LOCUS Exported 5714 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION An AAV vector that expresses yellow pre-eGRASP(p32) under the tetO promoter.. ACCESSION . VERSION . KEYWORDS pAAV-TIWB-yellow pre-eGRASP(p32) 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5714 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" 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 4034..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 //