LOCUS       sequence-391009-        5616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tdTomato under the control of TRE3G.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5616)
  AUTHORS   Lee JH, Kim WB, Park EH, Cho JH
  TITLE     Neocortical synaptic engrams for remote contextual memories.
  JOURNAL   Nat Neurosci. 2022 Dec 23. doi: 10.1038/s41593-022-01223-1.
  PUBMED    36564546
REFERENCE   2  (bases 1 to 5616)
  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: "Nat 
            Neurosci. 2022 Dec 23. doi: 10.1038/s41593-022-01223-1."
COMMENT     Sequence Label: pAAV-TRE3G-tdTomato
FEATURES             Location/Qualifiers
     source          1..5616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   11..140
                     /label=AAV2 ITR
     protein_bind    147..167
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    173..191
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    209..227
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    245..263
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    281..299
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    317..335
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    353..371
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    389..407
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     452..476
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    543..567
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      568..577
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             574..2004
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     protein_bind    complement(2005..2029)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2039..2627
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2092..2112)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2510..2521)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2657..2674)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2663..2870
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2878..3018
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2878..3007
                     /label=AAV2 ITR
     rep_origin      3093..3548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3180..3199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3390..3411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3565..3584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3684..3706
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3744..3762)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3830..3934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3935..4003,4004..4795)
                     /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(4153..4172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4966..5554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5455..5474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   join(5616,1..140)
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
ORIGIN
        1 ctgcaggcag ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg
       61 ggcgaccttt ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa
      121 ctccatcact aggggttcct tctagacaac tttgtataga aaagttgttt actccctatc
      181 agtgatagag aacgtatgaa gagtttactc cctatcagtg atagagaacg tatgcagact
      241 ttactcccta tcagtgatag agaacgtata aggagtttac tccctatcag tgatagagaa
      301 cgtatgacca gtttactccc tatcagtgat agagaacgta tctacagttt actccctatc
      361 agtgatagag aacgtatatc cagtttactc cctatcagtg atagagaacg tataagcttt
      421 aggcgtgtac ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct
      481 ggagcaattc cacaacactt ttgtcttata ccaactttcc gtaccacttc ctaccctcgt
      541 aaacaagttt gtacaaaaaa gcaggctgcc accatggtga gcaagggcga ggaggtcatc
      601 aaagagttca tgcgcttcaa ggtgcgcatg gagggctcca tgaacggcca cgagttcgag
      661 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      721 accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt catgtacggc
      781 tccaaggcgt acgtgaagca ccccgccgac atccccgatt acaagaagct gtccttcccc
      841 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggtctggt gaccgtgacc
      901 caggactcct ccctgcagga cggcacgctg atctacaagg tgaagatgcg cggcaccaac
      961 ttcccccccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctccaccgag
     1021 cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc accaggccct gaagctgaag
     1081 gacggcggcc actacctggt ggagttcaag accatctaca tggccaagaa gcccgtgcaa
     1141 ctgcccggct actactacgt ggacaccaag ctggacatca cctcccacaa cgaggactac
     1201 accatcgtgg aacagtacga gcgctccgag ggccgccacc acctgttcct ggggcatggc
     1261 accggcagca ccggcagcgg cagctccggc accgcctcct ccgaggacaa caacatggcc
     1321 gtcatcaaag agttcatgcg cttcaaggtg cgcatggagg gctccatgaa cggccacgag
     1381 ttcgagatcg agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg
     1441 aaggtgacca agggcggccc cctgcccttc gcctgggaca tcctgtcccc ccagttcatg
     1501 tacggctcca aggcgtacgt gaagcacccc gccgacatcc ccgattacaa gaagctgtcc
     1561 ttccccgagg gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg tctggtgacc
     1621 gtgacccagg actcctccct gcaggacggc acgctgatct acaaggtgaa gatgcgcggc
     1681 accaacttcc cccccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc
     1741 accgagcgcc tgtacccccg cgacggcgtg ctgaagggcg agatccacca ggccctgaag
     1801 ctgaaggacg gcggccacta cctggtggag ttcaagacca tctacatggc caagaagccc
     1861 gtgcaactgc ccggctacta ctacgtggac accaagctgg acatcacctc ccacaacgag
     1921 gactacacca tcgtggaaca gtacgagcgc tccgagggcc gccaccacct gttcctgtac
     1981 ggcatggacg agctgtacaa gtaaacccag ctttcttgta caaagtggga attccgataa
     2041 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
     2101 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
     2161 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
     2221 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
     2281 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
     2341 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
     2401 gggcactgac aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc
     2461 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
     2521 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
     2581 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gggaattcct
     2641 agagctcgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     2701 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     2761 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2821 caggacagca agggggagga ttgggaagag aatagcaggc atgctgggga gggccgcagg
     2881 aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg
     2941 ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag
     3001 cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta
     3061 tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc
     3121 ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct tagcgcccgc
     3181 tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct
     3241 aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa
     3301 acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc
     3361 tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact
     3421 caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt cggtctattg
     3481 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt
     3541 tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc
     3601 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     3661 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     3721 accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat
     3781 gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     3841 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     3901 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     3961 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     4021 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     4081 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     4141 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     4201 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     4261 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     4321 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     4381 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     4441 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     4501 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     4561 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     4621 tgctgataaa tctggagccg gtgagcgtgg aagccgcggt atcattgcag cactggggcc
     4681 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     4741 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     4801 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     4861 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     4921 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     4981 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5041 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5101 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5161 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5221 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5281 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5341 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5401 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     5461 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     5521 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     5581 gttcctggcc ttttgctggc cttttgctca catgtc
//
