LOCUS       Exported                6519 bp ds-DNA     circular SYN 13-MAY-2021
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
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"
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"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1164..1880
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1164..1880
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /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"
                     /label=Kozak sequence
                     /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"