LOCUS       sequence-497741-        6793 bp    DNA     circular SYN 16-JAN-2026
DEFINITION  Cre-dependent AAV expressing mCherry and a scrambled shRNA under 
            EF1-alpha promoter. Used as a non-targeting control for shRNA 
            knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6793)
  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 6793)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 16, 2026 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-Scramble
FEATURES             Location/Qualifiers
     source          1..6793
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1446..1479)
                     /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(1530..1563)
                     /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(1652..1672)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     CDS             complement(1772..2482)
                     /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(2050..2069)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2319..2337
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      2479..2488
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2610..2643
                     /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    2694..2727
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     2763..2782
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_feature    2920..3508
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2973..2993)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3391..3402)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3540..4016
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3739..3758)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4056..4196
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4056..4185
                     /label=AAV2 ITR
     rep_origin      4271..4726
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4358..4377)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4568..4589
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4743..4762)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4862..4884
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4922..4940)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5008..5112
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5113..5181,5182..5973)
                     /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(5331..5350)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6144..6732
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6633..6652
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc ctctagattc gctagcggat
     1441 ccggaataac ttcgtatagg atactttata cgaagttatg cagaatggta gctggattgt
     1501 agctgctatt agcaatatga aacctcttaa taacttcgta tagcatacat tatacgaagt
     1561 tattattttg tactaatatc tttgtgttta cagtcaaatt aattctaatt atctctctaa
     1621 cagccttgta tcgtatatgc aaatatgaag gaatcatggg aaataggccc tcttcctgcc
     1681 cgaccttggc gcgcgctcgg cgcgcggtca cgctccgtca cgtggtgcgt tttgcctgcg
     1741 cgtctttcca ctgggttaat taagcggccg cttacttgta cagctcgtcc atgccgccgg
     1801 tggagtggcg gccctcggcg cgttcgtact gttccacgat ggtgtagtcc tcgttgtggg
     1861 aggtgatgtc caacttgatg ttgacgttgt aggcgccggg cagctgcacg ggcttcttgg
     1921 ccttgtaggt ggtcttgacc tcagcgtcgt agtggccgcc gtccttcagc ttcagcctct
     1981 gcttgatctc gcccttcagg gcgccgtcct cggggtacat ccgctcggag gaggcctccc
     2041 agcccatggt cttcttctgc attacggggc cgtcggaggg gaagttggtg ccgcgcagct
     2101 tcaccttgta gatgaactcg ccgtcctgca gggaggagtc ctgggtcacg gtcaccacgc
     2161 cgccgtcctc gaagttcatc acgcgctccc acttgaagcc ctcggggaag gacagcttca
     2221 agtagtcggg gatgtcggcg gggtgcttca cgtaggcctt ggagccgtac atgaactgag
     2281 gggacaggat gtcccaggcg aagggcaggg ggccaccctt ggtcaccttc agcttggcgg
     2341 tctgggtgcc ctcgtagggg cggccctcgc cctcgccctc gatctcgaac tcgtggccgt
     2401 tcacggagcc ctccatgtgc accttgaagc gcatgaactc cttgatgatg gccatgttat
     2461 cctcctcgcc cttgctcacc atggtggcgg atgctaccgg tggatcccgg gcccgcggta
     2521 ccgtcgactg cagctcctga tcggcctctt cattcttctc aaccggttta gttcttctga
     2581 tgcagcacag catggggcgg cgcgccgcga taacttcgta taaagtatcc tatacgaagt
     2641 tatttgcctt aacccagaaa ttatcactgt tattctttag aatggtgcaa agaataactt
     2701 cgtataatgt atgctatacg aagttatgtt tatctgcagt tttaaaatta tgttttaaaa
     2761 tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttgggt ttatatatct
     2821 tgtggaaagg acgcggtttc ctagggagct tcccagttaa gtacctgacc catacttaac
     2881 tgggaagctc tttttgaatt cgatatcaag cttatcgata atcaacctct ggattacaaa
     2941 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     3001 gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc
     3061 ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt
     3121 ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc
     3181 tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc
     3241 gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg
     3301 gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt
     3361 ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc
     3421 cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt
     3481 cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg
     3541 ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag
     3601 tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct
     3661 tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa
     3721 cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc
     3781 tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt
     3841 tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg
     3901 gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt
     3961 ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga
     4021 ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc
     4081 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     4141 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     4201 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     4261 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     4321 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     4381 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     4441 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4501 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4561 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4621 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4681 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4741 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4801 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4861 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4921 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4981 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5041 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5101 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5161 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5221 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5281 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5341 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5401 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5461 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5521 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5581 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5641 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5701 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5761 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5821 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5881 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5941 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6001 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6061 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6121 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6181 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6241 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6301 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6361 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6421 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6481 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6541 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6601 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6661 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6721 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6781 tttgctcaca tgt
//
