LOCUS       sequence-364531-        6519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses EGFP tagged Tau E14 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 E14
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     279..298
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     538..559
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(654..673)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(686..704)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(687..704)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(914..938)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(935..1138)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(964..984)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        1139..1518
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     complement(1547..1563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1547..1563)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1560..1582)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1571..1587
                     /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(1595..1601,1602..1619,1620..1625))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1640..1661
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1778..1795)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1949..2537)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2029..2048)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2708..3499,3500..3568))
                     /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     3331..3350
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3569..3673)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3741..3759
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3797..3819)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3919..3938
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(3955..4410)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4092..4113)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4304..4323
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4608..4630
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4622..4639
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4623..4639
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(4659..5016)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4673..4808
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4825..5016
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4973..4993
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    5085..5219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5109..5128)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5163..5182
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     regulatory      5223..5232
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(join(6407..6519,1..598,599..601,602..604))
                     /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(6449..6470)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc tgctggtagt
       61 ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga
      121 tgccgttctt ctgcttgtcg gccatgatat agacgttgtg gctgttgtag ttgtactcca
      181 gcttgtgccc caggatgttg ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt
      241 tcaccagggt gtcgccctcg aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct
      301 tgaagaagat ggtgcgctcc tggacgtagc cttcgggcat ggcggacttg aagaagtcgt
      361 gctgcttcat gtggtcgggg tagcggctga agcactgcac gccgtaggtc agggtggtca
      421 cgagggtggg ccagggcacg ggcagcttgc cggtggtgca gatgaacttc agggtcagct
      481 tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt
      541 cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg cccttgctca
      601 ccatggtggc gaccatcgat agaaccgagg tgcagttgga cctgggagtg gacacctgtg
      661 gagagaaagg caaagtggat gttattctat agtgtcacct aaatcgtatg tgtatgatac
      721 ataaggttat gtattaattg tagccgcgtt ctaacgaagc caagggggtg ggcctataga
      781 ctctataggc ggtacttacg tcactcttgg cacggggaat ccgcgttcca atgcaccgtt
      841 cccggccgcg gaggctggat cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat
      901 ggcgtctcca ggcgatctga cggttcacta aacgagctct gcttatatag acctcccacc
      961 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     1021 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     1081 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     1141 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     1201 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     1261 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     1321 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     1381 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     1441 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     1501 tagtcaataa tcaatgtcgg gcgagctcga attaattcgt aatcatgtca tagctgtttc
     1561 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     1621 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     1681 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     1741 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     1801 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     1861 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     1921 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     1981 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2041 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2101 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2161 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     2221 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     2281 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     2341 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     2401 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     2461 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     2521 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     2581 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     2641 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     2701 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2761 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2821 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2881 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2941 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3001 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3061 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3121 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3181 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     3241 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     3301 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     3361 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     3421 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     3481 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     3541 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     3601 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     3661 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     3721 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg
     3781 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     3841 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     3901 ggggctggct taactatgcg gcatcagagc agattgtact gagagtgcac cataaaattg
     3961 taaacgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     4021 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagccc gagatagggt
     4081 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     4141 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca cccaaatcaa
     4201 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     4261 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     4321 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     4381 ccgcgcttaa tgcgccgcta cagggcgcgt actatggttg ctttgacgta tgcggtgtga
     4441 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     4501 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     4561 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     4621 ttgtaaaacg acggccagtg ccaagctgtt aacagctttt tgcaaaagcc taggcctcca
     4681 aaaaagcctc ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca
     4741 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg
     4801 ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt
     4861 tgcatacttc tgcctgctgg agagcctggg gactttccac acctggttgc tgactaattg
     4921 agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac
     4981 tgacacacat tccacagctg gttctttccg cctcagaagg tacctaacca agttcctctt
     5041 tcagaggtta tttcaggcca tggtgctgcg ccgaattaat tcccgatcca gacatgataa
     5101 gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt
     5161 gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagttg
     5221 ggccatggcg gccaagcttc tgcaggtcga ctcacaaacc ctgcttggcc agggaggcag
     5281 acacctcgtc agctagcgtg gcgagctggg gctcgtctac catgtcgatg ctgccggtgg
     5341 aggagacatt gctgagatgc cgtggttccg tgtccccaga caccactggc tccttgtaca
     5401 cgatctccgc cccgtggtct gtcttggctt tggcgttctc gcggaaggtc agcttgtggg
     5461 tttcaatctt tttatttcct ccgccaggga cgtgggtgat attgtccagg gacccaatct
     5521 tcgactggac tctgtccttg aagtcaagct tctcagattt tacttccacc tggccacctc
     5581 ctggtttatg atggatgttg cctaatgagc cacacttgga ggtcaccttg ctcaggtcaa
     5641 ctggtttgta gactatttgc acactgccgc ctcccgggac gtgtttgata ttatcctttg
     5701 agccacactt ggactggacg ttgctaagat ccagcttctt attaattatc tgcaccttcc
     5761 cgcctcccgg ctggtgcttc aggttctcag tggagccgat cttggacttg acattcttca
     5821 ggtctggcat gggcacgggg gctgtctgca ggcggctctt ggcggaagac ggctccttgg
     5881 gtggctcacg gaccactgcc accttcttgg gctcccgggt gggtggttct ggaagggacg
     5941 gttcgcggga gcggctgccg ggttcgcctg gttcgccggg ttcgctgtag ccgctgcgat
     6001 cccctgattt tggaggttca ccagagctgg gtggttcctt tggagcgggc ggttcttttg
     6061 ctggaatcct ggtggcgttg gcctggccct tctggcctgg aggggctgct ccccgcggtt
     6121 cggcgatctt cgttttacca tcagccccct tggctttttt gtcatcgctt ccagtcccgt
     6181 ctttgctttt actgaccatg cgagcttggg tcacgtgacc agcagcttcg tcttccaggc
     6241 tgggttcgtc tccaatgcct gcttcttcag ctttcaggcc agcgtccgtg tcaccctctt
     6301 ggtcttggtg catggtgtag cccccctgat ctttcctgtc ccccaacccg tacgtcccag
     6361 cgtgatcttc catcacttcg aactcctggc ggggctcagc ggatcccttg tacagctcgt
     6421 ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct
     6481 tctcgttggg gtctttgctc agggcggact ggttgctca
//
