LOCUS       sequence-123800-        3152 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  expresses dsRed and shRNA to knock down adra2a receptors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3152)
  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 3152)
  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-adra2a.1227.shRNA
FEATURES             Location/Qualifiers
     source          1..3152
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     32..54
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     protein_bind    170..203
                     /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    266..299
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             313..990
                     /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(443..461)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     complement(493..513)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     711..730
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     905..928
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     ncRNA           1040..1161
                     /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           1189..1203
                     /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           1231..1357
                     /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        1511..1613
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1614..2273
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1680..1699)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             2403..2432
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     protein_bind    complement(2458..2491)
                     /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(2562..2595)
                     /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(2687..2707)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 ggctgccttc gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc
       61 tctagagcct ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac
      121 gtgctggtta ttgtgctgtc tcatcatttt ggcaaagaat tggatcccca taacttcgta
      181 taaagtatcc tatacgaagt tatatcaaaa taggaagacc aatgcttcac catcgacccg
      241 aattgccaag catcaccatc gacccataac ttcgtataat gtatgctata cgaagttatg
      301 aattcctgca ggatggactc ctccgaggac gtcatcaagg agttcatgcg cttcaaggtg
      361 cgcatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      421 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggcggccc cctgcccttc
      481 gcctgggaca tcctgtcccc ccagttccag tacggctcca aggtgtacgt gaagcacccc
      541 gccgacatcc ccgactacaa gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      601 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctcctt acaggacggc
      661 tccttcatct acaaggtgaa gttcatcggc gtgaacttcc cctccgacgg ccccgtaatg
      721 cagaagaaga ctatgggctg ggaggcctcc accgagcgcc tgtacccccg cgacggcgtg
      781 ctgaagggcg agatccacaa ggccctgaag ctgaaggacg gcggccacta cctggtggag
      841 ttcaagtcca tctacatggc caagaagccc gtgcagctgc ccggctacta ctacgtggac
      901 tccaagctgg acatcacctc ccacaacgag gactacacca tcgtggagca gtacgagcgc
      961 gccgagggcc gccaccacct gttcctgtag gcggccgcaa gccttgttaa gtgctcgctt
     1021 cggcagcaca tatactatgt tgaatgaggc ttcagtactt tacagaatcg ttgcctgcac
     1081 atcttggaaa cacttgctgg gattacttct tcaggttaac ccaacagaag gctcgagaag
     1141 gtatattgct gttgacagtg agcgagcaac gtgctggtta ttatcgtagt gaagccacag
     1201 atgtacgata ataaccagca cgttgcctgc ctactgcctc ggaattcaag gggctacttt
     1261 aggagcaatt atcttgttta ctaaaactga ataccttgct atctctttga tacattttta
     1321 caaagctgaa ttaaaatggt ataaattaaa tcactttttt caattggaag actaatgcgt
     1381 ttaaacacgc ggcgacgcgt tcgaccgaat aaatacctgt gacggaagat cacttcgcag
     1441 aataaataaa tcctggtgtc cctgttgata ccgggaagcc ctgggccaac ttttggcgaa
     1501 aatgagacgt tgatcggcac gtaagaggtt ccaactttca ccataatgaa ataagatcac
     1561 taccgggcgt attttttgag ttgtcgagat tttcaggagc taaggaagct aaaatggaga
     1621 aaaaaatcac tggatatacc accgttgata tatcccaatg gcatcgtaaa gaacattttg
     1681 aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg gatattacgg
     1741 cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt attcacattc
     1801 ttgcccgcct gatgaatgct catccggaat tacgtatggc aatgaaagac ggtgagctgg
     1861 tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt
     1921 catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag
     1981 atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt
     2041 ttttcgtctc agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata
     2101 tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg
     2161 tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca
     2221 gaatgcttaa tgaattacaa cagtactgcg atgagtggca gggcggggcg taattttttt
     2281 aaggcagtta ttggtgccct taaacgcctg gttgctacgc ctgaataagt gataataagc
     2341 ggatgaatgg cagaaattcg gatctcgcgc gtttgggcgg tggctccctg ccacgcggct
     2401 ccgaacagaa gctgatctcc gaagaggatc tgacatgtgt ttaaacttaa ttaaatcata
     2461 acttcgtata ggatacttta tacgaagtta tcattgggat tcttcctatt ttgatccaag
     2521 catcaccatc gaccctctag tccagatctc accatcgacc cataacttcg tatagcatac
     2581 attatacgaa gttatgtccc tcgaagaggt tcaagcttta acagcttatc gataatcaac
     2641 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     2701 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     2761 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     2821 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     2881 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     2941 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3001 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     3061 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc tcaatccagc
     3121 ggaccttcct tcccgcggcc tgctgccggg tc
//
