LOCUS       sequence-483108-        6743 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of FlpOERT2 under the Fos promotor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6743)
  AUTHORS   Dewa KI, Kaseda K, Kuwahara A, Kubotera H, Yamasaki A, Awata N, 
            Komori A, Holtz MA, Kasai A, Skibbe H, Takata N, Yokoyama T, Tsuda 
            M, Numata G, Nakamura S, Takimoto E, Sakamoto M, Ito M, Masuda T, 
            Nagai J
  TITLE     The astrocytic ensemble acts as a multiday trace to stabilize 
            memory.
  JOURNAL   Nature. 2025 Oct 15. doi: 10.1038/s41586-025-09619-2.
  PUBMED    41094146
REFERENCE   2  (bases 1 to 6743)
  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"; journalName: "Nature. 
            2025 Oct 15. doi: 10.1038/s41586-025-09619-2."
COMMENT     Sequence Label: Fos-FlpOERT2
FEATURES             Location/Qualifiers
     source          1..6743
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     protein_bind    137..157
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             complement(857..868)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    868..892
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2180..3115
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             2228..3121
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATA"
     protein_bind    complement(3122..3146)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    3156..3744
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3209..3229)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3627..3638)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3774..3791)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3780..3987
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3995..4135
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3995..4124
                     /label=AAV2 ITR
     rep_origin      4210..4665
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4297..4316)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4507..4528
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4682..4701)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4801..4823
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4861..4879)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4947..5051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5052..5120,5121..5912)
                     /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(5270..5289)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6083..6671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6572..6591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(6733..6743,1..130)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tctagacaac tttgtataga aaagttgagc tttcctttag gaacagaggc
      181 ttcgagcctt taaggctgcg tacttgcttc tcctaatacc agagactcaa aaaaaaaaaa
      241 aaagttccag attgctggac aatgacccgg gtctcatccc ttgaccctgg gaaccgggtc
      301 cacattgaat caggtgcgaa tgttcgctcg ccttctctgc ctttcccgcc tcccctcccc
      361 cggccgcggc cccggttccc cccctgcgct gcaccctcag agttggctgc agccggcgag
      421 ctgttcccgt caatccctcc ctcctttaca caggatgtcc atattaggac atctgcgtca
      481 gcaggtttcc acggccggtc cctgttgttc tggggggggg accatctccg aaatcctaca
      541 cgcggaaggt ctaggagacc ccctaagatc ccaaatgtga acactcatag gtgaaagatg
      601 tatgccaaga cgggggttga aagcctgggg cgtagagttg acgacagagc gcccgcagag
      661 ggccttgggg cgcgcttccc cccccttcca gttccgccca gtgacgtagg aagtccatcc
      721 attcacagcg cttctataaa ggcgccagct gaggcgccta ctactccaac cgcgactgca
      781 gcgagcaact gagaagactg gatagagccg gcggttccgc gaacgagcag tgaccgcgct
      841 cccacccagc tctgctctgc agctcccaca agtttgtaca aaaaagcagg ctgccaccat
      901 gagccagttc gacatcctgt gcaagacccc ccccaaggtg ctggtgcggc agttcgtgga
      961 gagattcgag aggcccagcg gcgagaagat cgccagctgt gccgccgagc tgacctacct
     1021 gtgctggatg atcacccaca acggcaccgc catcaagagg gccaccttca tgagctacaa
     1081 caccatcatc agcaacagcc tgagcttcga catcgtgaac aagagcctgc agttcaagta
     1141 caagacccag aaggccacca tcctggaggc cagcctgaag aagctgatcc ccgcctggga
     1201 gttcaccatc atcccttaca acggccagaa gcaccagagc gacatcaccg acatcgtgtc
     1261 cagcctgcag ctgcagttcg agagcagcga ggaggccgac aagggcaaca gccacagcaa
     1321 gaagatgctg aaggccctgc tgtccgaggg cgagagcatc tgggagatca ccgagaagat
     1381 cctgaacagc ttcgagtaca ccagcaggtt caccaagacc aagaccctgt accagttcct
     1441 gttcctggcc acattcatca actgcggcag gttcagcgac atcaagaacg tggaccccaa
     1501 gagcttcaag ctggtgcaga acaagtacct gggcgtgatc attcagtgcc tggtgaccga
     1561 gaccaagaca agcgtgtcca ggcacatcta ctttttcagc gccagaggca ggatcgaccc
     1621 cctggtgtac ctggacgagt tcctgaggaa cagcgagccc gtgctgaaga gagtgaacag
     1681 gaccggcaac agcagcagca acaagcagga gtaccagctg ctgaaggaca acctggtgcg
     1741 cagctacaac aaggccctga agaagaacgc cccctacccc atcttcgcta tcaagaacgg
