LOCUS       sequence-450730-        8226 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses tau propagation ORF with no V5 tag on WT 2N4R tau.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8226)
  AUTHORS   Dickson JR, Sobolewski RGR, Fernandes AR, Cooper JM, Fan Z, Chung M,
            Donahue C, Oakley DH, Strickland DK, Hyman BT
  TITLE     Alzheimer disease-associated tau post-translational modification 
            mimics impact tau propagation and uptake.
  JOURNAL   J Neuropathol Exp Neurol. 2025 Jun 1;84(6):459-470. doi: 
            10.1093/jnen/nlaf007.
  PUBMED    39984820
REFERENCE   2  (bases 1 to 8226)
  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"; journalName: 
            "J Neuropathol Exp Neurol"; date: "2025-06-1- 1"; volume: "84"; 
            issue: "6"; pages: "459-470"
COMMENT     Sequence Label: pcDNA3.1(-).TagRFP.T-2A-Tau2N4R-WPRE
FEATURES             Location/Qualifiers
     source          1..8226
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    921..973
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      981..990
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             987..1718
                     /codon_start=1
                     /product="photostable variant of TagRFP (Shaner et al., 
                     2008)"
                     /label=TagRFP-T
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRTD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     CDS             1728..1781
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     misc_feature    3136..3724
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3189..3209)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3607..3618)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3817..3834)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3823..4047
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4093..4521
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4180..4199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4390..4411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4530..4550)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4535..4864
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4715..4850
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4777..4796
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4931..5725
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4985..5004)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5595..5614
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5899..6020
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5936..5955)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5990..6009
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6069..6085)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6069..6085)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6082..6104)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6093..6109
                     /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(6117..6123,6124..6141,6142..6147))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6162..6183
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6300..6317)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6471..7059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6551..6570)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7230..8021,8022..8090))
                     /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     7853..7872
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8091..8195)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagctca agcttcgaat tctgcagtcg acggtaccgc
      961 gggcccggga tccaccggtc gccaccatgg tgtctaaggg cgaagagctg attaaggaga
     1021 acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc aagtgcacat
     1081 ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg
     1141 gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac ggcagcagaa
     1201 ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc cctgagggct
     1261 tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca
     1321 ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcccat
     1381 ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggccaacacc gagatgctgt
     1441 accccgctga cggcggcctg gaaggcagaa ccgacatggc cctgaagctc gtgggcgggg
     1501 gccacctgat ctgcaacttc aagaccacat acagatccaa gaaacccgct aagaacctca
     1561 agatgcccgg cgtctactat gtggaccaca gactggaaag aatcaaggag gccgacaaag
     1621 agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc cctagcaaac
     1681 tggggcacaa acttaatggc atggacgagc tgtacaaggg taccggcgag ggcagaggaa
     1741 gtctgctaac atgcggtgac gtcgaggaga atcctggccc aaccggtaaa actagtgctg
     1801 agccccgcca ggagttcgaa gtgatggaag atcacgctgg gacgtacggg ttgggggaca
     1861 ggaaagatca ggggggctac accatgcacc aagaccaaga gggtgacacg gacgctggcc
     1921 tgaaagaatc tcccctgcag acccccactg aggacggatc tgaggaaccg ggctctgaaa
     1981 cctctgatgc taagagcact ccaacagcgg aagatgtgac agcaccctta gtggatgagg
     2041 gagctcccgg caagcaggct gccgcgcagc cccacacgga gatcccagaa ggaaccacag
     2101 ctgaagaagc aggcattgga gacaccccca gcctggaaga cgaagctgct ggtcacgtga
     2161 cccaagctcg catggtcagt aaaagcaaag acgggactgg aagcgatgac aaaaaagcca
     2221 agggggctga tggtaaaacg aagatcgcca caccgcgggg agcagcccct ccaggccaga
     2281 agggccaggc caacgccacc aggattccag caaaaacccc gcccgctcca aagacaccac
     2341 ccagctctgg tgaacctcca aaatcagggg atcgcagcgg ctacagcagc cccggctccc
     2401 caggcactcc cggcagccgc tcccgcaccc cgtcccttcc aaccccaccc acccgggagc
     2461 ccaagaaggt ggcagtggtc cgtactccac ccaagtcgcc gtcttccgcc aagagccgcc
     2521 tgcagacagc ccccgtgccc atgccagacc tgaagaatgt caagtccaag atcggctcca
     2581 ctgagaacct gaagcaccag ccgggaggcg ggaaggtgca gataattaat aagaagctgg
     2641 atcttagcaa cgtccagtcc aagtgtggct caaaggataa tatcaaacac gtcccgggag
     2701 gcggcagtgt gcaaatagtc tacaaaccag ttgacctgag caaggtgacc tccaagtgtg
     2761 gctcattagg caacatccat cataaaccag gaggtggcca ggtggaagta aaatctgaga
     2821 agcttgactt caaggacaga gtccagtcga agattgggtc cctggacaat atcacccacg
     2881 tccctggcgg aggaaataaa aagattgaaa cccacaagct gaccttccgc gagaacgcca
     2941 aagccaagac agaccacggg gcggagatcg tgtacaagtc gccagtggtg tctggggaca
     3001 cgtctccacg gcatctcagc aatgtctcct ccaccggcag catcgacatg gtagactcgc
     3061 cccagctcgc cacgctagct gacgaggtgt ctgcctccct ggccaagcag ggtttgtagg
     3121 gatccgatta tcgataatca acctctggat tacaaaattt gtgaaagatt gactggtatt
     3181 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat
     3241 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct
     3301 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct
     3361 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc
     3421 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     3481 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa atcatcgtcc
     3541 tttccttggc tgctcgcctg tgttgccacc tggattctgc gcgggacgcc cttctgctac
     3601 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg
     3661 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc
     3721 ccgcatcgat accgtcgacg atatctgcag aattccacca cactggacta gtggatccga
     3781 gctcggtacc aagcttaagt ttaaaccgct gatcagcctc gactgtgcct tctagttgcc
     3841 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     3901 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     3961 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     4021 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     4081 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4141 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4201 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4261 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     4321 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4381 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     4441 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4501 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4561 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4621 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4681 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4741 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4801 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4861 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     4921 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     4981 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     5041 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     5101 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     5161 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     5221 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     5281 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     5341 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     5401 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     5461 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     5521 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     5581 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     5641 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     5701 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     5761 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     5821 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     5881 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5941 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     6001 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     6061 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     6121 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     6181 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     6241 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     6301 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     6361 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     6421 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6481 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6541 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6601 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6661 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6721 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6781 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6841 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6901 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6961 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     7021 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     7081 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     7141 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     7201 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     7261 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     7321 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     7381 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     7441 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     7501 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     7561 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     7621 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     7681 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     7741 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     7801 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     7861 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     7921 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     7981 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     8041 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     8101 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     8161 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     8221 gacgtc
//
