LOCUS       sequence-276180-        7388 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7388)
  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 7388)
  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..7388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     432..454
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     516..535
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    complement(586..619)
                     /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(670..703)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(737..1450,1451..1453,1454..1456))
                     /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(782..803)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1131..1150
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1390..1411
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1453..1462
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2514..2547
                     /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    2598..2631
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2656..3244
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2709..2729)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3127..3138)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3271..3288)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3277..3501
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        complement(3711..4040)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3725..3860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4025..4045
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(4286..4874)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4366..4385)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5045..5836,5837..5905))
                     /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     5668..5687
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5906..6010)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6078..6096
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6134..6156)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6256..6275
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6489..6508
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        6798..6816
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6798..6815
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     repeat_region   6837..6966
                     /label=AAV2 ITR
ORIGIN
        1 gcgccaggcg gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg
       61 gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg
      121 cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc
      181 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
      241 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
      301 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
      361 ttgtgcgggg ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg
      421 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca
      481 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc
      541 tcatcatttt ggcaaagaat tggatcctct agagtcgact ccggaataac ttcgtatagg
      601 atactttata cgaagttatg cagaatggta gctggattgt agctgctatt agcaatatga
      661 aacctcttaa taacttcgta tagcatacat tatacgaagt tatggcgcgc cagcgggttt
      721 aaacgggccc tctagattac ttgtacagct cgtccatgcc gagagtgatc ccggcggcgg
      781 tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg ctcagtttgg
      841 actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt
      901 tctgctggta gtggtcggcg agctgcacgc cgccgtcctc gatgttgtgg cggatcttga
      961 agttggcctt gatgccgttc ttctgcttgt cggcggtgat atagacgttg tggctgttgt
     1021 agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg atgcccttca
     1081 gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt
     1141 tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc atggcggact
     1201 tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc aggccgtagc
     1261 ccagggtggt cacgagggtg ggccagggca cgggcagctt gccggtggtg cagatcagct
     1321 tcagggtcag cttgccgtag gtggcatcgc cctcgccctc gccggacacg ctgaacttgt
     1381 ggccgtttac gtcgccgtcc agctcgacca ggatgggcac caccccggtg aacagctcct
     1441 cgcccttgct caccatggcg gccgcgctcg gggagacctg ccccggcgcg gacgccaggg
     1501 tcacgctggt ggccgacgtg gaggccgccg tcgccgactc cgaggaaggc tctgcccagg
     1561 ggttgcgtcc gcagagcagg aagggcacgg cgcagtcccg gaactggcgg ttcatgaaga
     1621 tgtagatgat ggggttgtag accgtgctcg tcttggtcag gtacatgggc agtgtggcga
     1681 tgaccgggtc gatgtacacg tcgggcttgg tcaccacgat catggcgaag agcgcgtagg
     1741 gcagccagca gacgaggaag gccaccacca tgaccaccac catgcgcgac acctgcgcct
     1801 ccgccttggc ggtgctgccg ctctcgccca tccccagctt ggccacctgg tggagggtcc
     1861 acatgaggcg gccgtacgcc accacgatga cgaggaaggg gatggcgaag cagaaggcga
     1921 agtagctcgt aatgtacgac acgttggcgg ggtcgcggct gtaccagtcg ggcgcgcacg
     1981 acgtccgcac gccctccagc tcgtagctgc cccagccgaa gagcggcggc gtgttccaca
     2041 cgaacgacca gagccaggcc caggcgatgc ccagcagcgc gtggcgccgc gtgaacatca
     2101 gcgtgcccag cggcttgcac accaccacga agcgatcgac ggcgatcacg gcgatcgtgc
     2161 acagagcggc gatgccgaag aaagcgacgg cgaacccttc gatgacgcag ccgacgcggc
     2221 cgaggttgaa gtacccgacg gcgttggtct ccacgacgag gctggcgccg aacaccgtga
     2281 cgcccaggtc cgccacggcg aggttgacga gcgagaagtt gagcgggtgg cgtagctggc
     2341 ggtggcgcag ggtgacgatg acgacggtgc tgttgagcac gagcgaggcg atggtgaaca
     2401 cggccatgat cacggcgagg atggtgaagc cgtagcgggg catcaacggg acctcgccgt
     2461 tgggcaggag gtcgagcgag gtcaagttct ccatgaattc caccggtgct agcataactt
     2521 cgtataaagt atcctatacg aagttatttg ccttaaccca gaaattatca ctgttattct
     2581 ttagaatggt gcaaagaata acttcgtata atgtatgcta tacgaagtta tgaattcgat
     2641 atcaagctta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     2701 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     2761 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     2821 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     2881 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     2941 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3001 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3061 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac
     3121 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3181 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3241 ccgcatcgat accgtcgact cgctgatcag cctcgactgt gccttctagt tgccagccat
     3301 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3361 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3421 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     3481 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tcgactagag
     3541 catggctacg tagataagta gcatggcggg ttaatcatta actacaagga acccctagtg
     3601 atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag
     3661 gtcgcccgac gcccgggcgg cctcagtgag cgagcgagcg cgcagagctt tttgcaaaag
     3721 cctaggcctc caaaaaagcc tcctcactac ttctggaata gctcagaggc cgaggcggcc
     3781 tcggcctctg cataaataaa aaaaattagt cagccatggg gcggagaatg ggcggaactg
     3841 ggcggagtta ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg
     3901 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt
     3961 gctgactaat tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt
     4021 ccacacccta actgacacac attccacagc tgcattaatg aatcggccaa cgcgcgggga
     4081 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     4141 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     4201 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     4261 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     4321 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     4381 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4441 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4501 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4561 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4621 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4681 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4741 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4801 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4861 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4921 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4981 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     5041 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     5101 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     5161 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     5221 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     5281 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     5341 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     5401 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5461 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5521 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5581 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5641 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5701 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5761 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5821 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5881 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5941 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     6001 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     6061 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     6121 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     6181 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6241 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat tcgacgctct
     6301 cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc
     6361 gccgccgcaa ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg
     6421 cctgccacca tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct
     6481 tccccatcgg tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg
     6541 ccggccacga tgcgtccggc gtagaggatc tggctagcga tgaccctgct gattggttcg
     6601 ctgaccattt ccgggtgcgg gacggcgtta ccagaaactc agaaggttcg tccaaccaaa
     6661 ccgactctga cggcagttta cgagagagat gatagggtct gcttcagtaa gccagatgct
     6721 acacaattag gcttgtacat attgtcgtta gaacgcggct acaattaata cataacctta
     6781 tgtatcatac acatacgatt taggtgacac tatagaatac acggaattaa ttctagctgc
     6841 gcgctcgctc gctcactgag gccgcccggg caaagcccgg gcgtcgggcg acctttggtc
     6901 gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg
     6961 gttccttgta gttaatgatt aacccgccat gctacttatc tacgtagcca tgctctaggt
     7021 accattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg
     7081 acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca
     7141 tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc
     7201 ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc
     7261 tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc
     7321 cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg
     7381 gggggggg
//
