LOCUS       sequence-472885-        6519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses EGFP tagged Tau AP 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 AP
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      29..38
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(35..37,38..40,41..751)
                     /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(80..101)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(341..360)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     688..709
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      1926..1935
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1939..2073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1976..1995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2030..2049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2142..2499
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2142..2333
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2165..2185)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2350..2485
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2519..2536)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2519..2535)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2528..2550)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2748..3203
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2835..2854)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3045..3066
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3220..3239)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3339..3361
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3399..3417)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3485..3589
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3590..3658,3659..4450)
                     /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(3808..3827)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4621..5209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5110..5129
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5363..5380
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5497..5518
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5533..5538,5539..5556,5557..5563)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5571..5587
                     /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     5576..5598
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5595..5611
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5595..5611
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        5640..6019
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6020..6223
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6174..6194
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6220..6244
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        6454..6472
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6454..6471
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6485..6504
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 caactgcacc tcggttctat cgatggtcgc caccatggtg agcaagggcg aggagctgtt
       61 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      121 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      181 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
      241 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      301 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      361 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      421 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
      481 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
      541 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
      601 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcaaccagt ccgccctgag
      661 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
      721 gatcactctc ggcatggacg agctgtacaa gggatccgct gagccccgcc aggagttcga
      781 agtgatggaa gatcacgctg ggacgtacgg gttgggggac aggaaagatc aggggggcta
      841 caccatgcac caagaccaag agggtgacac ggacgctggc ctgaaagctg aagaagcagg
      901 cattggagac gcccccagcc tggaagacga agctgctggt cacgtgaccc aagctcgcat
      961 ggtcagtaaa agcaaagacg ggactggaag cgatgacaaa aaagccaagg gggctgatgg
     1021 taaaacgaag atcgccgcac cgcggggagc agcccctcca ggccagaagg gccaggccaa
     1081 cgccaccagg attccagcaa aagccccgcc cgctccaaag gcaccaccca gctctggtga
     1141 acctccaaaa tcaggggatc gcagcggcta cagcgccccc ggcgccccag gcgctcccgg
     1201 cagccgctcc cgcgccccgt cccttccagc cccacccacc cgggagccca agaaggtggc
     1261 agtggtccgt gctccaccca aggcgccgtc ttccgccaag agccgcctgc agacagcccc
     1321 cgtgcccatg ccagacctga agaatgtcaa gtccaagatc ggctccactg agaacctgaa
     1381 gcaccagccg ggaggcggga aggtgcagat aattaataag aagctggatc ttagcaacgt
     1441 ccagtccaag tgtggctcaa aggataatat caaacacgtc ccgggaggcg gcagtgtgca
     1501 aatagtctac aaaccagttg acctgagcaa ggtgacctcc aagtgtggct cattaggcaa
     1561 catccatcat aaaccaggag gtggccaggt ggaagtaaaa tctgagaagc ttgacttcaa
     1621 ggacagagtc cagtcgaaga ttgggtccct ggacaatatc acccacgtcc ctggcggagg
     1681 aaataaaaag attgaaaccc acaagctgac cttccgcgag aacgccaaag ccaagacaga
     1741 ccacggggcg gagatcgtgt acaaggcgcc agtggtgtct ggggacacgg ctccacggca
     1801 tctcagcaat gtctcctcca ccggcagcat cgacatggta gacgcgcccc agctcgccac
     1861 gctagctgac gaggtgtctg cctccctggc caagcagggt ttgtgagtcg acctgcagaa
     1921 gcttggccgc catggcccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1981 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2041 aaactcatca atgtatctta tcatgtctgg atcgggaatt aattcggcgc agcaccatgg
     2101 cctgaaataa cctctgaaag aggaacttgg ttaggtacct tctgaggcgg aaagaaccag
     2161 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     2221 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2281 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2341 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2401 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
     2461 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctgttaac agcttggcac
     2521 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     2581 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     2641 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     2701 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag
     2761 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     2821 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     2881 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     2941 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata
     3001 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3061 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc
     3121 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa
     3181 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc
     3241 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     3301 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     3361 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     3421 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     3481 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     3541 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     3601 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     3661 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     3721 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     3781 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     3841 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     3901 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     3961 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     4021 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     4081 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     4141 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     4201 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     4261 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     4321 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     4381 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     4441 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     4501 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     4561 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     4621 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     4681 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     4741 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     4801 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     4861 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     4921 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     4981 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5041 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     5101 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     5161 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     5221 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     5281 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     5341 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     5401 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     5461 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     5521 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     5581 ataacaattt cacacaggaa acagctatga catgattacg aattaattcg agctcgcccg
     5641 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     5701 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     5761 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     5821 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     5881 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     5941 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
     6001 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
     6061 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     6121 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
     6181 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     6241 gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc
     6301 cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt
     6361 aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac gcggctacaa
     6421 ttaatacata accttatgta tcatacacat acgatttagg tgacactata gaataacatc
     6481 cactttgcct ttctctccac aggtgtccac tcccaggtc
//
