LOCUS       sequence-372929-        6519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses EGFP tagged Tau P301L in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2010.11"; journalName: "Neuron"; date: 
            "2010-12-22- 22"; volume: "68"; issue: "6"; pages: "1067-81"
COMMENT     Sequence Label: pRK5-EGFP-Tau P301L
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     69..89
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    181..315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(205..224)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     259..278
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      319..328
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(join(1503..2213,2214..2216,2217..2219))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1503..2213,2214..2216,2217..2219))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1545..1566)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1894..1913
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2153..2174
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2216..2225
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2269..2288)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2301..2319)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2302..2319)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2529..2553)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2550..2753)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2579..2599)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2754..3133
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(3162..3178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3162..3178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3175..3197)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3186..3202
                     /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)."
     promoter        complement(join(3210..3216,3217..3234,3235..3240))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3255..3276
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3393..3410)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3564..4152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3644..3663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4323..5114,5115..5183))
                     /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     4946..4965
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5184..5288)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5356..5374
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5412..5434)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5534..5553
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(5570..6025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5707..5728)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5919..5938
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6223..6245
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6237..6254
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6238..6254
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(join(6274..6519,1..112))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6288..6423
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        join(6440..6519,1..112)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
ORIGIN
        1 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
       61 ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc agaaggtacc
      121 taaccaagtt cctctttcag aggttatttc aggccatggt gctgcgccga attaattccc
      181 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      241 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
      301 tgcaataaac aagttgggcc atggcggcca agcttctgca ggtcgactca caaaccctgc
      361 ttggccaggg aggcagacac ctcgtcagct agcgtggcga gctggggcga gtctaccatg
      421 tcgatgctgc cggtggagga gacattgctg agatgccgtg gagacgtgtc cccagacacc
      481 actggcgact tgtacacgat ctccgccccg tggtctgtct tggctttggc gttctcgcgg
      541 aaggtcagct tgtgggtttc aatcttttta tttcctccgc cagggacgtg ggtgatattg
      601 tccagggacc caatcttcga ctggactctg tccttgaagt caagcttctc agattttact
      661 tccacctggc cacctcctgg tttatgatgg atgttgccta atgagccaca cttggaggtc
      721 accttgctca ggtcaactgg tttgtagact atttgcacac tgccgcctcc caggacgtgt
      781 ttgatattat cctttgagcc acacttggac tggacgttgc taagatccag cttcttatta
      841 attatctgca ccttcccgcc tcccggctgg tgcttcaggt tctcagtgga gccgatcttg
      901 gacttgacat tcttcaggtc tggcatgggc acgggggctg tctgcaggcg gctcttggcg
      961 gaagacggcg acttgggtgg agtacggacc actgccacct tcttgggctc ccgggtgggt
     1021 ggggttggaa gggacggggt gcgggagcgg ctgccgggag tgcctgggga gccggggctg
     1081 ctgtagccgc tgcgatcccc tgattttgga ggttcaccag agctgggtgg tgtctttgga
     1141 gcgggcgggg tttttgctgg aatcctggtg gcgttggcct ggcccttctg gcctggaggg
     1201 gctgctcccc gcggtgtggc gatcttcgtt ttaccatcag cccccttggc ttttttgtca
     1261 tcgcttccag tcccgtcttt gcttttactg accatgcgag cttgggtcac gtgaccagca
     1321 gcttcgtctt ccaggctggg ggtgtctcca atgcctgctt cttcagcttt caggccagcg
     1381 tccgtgtcac cctcttggtc ttggtgcatg gtgtagcccc cctgatcttt cctgtccccc
     1441 aacccgtacg tcccagcgtg atcttccatc acttcgaact cctggcgggg ctcagcggat
     1501 cccttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     1561 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag
     1621 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     1681 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     1741 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     1801 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     1861 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     1921 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     1981 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     2041 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     2101 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     2161 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     2221 gtggcgacca tcgatagaac cgaggtgcag ttggacctgg gagtggacac ctgtggagag
     2281 aaaggcaaag tggatgttat tctatagtgt cacctaaatc gtatgtgtat gatacataag
     2341 gttatgtatt aattgtagcc gcgttctaac gaagccaagg gggtgggcct atagactcta
     2401 taggcggtac ttacgtcact cttggcacgg ggaatccgcg ttccaatgca ccgttcccgg
     2461 ccgcggaggc tggatcggtc ccggtgtctt ctatggaggt caaaacagcg tggatggcgt
     2521 ctccaggcga tctgacggtt cactaaacga gctctgctta tatagacctc ccaccgtaca
     2581 cgcctaccgc ccatttgcgt caatggggcg gagttgttac gacattttgg aaagtcccgt
     2641 tgattttggt gccaaaacaa actcccattg acgtcaatgg ggtggagact tggaaatccc
     2701 cgtgagtcaa accgctatcc acgcccattg atgtactgcc aaaaccgcat caccatggta
     2761 atagcgatga ctaatacgta gatgtactgc caagtaggaa agtcccataa ggtcatgtac
     2821 tgggcataat gccaggcggg ccatttaccg tcattgacgt caataggggg cgtacttggc
     2881 atatgataca cttgatgtac tgccaagtgg gcagtttacc gtaaatactc cacccattga
     2941 cgtcaatgga aagtccctat tggcgttact atgggaacat acgtcattat tgacgtcaat
     3001 gggcgggggt cgttgggcgg tcagccaggc gggccattta ccgtaagtta tgtaacgcgg
     3061 aactccatat atgggctatg aactaatgac cccgtaattg attactatta ataactagtc
     3121 aataatcaat gtcgggcgag ctcgaattaa ttcgtaatca tgtcatagct gtttcctgtg
     3181 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     3241 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     3301 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     3361 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     3421 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3481 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3541 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3601 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3661 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3721 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3781 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3841 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3901 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     3961 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4021 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4081 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4141 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4201 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4261 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4321 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     4381 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     4441 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     4501 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     4561 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     4621 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     4681 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     4741 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     4801 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     4861 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     4921 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     4981 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5041 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5101 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5161 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5221 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5281 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     5341 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     5401 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5461 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5521 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccataa aattgtaaac
     5581 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     5641 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agcccgagat agggttgagt
     5701 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     5761 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccaa atcaagtttt
     5821 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5881 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5941 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     6001 cttaatgcgc cgctacaggg cgcgtactat ggttgctttg acgtatgcgg tgtgaaatac
     6061 cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca
     6121 actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg
     6181 gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta
     6241 aaacgacggc cagtgccaag ctgttaacag ctttttgcaa aagcctaggc ctccaaaaaa
     6301 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
     6361 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
     6421 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat
     6481 acttctgcct gctggggagc ctggggactt tccacacct
//
