LOCUS       sequence-358660-        6817 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector for expressing CFL-SN.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6817)
  AUTHORS   Goto A, Bota A, Miya K, Wang J, Tsukamoto S, Jiang X, Hirai D, 
            Murayama M, Matsuda T, McHugh TJ, Nagai T, Hayashi Y
  TITLE     Stepwise synaptic plasticity events drive the early phase of memory 
            consolidation.
  JOURNAL   Science. 2021 Nov 12;374(6569):857-863. doi: 
            10.1126/science.abj9195. Epub 2021 Nov 11.
  PUBMED    34762472
REFERENCE   2  (bases 1 to 6817)
  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: 
            "10.1126/science.abj9195"; journalName: "Science"; date: 
            "2021-11-12- 12"; volume: "374"; issue: "6569"; pages: "857-863"
COMMENT     Sequence Label: pAAV-EF1-alpha-DIO-CFL-SN
FEATURES             Location/Qualifiers
     source          1..6817
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   2..131
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        232..1410
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          463..1401
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1358..1378
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1441..1474)
                     /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(1525..1558)
                     /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(1567..2277,2278..2283,2284..2286))
                     /codon_start=1
                     /product="monomeric variant of the photosensitizing red 
                     fluorescent protein KillerRed (Takemoto et al., 2013)"
                     /label=SuperNova
                     /translation="MGSEVGPALFQSDMTFKIFIDGEVNGQKFTIVADGSSKFPHGDFN
                     VHAVCETGKLPMSWKPICHLIQYGEPFFARYPDGISHFAQECFPEGLSIDRTVRFENDG
                     TMTSHHTYELDDTCVVSRITVNCDGFQPDGPIMRDQLVDILPSETHMFPHGPNAVRQTA
                     TIGFTTADGGKMMGHFDSKMTFNGSRAIEIPGPHFVTIITKQTRDTSDKRDHVCQREVA
                     YAHSVPRITSAIGSDED"
     protein_bind    2803..2836
                     /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    2887..2920
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2945..3533
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2998..3018)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3416..3427)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3565..4041
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3764..3783)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4081..4221
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4081..4210
                     /label=AAV2 ITR
     rep_origin      4296..4751
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4383..4402)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4593..4614
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4768..4787)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4887..4909
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4947..4965)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5033..5137
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5138..5206,5207..5998)
                     /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(5356..5375)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6169..6757
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6658..6677
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcgtc gggcgacctt
       61 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      121 taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt ttaaacagag
      181 aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt
      241 gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc
      301 ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg
      361 tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc
      421 gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt
      481 tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc
      541 tggctgcagt acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg
      601 aggccttgcg cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc
      661 tggggccgcc gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag
      721 tctctagcca tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt
      781 cttgtaaatg cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc
      841 gacggggccc gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca
      901 ccgagaatcg gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg
      961 ccgccgtgta tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga
     1021 gcggaaagat ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc
     1081 tcgggagagc gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc
     1141 gtcgcttcat gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg
     1201 agcttttgga gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc
     1261 cacactgagt gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg
     1321 gaatttgccc tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa
     1381 agtttttttc ttccatttca ggtgtcgtga ggtaccggat cctctagagt cgactccgga
     1441 ataacttcgt ataggatact ttatacgaag ttatgcagaa tggtagctgg attgtagctg
     1501 ctattagcaa tatgaaacct cttaataact tcgtatagca tacattatac gaagttatgg
     1561 cgcgccttaa tcctcgtcgc taccgatggc gctggtgatg cggggcacgc tgtgggcgta
     1621 ggccacctcg cgctggcaca cgtggtcgcg cttgtcgcta gtgtccctcg tctgcttggt
     1681 gatgatggtc acgaagtgtg ggccggggat ctcgatggcg cggctgccgt tgaaggtcat
     1741 cttgctgtcg aagtggccca tcatcttgcc gccgtcggcg gtggtgaagc cgatggtggc
     1801 ggtctggcgc acggcgttgg ggccgtgggg gaacatgtgg gtctcgctgg gcaggatgtc
     1861 caccagctgg tcgcgcatga tggggccgtc gggctggaag ccgtcgcagt tcacggtgat
     1921 gcggctcacc acgcaggtgt cgtccagctc gtaggtgtgg tggctggtca tggtgccgtc
     1981 gttctcgaag cgcacggtgc ggtcgatgct caggccctcg gggaagcact cctgggcgaa
     2041 atggctgatg ccgtcggggt agcgggcgaa gaagggctcg ccgtactgga tcaggtggca
     2101 gatgggcttc cagctcatgg gcagcttgcc ggtctcgcac acggcgtgca cgttgaagtc
     2161 gccgtggggg aacttgctgc tgccgtcggc cacgatggtg aacttctggc cgttcacctc
     2221 gccgtcgatg aagattttga aggtcatgtc gctctggaac agggcggggc cgacctctga
     2281 acccattcta gacaaaggct tgccctccag ggagatgacg gcactgcccc ccagcttctc
     2341 tgccagggtg cagcggtcct tgacctcctc gtagcagttt gcttgcaatt catgcttgat
     2401 ccctgtcagc ttcttcttga tggcgtcctt ggagctggca taaatcattt tgctcttaag
     2461 gggcgcagac tcgggggccc agaagataaa caccagatcc tccttcttgc tctccttggt
     2521 ctcataggtt gcatcataga gggcatagcg gcagtcctta tctggcagca tcttgacaaa
     2581 ggtggcgtag ggatcgtcga cagtctggcc cacatcgccc accaggatct ccttgccctc
     2641 ctccaggatg atgttcttct tgtcctcact caggcagaag agcaccgcct tcttgcgctt
     2701 cttcacctcc tctggcgttg aagacttacg caccttcatg tcgttgaaca ccttgatgac
     2761 accatcagag acagccacac cggaggccat gtttccgcta gcataacttc gtataaagta
     2821 tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt tagaatggtg
     2881 caaagaataa cttcgtataa tgtatgctat acgaagttat gaattcgata tcaagcttat
     2941 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3001 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3061 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3121 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3181 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3241 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3301 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     3361 gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3421 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3481 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3541 ccgagcgctg ctcgagagat ctacgggtgg catccctgtg acccctcccc agtgcctctc
     3601 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3661 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3721 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3781 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3841 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3901 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3961 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     4021 actgctccct tccctgtcct tctgattttg taggtaacca cgtgcggacc gagcggccgc
     4081 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     4141 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     4201 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     4261 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     4321 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     4381 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     4441 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     4501 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     4561 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     4621 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     4681 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     4741 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4801 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4861 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4921 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4981 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     5041 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5101 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5161 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5221 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5281 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5341 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5401 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5461 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5521 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5581 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5641 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5701 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5761 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5821 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5881 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5941 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     6001 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6061 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6121 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6181 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6241 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6301 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6361 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6421 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6481 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6541 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6601 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6661 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6721 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6781 acggttcctg gccttttgct ggccttttgc tcacatg
//
