LOCUS       sequence-132478-        7746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This plasmid is pAAV-panpromoter-flex-rev-gamma2F77-2A-Venus. 
            Confers zolpidem sensitivity on zolpidem-insensitive neurons..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7746)
  AUTHORS   Uygun DS, Ye Z, Zecharia AY, Harding EC, Yu X, Yustos R, Vyssotski 
            AL, Brickley SG, Franks NP, Wisden W
  TITLE     Bottom-Up versus Top-Down Induction of Sleep by Zolpidem Acting on 
            Histaminergic and Neocortex Neurons.
  JOURNAL   J Neurosci. 2016 Nov 2;36(44):11171-11184.
  PUBMED    27807161
REFERENCE   2  (bases 1 to 7746)
  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: "J 
            Neurosci."; date: "2016-11-2"; volume: "36(44)"; pages: 
            "11171-11184"
COMMENT     Sequence Label: pAAV-flex-rev-g2-2A-Venus
FEATURES             Location/Qualifiers
     source          1..7746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    complement(1359..1392)
                     /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(1443..1476)
                     /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(1483..2196,2197..2199,2200..2202))
                     /codon_start=1
                     /product="yellow fluorescent protein (YFP) with fast and 
                     efficient maturation (Nagai et al., 2002)"
                     /label=Venus
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1528..1549)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1877..1896
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2136..2157
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     CDS             complement(2218..2271)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     protein_bind    3731..3764
                     /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    3815..3848
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    3873..4461
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3926..3946)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4344..4355)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4493..4969
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4692..4711)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5009..5149
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5009..5138
                     /label=AAV2 ITR
     rep_origin      5224..5679
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5311..5330)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5521..5542
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5696..5715)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5815..5837
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5875..5893)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5961..6065
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6066..6134,6135..6926)
                     /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(6284..6303)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7097..7685
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7586..7605
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtgacca ccccccatga agtctgttgt
      181 gtaagtacag gtaatgcttt agacactgtt tttctcagat tgtgttcaag ctcttgggtg
      241 caggcatccg cagtccagag cctctgagga acaaagggac ctcgatataa agaacaaaaa
      301 gctcttggta ttcgggaaaa ctccgaagga tttttgttgt tttgtggcag tgttggggtt
      361 caggacaaac acaccacccc agaatatgat ttcaggagaa actgccatcc ccgcccccac
      421 tagtacgctt cctttgtgtt atctcgagaa gccctggacg taggaggagt tctcaaaagc
      481 tgttctttcg taagagatgg tcaggaagaa acgttgatta ggaaaagtac tcgactcagg
      541 aagaggaggg gtacagataa ctgctgtcac cagaaagatt tccatctgac aaagtggcaa
      601 ttcttccccc ttacgaatgt gtgtcccgca ctctgctgca cccaggctcc gctctgttct
      661 ccagctctgc ttgctgtaga agcgtccatc cactaaaggg acttccagat ttattggcac
      721 tgaacaggtt tttcctccgt tagcttaaac agtagtcgag gcaaagaacc cagaacaagt
      781 gctatgtagt aatggataaa aagaatcctt catatcctgt tgttgatctg tcctgagctc
      841 agcctggcta gctatgggct tctctgagcc ggctctgccc tcacctcata gagaagccag
      901 ggcaggagca actccaccct ctgtgctact tattgtgaat gcccttggcc tatttctggc
      961 ctctgagcta ggagctttat tcaaggctgg ggctctatgg tctgaatgca ggggcaggga
     1021 tgatagaggg ctcttttacc ttgtctatcc agagctgctt gtaggcttgg gaggctaggt
     1081 gccccaactg gtctaaaaac accatgttct ctgattctgt tgatcaaagt tcaaatcaga
     1141 aaggacgggg acaagagagg ctgaaattaa gcctgaagga agggacttta tgggggcggg
     1201 gctaaaggag gggaataggg aactccttaa ataagagatg cactggctgc cagagagtgc
     1261 acagcacaga caaaggcgac caagaagccc tgttgacagc tgccttccag cctcctctgt
     1321 ctgtctgcca ggaggagcaa tccaaggtcg actccggaat aacttcgtat aggatacttt
     1381 atacgaagtt atgcagaatg gtagctggat tgtagctgct attagcaata tgaaacctct
     1441 taataacttc gtatagcata cattatacga agttatcgcg ccttacttgt acagctcgtc
     1501 catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt gatcgcgctt
     1561 ctcgttgggg tctttgctca gggcggactg gtagctcagg tagtggttgt cgggcagcag
     1621 cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct gcacgccgcc
     1681 gtcctcgatg ttgtggcgga tcttgaagtt ggccttgatg ccgttcttct gcttgtcggc
     1741 ggtgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca ggatgttgcc
     1801 gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt cgccctcgaa
     1861 cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg tgcgctcctg
     1921 gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt ggtcggggta
     1981 gcgggcgaag cactgcaggc cgtagcccag ggtggtcacg agggtgggcc agggcacggg
     2041 cagcttgccg gtggtgcaga tcagcttcag ggtcagcttg ccgtaggtgg catcgccctc
     2101 gccctcgccg gacacgctga acttgtggcc gtttacgtcg ccgtccagct cgaccaggat
     2161 gggcaccacc ccggtgaaca gctcctcgcc cttgctcacc atggtggatc ccggtgcagg
     2221 gccgggattc tcctccacgt caccgcatgt tagaagactt cctctgccct cgagcaaacg
     2281 tttcgcacgc agataaagat aggagaccca gtaaacaaga ttgaacaagc aaaaggcggt
