LOCUS       sequence-212642-        5714 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  An AAV vector that expresses yellow pre-eGRASP(p32) under the tetO 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5714)
  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 5714)
  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(p32)
FEATURES             Location/Qualifiers
     source          1..5714
                     /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        146..524
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    154..172
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    190..208
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    262..280
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    298..316
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    334..352
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    370..388
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     433..457
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          572..704
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     687..706
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        749..767
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      771..780
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             855..1313
                     /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(894..915)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2144..2732
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2197..2217)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2615..2626)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2745..2969
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2977..3117
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2977..3106
                     /label=AAV2 ITR
     rep_origin      3192..3647
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3279..3298)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3489..3510
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3664..3683)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3783..3805
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3843..3861)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3929..4033
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4034..4102,4103..4894)
                     /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(4252..4271)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5065..5653
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5554..5573
                     /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 gcgttggctt tactccctat cagtgataga gaacgtatga
      181 agagtttact ccctatcagt gatagagaac gtatgcagac tttactccct atcagtgata
      241 gagaacgtat aaggagttta ctccctatca gtgatagaga acgtatgacc agtttactcc
      301 ctatcagtga tagagaacgt atctacagtt tactccctat cagtgataga gaacgtatat
      361 ccagtttact ccctatcagt gatagagaac gtataagctt taggcgtgta cggtgggcgc
      421 ctataaaagc agagctcgtt tagtgaaccg tcagatcgcc tggagcaatt ccacaacact
      481 tttgtcttat accaactttc cgtaccactt cctaccctcg taaagctagt cgaacttaag
      541 ctgcagaagt tggtcgtgag gcactgggca ggtaagtatc aaggttacaa gacaggttta
      601 aggagaccaa tagaaactgg gcttgtcgag acagagaaga ctcttgcgtt tctgataggc
      661 acctattggt cttactgaca tccactttgc ctttctctcc acaggtgtcc actcccagtt
      721 caattacagc tcttaaggct agagtactta atacgactca ctataggcta gccaccatgg
      781 agaccgacac cctcctgcta tgggtgctgc tgctctgggt gcccggcagc accggcgacg
      841 caccggtcgg aggcagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      901 agctggacgg cgacgtaaac ggccacaagt tcagcgtgag gggcgagggc gagggcgatg
      961 ccaccatcgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     1021 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcgcccgc taccccgacc
     1081 acatgaagcg gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     1141 ccatctcctt caaggacgac ggcaagtaca agacccgcgc cgtggtgaag ttcgagggcg
     1201 acaccctggt gaaccgcatc gagctgaagg gcaccgactt caaggaggac ggcaacatcc
     1261 tggggcacaa gctggagtac aacttcaaca gccacaacgt ctatatcacc gccgacaagc
     1321 agaagaacgg catcaaggcc aacttcaccg tgcgccacaa cgtggaggac ggcagcgtgc
     1381 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     1441 acaaccacta cctgagctac cagaccgtgc tgagcaaaga ccccaacgag aagaccggtg
     1501 gatctggagg cagtggcgga tctagatctc ctagctactc tccaccacct ccacctggag
     1561 gaggttccgg aggcggaagc ggtaccgaag tgccttcctc tatgacaact gagtcgacag
     1621 ccactgccat gcagtccgag atgtccacct caatcatgga gaccaccaca accctggcta
     1681 ccagcacagc tcgacgagga aagcccccca caaaggagcc tatcagccag accacggatg
     1741 atatccttgt ggcctcggca gagtgtccca gcgacgatga ggacattgac ccctgtgagc
     1801 cgagctcagg tgggttagcc aaccccaccc gagtgggcgg ccgcgaacca tacccaggct
     1861 cggcagaggt gatccgggag tctagcagta ccactggcat ggtggtgggg attgtcgcag
     1921 cagcagctct gtgcatcctc atcctcctct atgccatgta caagtacaga aaccgggatg
     1981 aaggctcata tcacgtggat gagagtcgaa actacatcag taactcagca cagtccaatg
     2041 gggctgtggt caaggagaag cagcccagca gtgcgaaaag cgccaacaaa aacaagaaga
     2101 acaaggataa ggagtattat gtctgaattc ctgcagcccc gataatcaac ctctggatta
     2161 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2221 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2281 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2341 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2401 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2461 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2521 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     2581 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2641 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     2701 gagtcggatc tccctttggg ccgcctcccc gcatcggggg gatcctgtgc cttctagttg
     2761 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     2821 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     2881 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     2941 gcatgctggg gatgcggtgg gctctatggc ggccgcagga acccctagtg atggagttgg
     3001 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     3061 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     3121 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     3181 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3241 cagcgtgacc gctacacttg ccagcgcctt agcgcccgct cctttcgctt tcttcccttc
     3301 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3361 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     3421 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3481 ctttaatagt ggactcttgt tccaaactgg aacaacactc aactctatct cgggctattc
     3541 ttttgattta taagggattt tgccgatttc ggtctattgg ttaaaaaatg agctgattta
     3601 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     3661 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     3721 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     3781 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     3841 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3901 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3961 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     4021 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     4081 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     4141 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     4201 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     4261 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     4321 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4381 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4441 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4501 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4561 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4621 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4681 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4741 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4801 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4861 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4921 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4981 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5041 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5101 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5161 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5221 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5281 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5341 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5401 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5461 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5521 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5581 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5641 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5701 ttttgctcac atgt
//
