LOCUS       sequence-493856-        3316 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent AAV expressing mCherry and an shRNA targeting Numb 
            under EF1-alpha promoter for selective Numb knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3316)
  AUTHORS   Shin J, Oh H, Park JH, Bae H, Yang CM, Lee M, Kim S, Heo WD
  TITLE     Dysregulation of FGFR1 signaling in the hippocampus facilitates 
            depressive disorder.
  JOURNAL   Exp Mol Med. 2025 Aug;57(8):1818-1836. doi: 
            10.1038/s12276-025-01519-9. Epub 2025 Aug 15.
  PUBMED    40813470
REFERENCE   2  (bases 1 to 3316)
  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/s12276-025-01519-9"; journalName: "Exp Mol Med"; date: 
            "2025-08"; volume: "57"; issue: "8"; pages: "1818-1836"
COMMENT     Sequence Label: AAV-EF1-alpha-DIO-DSE-mCherry-PSE-shRNA-Numb
FEATURES             Location/Qualifiers
     source          1..3316
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        2..1180
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          233..1171
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1128..1148
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1217..1250)
                     /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(1301..1334)
                     /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(1423..1443)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(1543..2253)
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1821..1840)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2090..2108
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2250..2259
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2381..2414
                     /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    2465..2498
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2534..2553
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2693..3281
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2746..2766)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3164..3175)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
       61 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      121 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      181 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      241 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      301 tacttccacc tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg
      361 ggagagttcg aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg
      421 gcctgggcgc tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc
      481 tttcgataag tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg
      541 gcaagatagt cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc
      601 cgcgggcggc gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg
      661 agcgcggcca ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc
      721 tggcctcgcg ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc
      781 agttgcgtga gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag
      841 gacgcggcgc tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc
      901 gtcctcagcc gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga
      961 ttagttctcg agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat
     1021 ggagtttccc cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta
     1081 attctccttg gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac
     1141 agtggttcaa agtttttttc ttccatttca ggtgtcgtga ggtaccggat cctctagatt
     1201 cgctagcgga tccggaataa cttcgtatag gatactttat acgaagttat gcagaatggt
     1261 agctggattg tagctgctat tagcaatatg aaacctctta ataacttcgt atagcataca
     1321 ttatacgaag ttattatttt gtactaatat ctttgtgttt acagtcaaat taattctaat
     1381 tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg gaaataggcc
     1441 ctcttcctgc ccgaccttgg cgcgcgctcg gcgcgcggtc acgctccgtc acgtggtgcg
     1501 ttttgcctgc gcgtctttcc actgggttaa ttaagcggcc gcttacttgt acagctcgtc
     1561 catgccgccg gtggagtggc ggccctcggc gcgttcgtac tgttccacga tggtgtagtc
     1621 ctcgttgtgg gaggtgatgt ccaacttgat gttgacgttg taggcgccgg gcagctgcac
     1681 gggcttcttg gccttgtagg tggtcttgac ctcagcgtcg tagtggccgc cgtccttcag
     1741 cttcagcctc tgcttgatct cgcccttcag ggcgccgtcc tcggggtaca tccgctcgga
     1801 ggaggcctcc cagcccatgg tcttcttctg cattacgggg ccgtcggagg ggaagttggt
     1861 gccgcgcagc ttcaccttgt agatgaactc gccgtcctgc agggaggagt cctgggtcac
     1921 ggtcaccacg ccgccgtcct cgaagttcat cacgcgctcc cacttgaagc cctcggggaa
     1981 ggacagcttc aagtagtcgg ggatgtcggc ggggtgcttc acgtaggcct tggagccgta
     2041 catgaactga ggggacagga tgtcccaggc gaagggcagg gggccaccct tggtcacctt
     2101 cagcttggcg gtctgggtgc cctcgtaggg gcggccctcg ccctcgccct cgatctcgaa
     2161 ctcgtggccg ttcacggagc cctccatgtg caccttgaag cgcatgaact ccttgatgat
     2221 ggccatgtta tcctcctcgc ccttgctcac catggtggcg gatgctaccg gtggatcccg
     2281 ggcccgcggt accgtcgact gcagctcctg atcggcctct tcattcttct caaccggttt
     2341 agttcttctg atgcagcaca gcatggggcg gcgcgccgcg ataacttcgt ataaagtatc
     2401 ctatacgaag ttatttgcct taacccagaa attatcactg ttattcttta gaatggtgca
     2461 aagaataact tcgtataatg tatgctatac gaagttatgt ttatctgcag ttttaaaatt
     2521 atgttttaaa atggactatc atatgcttac cgtaacttga aagtatttcg atttcttggg
     2581 tttatatatc ttgtggaaag gacgcggttt cctaggtctg tgaagatgcc gtaaacctga
     2641 cccatttacg gcatcttcac agatttttga attcgatatc aagcttatcg ataatcaacc
     2701 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2761 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2821 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2881 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2941 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     3001 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     3061 tgacaattcc gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt
     3121 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     3181 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     3241 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct
     3301 cgagagatct acgggt
//
