LOCUS       sequence-212099-        6519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses EGFP tagged Tau 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
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        join(264..269,270..287,288..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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1285..1287,1288..1290,1291..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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     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"
     enhancer        3392..3583
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     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     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             join(4840..4908,4909..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
//
