LOCUS       sequence-242684-        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"
     protein_bind    85..106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(121..126,127..144,145..151)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    159..175
                     /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     164..186
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     183..199
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     183..199
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        228..607
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        608..811
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     762..782
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     808..832
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1042..1060
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1042..1059
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1073..1092
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1136..1145
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1142..1144,1145..1147,1148..1858)
                     /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(1187..1208)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1448..1467)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1795..1816
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3033..3042
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    3046..3180
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3083..3102)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3137..3156
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3249..3606
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3249..3440
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3272..3292)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3457..3592
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3626..3643)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3626..3642)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3635..3657)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3855..4310
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3942..3961)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4152..4173
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4327..4346)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4446..4468
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4506..4524)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4592..4696
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4697..4765,4766..5557)
                     /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(4915..4934)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5728..6316
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6217..6236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6470..6487
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
       61 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
      121 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
      181 cacaggaaac agctatgaca tgattacgaa ttaattcgag ctcgcccgac attgattatt
      241 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
      301 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
      361 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
      421 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
      481 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
      541 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
      601 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
      661 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
      721 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
      781 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcgcctggag
      841 acgccatcca cgctgttttg acctccatag aagacaccgg gaccgatcca gcctccgcgg
      901 ccgggaacgg tgcattggaa cgcggattcc ccgtgccaag agtgacgtaa gtaccgccta
      961 tagagtctat aggcccaccc ccttggcttc gttagaacgc ggctacaatt aatacataac
     1021 cttatgtatc atacacatac gatttaggtg acactataga ataacatcca ctttgccttt
     1081 ctctccacag gtgtccactc ccaggtccaa ctgcacctcg gttctatcga tggtcgccac
     1141 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1201 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1261 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1321 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     1381 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1441 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1501 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1561 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     1621 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1681 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1741 ctacctgagc aaccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1801 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaaggg
     1861 atccgctgag ccccgccagg agttcgaagt gatggaagat cacgctggga cgtacgggtt
     1921 gggggacagg aaagatcagg ggggctacac catgcaccaa gaccaagagg gtgacacgga
     1981 cgctggcctg aaagctgaag aagcaggcat tggagacgcc cccagcctgg aagacgaagc
     2041 tgctggtcac gtgacccaag ctcgcatggt cagtaaaagc aaagacggga ctggaagcga
     2101 tgacaaaaaa gccaaggggg ctgatggtaa aacgaagatc gccgcaccgc ggggagcagc
     2161 ccctccaggc cagaagggcc aggccaacgc caccaggatt ccagcaaaag ccccgcccgc
     2221 tccaaaggca ccacccagct ctggtgaacc tccaaaatca ggggatcgca gcggctacag
     2281 cgcccccggc gccccaggcg ctcccggcag ccgctcccgc gccccgtccc ttccagcccc
     2341 acccacccgg gagcccaaga aggtggcagt ggtccgtgct ccacccaagg cgccgtcttc
     2401 cgccaagagc cgcctgcaga cagcccccgt gcccatgcca gacctgaaga atgtcaagtc
     2461 caagatcggc tccactgaga acctgaagca ccagccggga ggcgggaagg tgcagataat
     2521 taataagaag ctggatctta gcaacgtcca gtccaagtgt ggctcaaagg ataatatcaa
     2581 acacgtcccg ggaggcggca gtgtgcaaat agtctacaaa ccagttgacc tgagcaaggt
     2641 gacctccaag tgtggctcat taggcaacat ccatcataaa ccaggaggtg gccaggtgga
     2701 agtaaaatct gagaagcttg acttcaagga cagagtccag tcgaagattg ggtccctgga
     2761 caatatcacc cacgtccctg gcggaggaaa taaaaagatt gaaacccaca agctgacctt
     2821 ccgcgagaac gccaaagcca agacagacca cggggcggag atcgtgtaca aggcgccagt
     2881 ggtgtctggg gacacggctc cacggcatct cagcaatgtc tcctccaccg gcagcatcga
     2941 catggtagac gcgccccagc tcgccacgct agctgacgag gtgtctgcct ccctggccaa
     3001 gcagggtttg tgagtcgacc tgcagaagct tggccgccat ggcccaactt gtttattgca
     3061 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     3121 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     3181 gggaattaat tcggcgcagc accatggcct gaaataacct ctgaaagagg aacttggtta
     3241 ggtaccttct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3301 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3361 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3421 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3541 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3601 tgcaaaaagc tgttaacagc ttggcactgg ccgtcgtttt acaacgtcgt gactgggaaa
     3661 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
     3721 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
     3781 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     3841 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     3901 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     3961 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4021 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4081 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4141 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta
     4201 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     4261 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4321 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4381 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4441 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4501 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4561 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4621 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4681 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4741 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4801 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     4861 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     4921 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     4981 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5041 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5101 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5161 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5221 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5281 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5341 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5401 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5461 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5521 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5581 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5641 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5701 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5761 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     5821 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     5881 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     5941 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6001 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6061 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6121 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6181 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6241 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6301 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6361 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     6421 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     6481 gagcgaggaa gcggaagagc gcccaatacg caaaccgcc
//
