag caccggtgga attcatggag aacttgacct cgctcgacct cctgcccaac
      961 ggcgaggtcc cgttgatgcc ccgctacggc ttcaccatcc tcgccgtgat catggccgtg
     1021 ttcaccatcg cctcgctcgt gctcaacagc accgtcgtca tcgtcaccct gcgccaccgc
     1081 cagctacgcc acccgctcaa cttctcgctc gtcaacctcg ccgtggcgga cctgggcgtc
     1141 acggtgttcg gcgccagcct cgtcgtggag accaacgccg tcgggtactt caacctcggc
     1201 cgcgtcggct gcgtcatcga agggttcgcc gtcgctttct tcggcatcgc cgctctgtgc
     1261 acgatcgccg tgatcgccgt cgatcgcttc gtggtggtgt gcaagccgct gggcacgctg
     1321 atgttcacgc ggcgccacgc gctgctgggc atcgcctggg cctggctctg gtcgttcgtg
     1381 tggaacacgc cgccgctctt cggctggggc agctacgagc tggagggcgt gcggacgtcg
     1441 tgcgcgcccg actggtacag ccgcgacccc gccaacgtgt cgtacattac gagctacttc
     1501 gccttctgct tcgccatccc cttcctcgtc atcgtggtgg cgtacggccg cctcatgtgg
     1561 accctccacc aggtggccaa gctggggatg ggcgagagcg gcagcaccgc caaggcggag
     1621 gcgcaggtgt cgcgcatggt ggtggtcatg gtggtggcct tcctcgtctg ctggctgccc
     1681 tacgcgctct tcgccatgat cgtggtgacc aagcccgacg tgtacatcga cccggtcatc
     1741 gccacactgc ccatgtacct gaccaagacg agcacggtct acaaccccat catctacatc
     1801 ttcatgaacc gccagttccg ggactgcgcc gtgcccttcc tgctctgcgg acgcaacccc
     1861 tgggcagagc cttcctcgga gtcggcgacg gcggcctcca cgtcggccac cagcgtgacc
     1921 ctggcgtccg cgccggggca ggtctccccg agcgcggccg ccatggtgag caagggcgag
     1981 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     2041 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     2101 ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgggc
     2161 tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
     2221 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     2281 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     2341 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     2401 aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc
     2461 aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta ccagcagaac
     2521 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc
     2581 aaactgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     2641 gccgccggga tcactctcgg catggacgag ctgtacaagt aatctagagg gcccgtttaa
     2701 acccgctggc gcgccataac ttcgtataat gtatgctata cgaagttatt aagaggtttc
     2761 atattgctaa tagcagctac aatccagcta ccattctgca taacttcgta taaagtatcc
     2821 tatacgaagt tattccggag tcgactctag aggatccaat tctttgccaa aatgatgaga
     2881 cagcacaata accagcacgt tgcccaggag ctgtaggaaa aagaagaagg catgaacatg
     2941 gttagcagag gctctagagc cgccggtcac acgccagaag ccgaaccccg ccctgccccg
     3001 tcccccccga aggcagccgt ccccccgcgg acagccccga gccgctcccc ccgcacaaag
     3061 ggccctcccg gagcccctca aggctttcac gcagccacag aaaagaaaca agccgtcatt
     3121 aaaccaagcg ctaattacag cccggaggag aagggccgtc ccgcccgctc acctgtggga
     3181 gtaacgcggt cagtcagagc cggggcgggc ggcgcgagg
//
