LOCUS       sequence-209574-        5725 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses yellow pre-eGRASP(p30) under the tetO 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5725)
  AUTHORS   Choi JH, Sim SE, Kim JI, Choi DI, Oh J, Ye S, Lee J, Kim T, Ko HG, 
            Lim CS, Kaang BK
  TITLE     Interregional synaptic maps among engram cells underlie memory 
            formation.
  JOURNAL   Science. 2018 Apr 27;360(6387):430-435. doi: 
            10.1126/science.aas9204.
  PUBMED    29700265
REFERENCE   2  (bases 1 to 5725)
  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"; 
            journalName: "Science"; date: "2018-04-27- 27"; volume: "360"; 
            issue: "6387"; pages: "430-435"
COMMENT     Sequence Label: pAAV-TIWB-yellow pre-eGRASP(p30)
FEATURES             Location/Qualifiers
     source          1..5725
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        157..535
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    165..183
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    201..219
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    237..255
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    273..291
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    309..327
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    345..363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    381..399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     444..468
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          583..715
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     698..717
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        760..778
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      782..791
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             866..1324
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKRHDFFKSAMPEGYVQERTISFKDDGK
                     YKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITA"
     primer_bind     complement(905..926)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2155..2743
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2208..2228)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2626..2637)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2756..2980
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2988..3128
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2988..3117
                     /label=AAV2 ITR
     rep_origin      3203..3658
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3290..3309)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3500..3521
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3675..3694)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3794..3816
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3854..3872)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3940..4044
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4045..4113,4114..4905)
                     /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(4263..4282)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5076..5664
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5565..5584
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgttggct ttactcccta tcagtgatag
      181 agaacgtatg aagagtttac tccctatcag tgatagagaa cgtatgcaga ctttactccc
      241 tatcagtgat agagaacgta taaggagttt actccctatc agtgatagag aacgtatgac
      301 cagtttactc cctatcagtg atagagaacg tatctacagt ttactcccta tcagtgatag
      361 agaacgtata tccagtttac tccctatcag tgatagagaa cgtataagct ttaggcgtgt
      421 acggtgggcg cctataaaag cagagctcgt ttagtgaacc gtcagatcgc ctggagcaat
      481 tccacaacac ttttgtctta taccaacttt ccgtaccact tcctaccctc gtaaagctag
      541 tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat caaggttaca
      601 agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag actcttgcgt
      661 ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc cacaggtgtc
      721 cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc actataggct
      781 agccaccatg gagaccgaca ccctcctgct atgggtgctg ctgctctggg tgcccggcag
      841 caccggcgac gcaccggtcg gaggcagcaa gggcgaggag ctgttcaccg gggtggtgcc
      901 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtga ggggcgaggg
      961 cgagggcgat gccaccatcg gcaagctgac cctgaagttc atctgcacca ccggcaagct
     1021 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcgcccg
     1081 ctaccccgac cacatgaagc ggcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1141 ccaggagcgc accatctcct tcaaggacga cggcaagtac aagacccgcg ccgtggtgaa
     1201 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcaccgact tcaaggagga
     1261 cggcaacatc ctggggcaca agctggagta caacttcaac agccacaacg tctatatcac
     1321 cgccgacaag cagaagaacg gcatcaaggc caacttcacc gtgcgccaca acgtggagga
     1381 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1441 gctgctgccc gacaaccact acctgagcta ccagaccgtg ctgagcaaag accccaacga
     1501 gaagaccggt ggatctggag gcagtggcgg atctagagct cctacaaagc ctccaccact
     1561 gccacctgga ggaggttccg gaggcggaag cggtaccgaa gtgccttcct ctatgacaac
     1621 tgagtcgaca gccactgcca tgcagtccga gatgtccacc tcaatcatgg agaccaccac
     1681 aaccctggct accagcacag ctcgacgagg aaagcccccc acaaaggagc ctatcagcca
     1741 gaccacggat gatatccttg tggcctcggc agagtgtccc agcgacgatg aggacattga
     1801 cccctgtgag ccgagctcag gtgggttagc caaccccacc cgagtgggcg gccgcgaacc
     1861 atacccaggc tcggcagagg tgatccggga gtctagcagt accactggca tggtggtggg
     1921 gattgtcgca gcagcagctc tgtgcatcct catcctcctc tatgccatgt acaagtacag
     1981 aaaccgggat gaaggctcat atcacgtgga tgagagtcga aactacatca gtaactcagc
     2041 acagtccaat ggggctgtgg tcaaggagaa gcagcccagc agtgcgaaaa gcgccaacaa
     2101 aaacaagaag aacaaggata aggagtatta tgtctgaatt cctgcagccc cgataatcaa
     2161 cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt
     2221 acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct
     2281 ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc
     2341 gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg
     2401 ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc
     2461 acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc
     2521 actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct gctcgcctgt
     2581 gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca
     2641 gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt
     2701 cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgggg ggatcctgtg
     2761 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     2821 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
     2881 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
     2941 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cggccgcagg aacccctagt
     3001 gatggagttg gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa
     3061 ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg
     3121 cctgcagggg cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     3181 catacgtcaa agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3241 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3301 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3361 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg
     3421 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3481 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     3541 tcgggctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     3601 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta
     3661 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     3721 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3781 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3841 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     3901 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3961 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4021 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4081 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     4141 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     4201 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     4261 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     4321 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     4381 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     4441 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     4501 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     4561 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     4621 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     4681 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     4741 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     4801 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     4861 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     4921 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     4981 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5041 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     5101 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     5161 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     5221 gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     5281 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     5341 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     5401 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     5461 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     5521 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     5581 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     5641 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     5701 ttttgctggc cttttgctca catgt
//
