LOCUS       sequence-121284-        3107 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  AAV vector to express dsRed sh RNA (scramble).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3107)
  AUTHORS   Zhang Z, Ferretti V, Guntan I, Moro A, Steinberg EA, Ye Z, Zecharia 
            AY, Yu X, Vyssotski AL, Brickley SG, Yustos R, Pillidge ZE, Harding 
            EC, Wisden W, Franks NP
  TITLE     Neuronal ensembles sufficient for recovery sleep and the sedative 
            actions of alpha2 adrenergic agonists.
  JOURNAL   Nat Neurosci. 2015 Apr;18(4):553-61. doi: 10.1038/nn.3957. Epub 2015
            Feb 23.
  PUBMED    25706476
REFERENCE   2  (bases 1 to 3107)
  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: "10.1038/nn.3957"; 
            journalName: "Nat Neurosci"; date: "2015-04"; volume: "18"; issue: 
            "4"; pages: "553-61"
COMMENT     Sequence Label: AAV-actin prom-dsRed-scramble shRNA
FEATURES             Location/Qualifiers
     source          1..3107
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     74..93
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     protein_bind    128..161
                     /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    224..257
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             271..948
                     /codon_start=1
                     /product="rapidly maturing tetrameric variant of DsRed 
                     fluorescent protein (Bevis and Glick, 2002)"
                     /label=DsRed-Express
                     /note="mammalian codon-optimized"
                     /translation="MDSSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGSFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGEIHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDSKLDITSHNEDYTIVEQYERAEGRHH
                     LFL"
     primer_bind     complement(401..419)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(451..471)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     669..688
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     863..886
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     ncRNA           998..1119
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1146..1160
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           1187..1313
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     promoter        1466..1568
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1569..2228
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1635..1654)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2358..2387
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(2413..2446)
                     /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(2517..2550)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2642..2662)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 ctggcggtga ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc
       61 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaattg
      121 gatccccata acttcgtata aagtatccta tacgaagtta tatcaaaata ggaagaccaa
      181 tgcttcacca tcgacccgaa ttgccaagca tcaccatcga cccataactt cgtataatgt
      241 atgctatacg aagttatgaa ttcctgcagg atggactcct ccgaggacgt catcaaggag
      301 ttcatgcgct tcaaggtgcg catggagggc tccgtgaacg gccacgagtt cgagatcgag
      361 ggcgagggcg agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag
      421 ggcggccccc tgcccttcgc ctgggacatc ctgtcccccc agttccagta cggctccaag
      481 gtgtacgtga agcaccccgc cgacatcccc gactacaaga agctgtcctt ccccgagggc
      541 ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac
      601 tcctccttac aggacggctc cttcatctac aaggtgaagt tcatcggcgt gaacttcccc
      661 tccgacggcc ccgtaatgca gaagaagact atgggctggg aggcctccac cgagcgcctg
      721 tacccccgcg acggcgtgct gaagggcgag atccacaagg ccctgaagct gaaggacggc
      781 ggccactacc tggtggagtt caagtccatc tacatggcca agaagcccgt gcagctgccc
      841 ggctactact acgtggactc caagctggac atcacctccc acaacgagga ctacaccatc
      901 gtggagcagt acgagcgcgc cgagggccgc caccacctgt tcctgtaggc ggccgcaagc
      961 cttgttaagt gctcgcttcg gcagcacata tactatgttg aatgaggctt cagtacttta
     1021 cagaatcgtt gcctgcacat cttggaaaca cttgctggga ttacttcttc aggttaaccc
     1081 aacagaaggc tcgagaaggt atattgctgt tgacagtgag cggccgcgat taggctgtta
     1141 taatagtgaa gccacagatg tattataaca gcctaatcgc ggctgcctac tgcctcggaa
     1201 ttcaaggggc tactttagga gcaattatct tgtttactaa aactgaatac cttgctatct
     1261 ctttgataca tttttacaaa gctgaattaa aatggtataa attaaatcac ttttttcaat
     1321 tggaagacta atgcgtttaa acacgcggcg acgcgttcga ccgaataaaa cctgtgacgg
     1381 aagatcactt cgcagaataa ataaatcctg gtgtccctgt tgataccggg aagccctggg
     1441 ccaacttttg gcgaaaatga gacgttgatc ggcacgtaag aggttccaac tttcaccata
     1501 atgaaataag atcactaccg ggcgtatttt ttgagttgtc gagattttca ggagctaagg
     1561 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     1621 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     1681 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     1741 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattacgt atggcaatga
     1801 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     1861 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     1921 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     1981 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     2041 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     2101 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     2161 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     2221 gggcgtaatt tttttaaggc agttattggt gcccttaaac gcctggttgc tacgcctgaa
     2281 taagtgataa taagcggatg aatggcagaa attcggatct cgcgcgtttg ggcggtggct
     2341 ccctgccacg cggctccgaa cagaagctga tctccgaaga ggatctgaca tgtgtttaaa
     2401 cttaattaaa tcataacttc gtataggata ctttatacga agttatcatt gggattcttc
     2461 ctattttgat ccaagcatca ccatcgaccc tctagtccag atctcaccat cgacccataa
     2521 cttcgtatag catacattat acgaagttat gtccctcgaa gaggttcaag ctttaacagc
     2581 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2641 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2701 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2761 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2821 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2881 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2941 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     3001 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     3061 cggcctcaat ccagcggacc ttccttcccg cggcctgctg ccgggtc
//