     2341 agggaagaag atccgagcat aggagtccat tttggcaatg cgaatatgta tcctcccatg
     2401 tctccaggct cctgttcggc aatcttcaaa acagcagaag aaactggcac agtccttgcc
     2461 atccaaacac tcatagccat attcttcatc cctctcttga aggtgtgtgg cattgttcat
     2521 ttgaatggtt gctgatctgg gacgaatatc aatggtaggg gccttgaagg aaaacatccg
     2581 aagaagaggg tttttcttct ttttgtcttt atccttgctt ggcttccggt tgctgacaaa
     2641 ataatgcagg gtgccatact ccaccaaagc agaaaacaca aagatgaagc aaacagatac
     2701 aaagagatcc attgctgtga cataggagac cttgggcaga gattttctgg ctatggtgct
     2761 taaagttgtc atggtcagga cagtagtgat tcctaaagat gttctggcag gaacagcatc
     2821 cttattgatc cagaaggaca cccaggacag gaccacgatg agtgtgcagg gaatgtaagt
     2881 ctggatggtg aagtagccca ttcttctgct cagatcgaag tacacagaca tcaccacata
     2941 gtcaccagaa gttgtcttca ctacttcagt tgtattcctc aatccaacaa aggaaaattg
     3001 atacagcctc catgatcttg tgtcacccac ttcaacagaa ctgcgcttcc attgataaac
     3061 aatttcttca cgaggatatc cataactgga gaactccagg gggcaggagt gttcatccat
     3121 tgggaagttg tgtaattgca actggcactc agcatcaatt gttagcctta aggtgtagag
     3181 aactcgacca tcattccaaa ttctcagcat cctgttggga gtggtgatcc agtgagcatc
     3241 agcctttttg gagttcctga agaaagtgtc tggaatccag attttcccca ccatattgct
     3301 attcaaccgg agaactttaa tggtactgtt aaatttcaaa cgtctgtcat accaggtttg
     3361 ggcaaaaaaa atatcaattg tatattccat attgatagca ttcactggac caatgctgtt
     3421 cacatacata tctgtatgaa ttaatgttgg tttcactccg atgtcaggtc gaagtttgtt
     3481 gtcatagcct tccagcaggt tgtttaagat gacagtgaca tcaccctctg gaacttttgg
     3541 agtcaacacc catgttttat tagaagtgta atcttcatag tcatcgatgt acctgtaggt
     3601 gtcatctgac ttttggcttg tgaagcctgg gtagagcgat agcaggagca gaatccacag
     3661 cgtcatggat ccatatattc tccctttggg ccgcctcccc gcatcgatac cgtcgacgcg
     3721 gccgcgcgga ataacttcgt ataaagtatc ctatacgaag ttatttgcct taacccagaa
     3781 attatcactg ttattcttta gaatggtgca aagaataact tcgtataatg tatgctatac
     3841 gaagttatga attcgatatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     3901 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3961 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     4021 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     4081 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     4141 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     4201 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     4261 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     4321 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     4381 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     4441 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
     4501 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     4561 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     4621 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     4681 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     4741 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     4801 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     4861 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     4921 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     4981 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     5041 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     5101 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     5161 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     5221 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     5281 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     5341 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     5401 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     5461 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     5521 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat tcttttgatt
     5581 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     5641 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     5701 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     5761 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     5821 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     5881 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5941 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     6001 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     6061 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6121 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6181 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6241 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6301 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6361 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6421 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6481 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6541 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6601 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6661 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6721 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6781 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6841 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6901 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6961 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7021 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7081 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7141 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7201 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt
     7261 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7321 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7381 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7441 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7501 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7561 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7621 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7681 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7741 acatgt
//
