pRK5-EGFP-Tau P301L

LOCUS       pRK5-EGFP-Tau P301L                6519 bp ds-DNA     circular SYN 06-AUG-2018
DEFINITION  expresses EGFP tagged Tau P301L in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pRK5-EGFP-Tau P301L
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 Aug 6, 2018 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6519
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     830..854
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1064..1082
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1064..1081
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1095..1114
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1158..1167
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1164..1880
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1209..1230)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1470..1489)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1817..1838
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      3055..3064
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     polyA_signal    3068..3202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3105..3124)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3159..3178
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        3271..3628
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3294..3314)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3479..3614
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3541..3560
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3648..3665)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3648..3664)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3657..3679)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3877..4332
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3964..3983)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4174..4195
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4349..4368)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4468..4490
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4528..4546)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4614..4718
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4719..5579
                     /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(4937..4956)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5750..6338
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6239..6258
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6492..6509
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
      241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
      961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
     1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
     1141 cggttctatc gatggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     1201 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     1261 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     1321 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     1381 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     1441 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     1501 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     1561 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     1621 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     1681 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     1741 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     1801 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     1861 gcatggacga gctgtacaag ggatccgctg agccccgcca ggagttcgaa gtgatggaag
     1921 atcacgctgg gacgtacggg ttgggggaca ggaaagatca ggggggctac accatgcacc
     1981 aagaccaaga gggtgacacg gacgctggcc tgaaagctga agaagcaggc attggagaca
     2041 cccccagcct ggaagacgaa gctgctggtc acgtgaccca agctcgcatg gtcagtaaaa
     2101 gcaaagacgg gactggaagc gatgacaaaa aagccaaggg ggctgatggt aaaacgaaga
     2161 tcgccacacc gcggggagca gcccctccag gccagaaggg ccaggccaac gccaccagga
     2221 ttccagcaaa aaccccgccc gctccaaaga caccacccag ctctggtgaa cctccaaaat
     2281 caggggatcg cagcggctac agcagccccg gctccccagg cactcccggc agccgctccc
     2341 gcaccccgtc ccttccaacc ccacccaccc gggagcccaa gaaggtggca gtggtccgta
     2401 ctccacccaa gtcgccgtct tccgccaaga gccgcctgca gacagccccc gtgcccatgc
     2461 cagacctgaa gaatgtcaag tccaagatcg gctccactga gaacctgaag caccagccgg
     2521 gaggcgggaa ggtgcagata attaataaga agctggatct tagcaacgtc cagtccaagt
     2581 gtggctcaaa ggataatatc aaacacgtcc tgggaggcgg cagtgtgcaa atagtctaca
     2641 aaccagttga cctgagcaag gtgacctcca agtgtggctc attaggcaac atccatcata
     2701 aaccaggagg tggccaggtg gaagtaaaat ctgagaagct tgacttcaag gacagagtcc
     2761 agtcgaagat tgggtccctg gacaatatca cccacgtccc tggcggagga aataaaaaga
     2821 ttgaaaccca caagctgacc ttccgcgaga acgccaaagc caagacagac cacggggcgg
     2881 agatcgtgta caagtcgcca gtggtgtctg gggacacgtc tccacggcat ctcagcaatg
     2941 tctcctccac cggcagcatc gacatggtag actcgcccca gctcgccacg ctagctgacg
     3001 aggtgtctgc ctccctggcc aagcagggtt tgtgagtcga cctgcagaag cttggccgcc
     3061 atggcccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     3121 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     3181 tgtatcttat catgtctgga tcgggaatta attcggcgca gcaccatggc ctgaaataac
     3241 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     3301 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3361 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     3421 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     3481 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     3541 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     3601 cttttttgga ggcctaggct tttgcaaaaa gctgttaaca gcttggcact ggccgtcgtt
     3661 ttacaacgtc gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat
     3721 ccccctttcg ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag
     3781 ttgcgcagcc tgaatggcga atggcgcctg atgcggtatt ttctccttac gcatctgtgc
     3841 ggtatttcac accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa
     3901 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     3961 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     4021 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     4081 aaaaacttga tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     4141 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     4201 cactcaaccc tatctcgggc tattcttttg atttataagg gattttgccg atttcggcct
     4261 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     4321 cgtttacaat tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     4381 agccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     4441 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     4501 catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     4561 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     4621 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     4681 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     4741 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     4801 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     4861 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     4921 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     4981 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5041 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5101 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5161 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5221 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5281 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5341 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     5401 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     5461 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     5521 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     5581 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5641 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5701 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5761 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5821 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5881 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     5941 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6001 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6061 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6121 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6181 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6241 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6301 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6361 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6421 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     6481 cgaccgagcg cagcgagtca gtgagcgagg aagcggaag
//