     1801 ccctaagagc cacatcggca ggcacctgat gaccagcttt ctgagcatga agggcctgac
     1861 cgagctgaca aacgtggtgg gcaactggag cgacaagagg gcctccgccg tggccaggac
     1921 cacctacacc caccagatca ccgccatccc cgaccactac ttcgccctgg tgtccaggta
     1981 ctacgcctac gaccccatca gcaaggagat gatcgccctg aaggacgaga ccaaccccat
     2041 cgaggagtgg cagcacatcg agcagctgaa gggcagcgcc gagggcagca tcagataccc
     2101 cgcctggaac ggcatcatca gccaggaggt gctggactac ctgagcagct acatcaacag
     2161 gcggatcctc gagccatctg ctggagacat gagagctgcc aacctttggc caagcccgct
     2221 catgatcaaa cgctctaaga agaacagcct ggccttgtcc ctgacggccg accagatggt
     2281 cagtgccttg ttggatgctg agccccccat actctattcc gagtatgatc ctaccagacc
     2341 cttcagtgaa gcttcgatga tgggcttact gaccaacctg gcagacaggg agctggttca
     2401 catgatcaac tgggcgaaga gggtgccagg ctttgtggat ttgaccctcc atgatcaggt
     2461 ccaccttcta gaatgtgcct ggctagagat cctgatgatt ggtctcgtct ggcgctccat
     2521 ggagcaccca gtgaagctac tgtttgctcc taacttgctc ttggacagga accagggaaa
     2581 atgtgtagag ggcatggtgg agatcttcga catgctgctg gctacatcat ctcggttccg
     2641 catgatgaat ctgcagggag aggagtttgt gtgcctcaaa tctattattt tgcttaattc
     2701 tggagtgtac acatttctgt ccagcaccct gaagtctctg gaagagaagg accatatcca
     2761 ccgagtcctg gacaagatca cagacacttt gatccacctg atggccaagg caggcctgac
     2821 cctgcagcag cagcaccagc ggctggccca gctcctcctc atcctctccc acatcaggca
     2881 catgagtaac aaaggcatgg agcatctgta cagcatgaag tgcaagaacg tggtgcccct
     2941 ctatgacctg ctgctggagg cggcggacgc ccaccgccta catgcgccca ctagccgtgg
     3001 aggggcatcc gtggaggaga cggaccaaag ccacttggcc actgcgggct ctacttcatc
     3061 gcattccttg caaaagtatt acatcacggg ggaggcagag ggtttccctg ccacagcttg
     3121 aacccagctt tcttgtacaa agtgggaatt ccgataatca acctctggat tacaaaattt
     3181 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3241 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3301 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3361 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3421 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3481 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3541 tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc
     3601 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3661 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3721 tctccctttg ggccgcctcc ccgcatcggg aattcctaga gctcgctgat cagcctcgac
     3781 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     3841 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     3901 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     3961 ggaagagaat agcaggcatg ctggggaggg ccgcaggaac ccctagtgat ggagttggcc
     4021 actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc
     4081 ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc
     4141 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat acgtcaaagc
     4201 aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     4261 gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc ttcccttcct
     4321 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4381 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt gatggttcac
     4441 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4501 ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg ggctattctt
     4561 ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag ctgatttaac
     4621 aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg tgcactctca
     4681 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4741 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4801 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4861 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4921 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4981 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5041 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5101 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5161 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5221 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5281 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5341 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5401 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5461 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5521 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5581 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5641 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5701 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5761 agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5821 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5881 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5941 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6001 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6061 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6721 ttgctcacat gtcctgcagg cag
//
