LOCUS       sequence-212624-        5714 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 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(p30)
FEATURES             Location/Qualifiers
     source          1..5714
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   47..176
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        192..570
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     protein_bind    200..218
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    236..254
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    272..290
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    308..326
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    344..362
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    380..398
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    416..434
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     479..503
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          618..750
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     733..752
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        795..813
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      817..826
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             901..1359
                     /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(940..961)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     misc_feature    2190..2778
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2243..2263)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2661..2672)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2791..3015
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3023..3163
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3023..3152
                     /label=AAV2 ITR
     rep_origin      3238..3693
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3325..3344)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3535..3556
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3710..3729)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3829..3851
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3889..3907)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3975..4079
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4080..4148,4149..4940)
                     /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(4298..4317)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5111..5699
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5600..5619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtcctg caggcagctg
       61 cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg acctttggtc gcccggcctc
      121 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg
      181 ccgcacgcgt tggctttact ccctatcagt gatagagaac gtatgaagag tttactccct
      241 atcagtgata gagaacgtat gcagacttta ctccctatca gtgatagaga acgtataagg
      301 agtttactcc ctatcagtga tagagaacgt atgaccagtt tactccctat cagtgataga
      361 gaacgtatct acagtttact ccctatcagt gatagagaac gtatatccag tttactccct
      421 atcagtgata gagaacgtat aagctttagg cgtgtacggt gggcgcctat aaaagcagag
      481 ctcgtttagt gaaccgtcag atcgcctgga gcaattccac aacacttttg tcttatacca
      541 actttccgta ccacttccta ccctcgtaaa gctagtcgaa cttaagctgc agaagttggt
      601 cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga gaccaataga
      661 aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct attggtctta
      721 ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat tacagctctt
      781 aaggctagag tacttaatac gactcactat aggctagcca ccatggagac cgacaccctc
      841 ctgctatggg tgctgctgct ctgggtgccc ggcagcaccg gcgacgcacc ggtcggaggc
      901 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac
      961 gtaaacggcc acaagttcag cgtgaggggc gagggcgagg gcgatgccac catcggcaag
     1021 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg
     1081 accaccctga cctacggcgt gcagtgcttc gcccgctacc ccgaccacat gaagcggcac
     1141 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat ctccttcaag
     1201 gacgacggca agtacaagac ccgcgccgtg gtgaagttcg agggcgacac cctggtgaac
     1261 cgcatcgagc tgaagggcac cgacttcaag gaggacggca acatcctggg gcacaagctg
     1321 gagtacaact tcaacagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc
     1381 aaggccaact tcaccgtgcg ccacaacgtg gaggacggca gcgtgcagct cgccgaccac
     1441 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg
     1501 agctaccaga ccgtgctgag caaagacccc aacgagaaga ccggtggatc tggaggcagt
     1561 ggcggatcta gagctcctac aaagcctcca ccactgccac ctggaggagg ttccggaggc
     1621 ggaagcggta ccgaagtgcc ttcctctatg acaactgagt cgacagccac tgccatgcag
     1681 tccgagatgt ccacctcaat catggagacc accacaaccc tggctaccag cacagctcga
     1741 cgaggaaagc cccccacaaa ggagcctatc agccagacca cggatgatat ccttgtggcc
     1801 tcggcagagt gtcccagcga cgatgaggac attgacccct gtgagccgag ctcaggtggg
     1861 ttagccaacc ccacccgagt gggcggccgc gaaccatacc caggctcggc agaggtgatc
     1921 cgggagtcta gcagtaccac tggcatggtg gtggggattg tcgcagcagc agctctgtgc
     1981 atcctcatcc tcctctatgc catgtacaag tacagaaacc gggatgaagg ctcatatcac
     2041 gtggatgaga gtcgaaacta catcagtaac tcagcacagt ccaatggggc tgtggtcaag
     2101 gagaagcagc ccagcagtgc gaaaagcgcc aacaaaaaca agaagaacaa ggataaggag
     2161 tattatgtct gaattcctgc agccccgata atcaacctct ggattacaaa atttgtgaaa
     2221 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2281 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2341 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2401 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2461 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2521 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2581 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2641 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2701 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2761 tttgggccgc ctccccgcat cggggggatc ctgtgccttc tagttgccag ccatctgttg
     2821 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2881 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2941 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     3001 cggtgggctc tatggcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
     3061 cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc
     3121 cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat
     3181 tttctcctta cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg
     3241 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3301 cacttgccag cgccttagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3361 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3421 ctttacggca cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat
     3481 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3541 tcttgttcca aactggaaca acactcaact ctatctcggg ctattctttt gatttataag
     3601 ggattttgcc gatttcggtc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3661 cgaattttaa caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct
     3721 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     3781 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     3841 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     3901 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     3961 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4021 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4081 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4141 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4201 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4261 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     4321 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     4381 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     4441 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     4501 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     4561 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     4621 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     4681 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     4741 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     4801 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     4861 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     4921 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     4981 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5041 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5101 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5161 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5221 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     5281 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5341 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5401 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5461 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5521 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5581 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5641 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5701 acgccagcaa cgcg
//
