LOCUS Exported 6519 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION expresses EGFP tagged Tau in mammalian cells. ACCESSION . VERSION . KEYWORDS pRK5-EGFP-Tau SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6519) AUTHORS Hoover BR, Reed MN, Su J, Penrod RD, Kotilinek LA, Grant MK, Pitstick R, Carlson GA, Lanier LM, Yuan LL, Ashe KH, Liao D TITLE Tau mislocalization to dendritic spines mediates synaptic dysfunction independently of neurodegeneration. JOURNAL Neuron. 2010 Dec 22;68(6):1067-81. doi: 10.1016/j.neuron.2010.11.030. PUBMED 21172610 REFERENCE 2 (bases 1 to 6519) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6519 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 94..111 /label=L4440 /note="L4440 vector, forward primer" protein_bind 228..249 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 264..294 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 302..318 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 307..329 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 326..342 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 326..342 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" enhancer 371..750 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 751..954 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 905..925 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 951..975 /label=LNCX /note="Human CMV promoter, forward primer" promoter 1185..1203 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 1185..1202 /label=SP6 /note="SP6 promoter, forward primer" primer_bind 1216..1235 /label=pMT2-F /note="Synthetic intron, forward primer" regulatory 1279..1288 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1285..2001 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSNQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1330..1351) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1591..1610) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1938..1959 /label=EGFP-C /note="EGFP, forward primer" regulatory 3176..3185 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 3189..3323 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3226..3245) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3280..3299 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" promoter 3392..3749 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(3415..3435) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 3600..3735 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3662..3681 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(3769..3786) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3769..3785) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3778..3800) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3998..4453 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4085..4104) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4295..4316 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4470..4489) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4589..4611 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4649..4667) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4735..4839 /gene="bla" /label=AmpR promoter CDS 4840..5700 /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(5058..5077) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5871..6459 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6360..6379 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct 61 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa 121 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 181 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 241 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 301 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattaattc 361 gagctcgccc gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta 421 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc 481 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg 541 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg 601 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa 661 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac 721 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg 781 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg 841 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca 901 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta 961 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac 1021 cgggaccgat ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc 1081 aagagtgacg taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa 1141 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat 1201 agaataacat ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc 1261 tcggttctat cgatggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg 1321 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc 1381 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc 1441 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc 1501 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc 1561 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag 1621 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag 1681 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat 1741 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc 1801 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc 1861 cccgtgctgc tgcccgacaa ccactacctg agcaaccagt ccgccctgag caaagacccc 1921 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc 1981 ggcatggacg agctgtacaa gggatccgct gagccccgcc aggagttcga agtgatggaa 2041 gatcacgctg ggacgtacgg gttgggggac aggaaagatc aggggggcta caccatgcac 2101 caagaccaag agggtgacac ggacgctggc ctgaaagctg aagaagcagg cattggagac 2161 acccccagcc tggaagacga agctgctggt cacgtgaccc aagctcgcat ggtcagtaaa 2221 agcaaagacg ggactggaag cgatgacaaa aaagccaagg gggctgatgg taaaacgaag 2281 atcgccacac cgcggggagc agcccctcca ggccagaagg gccaggccaa cgccaccagg 2341 attccagcaa aaaccccgcc cgctccaaag acaccaccca gctctggtga acctccaaaa 2401 tcaggggatc gcagcggcta cagcagcccc ggctccccag gcactcccgg cagccgctcc 2461 cgcaccccgt cccttccaac cccacccacc cgggagccca agaaggtggc agtggtccgt 2521 actccaccca agtcgccgtc ttccgccaag agccgcctgc agacagcccc cgtgcccatg 2581 ccagacctga agaatgtcaa gtccaagatc ggctccactg agaacctgaa gcaccagccg 2641 ggaggcggga aggtgcagat aattaataag aagctggatc ttagcaacgt ccagtccaag 2701 tgtggctcaa aggataatat caaacacgtc ccgggaggcg gcagtgtgca aatagtctac 2761 aaaccagttg acctgagcaa ggtgacctcc aagtgtggct cattaggcaa catccatcat 2821 aaaccaggag gtggccaggt ggaagtaaaa tctgagaagc ttgacttcaa ggacagagtc 2881 cagtcgaaga ttgggtccct ggacaatatc acccacgtcc ctggcggagg aaataaaaag 2941 attgaaaccc acaagctgac cttccgcgag aacgccaaag ccaagacaga ccacggggcg 3001 gagatcgtgt acaagtcgcc agtggtgtct ggggacacgt ctccacggca tctcagcaat 3061 gtctcctcca ccggcagcat cgacatggta gactcgcccc agctcgccac gctagctgac 3121 gaggtgtctg cctccctggc caagcagggt ttgtgagtcg acctgcagaa gcttggccgc 3181 catggcccaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 3241 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 3301 atgtatctta tcatgtctgg atcgggaatt aattcggcgc agcaccatgg cctgaaataa 3361 cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag ctgtggaatg 3421 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca 3481 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa 3541 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca 3601 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt 3661 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag 3721 gcttttttgg aggcctaggc ttttgcaaaa agctgttaac agcttggcac tggccgtcgt 3781 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca 3841 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca 3901 gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta cgcatctgtg 3961 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta 4021 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg 4081 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa 4141 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc 4201 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt 4261 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca 4321 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc 4381 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta 4441 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc 4501 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca 4561 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg 4621 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat 4681 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga 4741 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa 4801 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt 4861 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg 4921 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg 4981 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg 5041 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag 5101 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca 5161 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg 5221 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc 5281 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg 5341 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg 5401 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac 5461 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg 5521 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg 5581 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact 5641 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa 5701 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt 5761 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag 5821 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct 5881 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt 5941 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg 6001 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct 6061 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc 6121 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg 6181 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa 6241 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg 6301 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg 6361 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga 6421 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt 6481 ttacggttcc tggccttttg ctggcctttt gctcacatg //