LOCUS       sequence-267979-        8201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Lentiviral vector with EF1A promoter constitutively expressing a 
            single insert containing the Tau.K18(P301L/V337M)-mRuby2 fusion 
            protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8201)
  AUTHORS   Chen JJ, Nathaniel DL, Raghavan P, Nelson M, Tian R, Tse E, Hong JY,
            See SK, Mok SA, Hein MY, Southworth DR, Grinberg LT, Gestwicki JE, 
            Leonetti MD, Kampmann M
  TITLE     Compromised function of the ESCRT pathway promotes endolysosomal 
            escape of tau seeds and propagation of tau aggregation.
  JOURNAL   J Biol Chem. 2019 Oct 2. pii: RA119.009432. doi: 
            10.1074/jbc.RA119.009432.
  PUBMED    31578281
REFERENCE   2  (bases 1 to 8201)
  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"; journalName: "J Biol 
            Chem. 2019 Oct 2. pii: RA119.009432. doi: 10.1074/jbc.RA119.009432."
COMMENT     Sequence Label: pMK1254
FEATURES             Location/Qualifiers
     source          1..8201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(47..66)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        238..617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     772..792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     LTR             835..1015
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1684..1917
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2102..2146
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2444..2561
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2664..2875
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     intron          2876..3814
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     3771..3791
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             4290..4997
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTHM
                     ALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAVA
                     KFAGLGGGMDELYK"
     misc_feature    5090..5678
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5143..5163)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5561..5572)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5681..5697)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5682..5698)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             6207..6387
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(6449..7037)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6529..6548)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7208..7999,8000..8068))
                     /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     7831..7850
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8069..8173)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
      841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
      901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
      961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
     1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
     1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
     1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
     1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
     1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
     1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
     1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
     1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
     1501 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga
     1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
     1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
     1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
     1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
     1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
     1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
     1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
     1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
     2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
     2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
     2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
     2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
     2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
     2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
     2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
     2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
     2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
     2581 gcagagatcc agtttggtta gtaccgggcc cgctctaggg ccgcgtggat aaccgtatta
     2641 ccgccatgca tgtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
     2701 ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg taaactggga
     2761 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
     2821 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggtaag
     2881 tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt gcgtgccttg
     2941 aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg
     3001 gtgggagagt tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc
     3061 ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc
     3121 tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt
     3181 ctggcaagat agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg
     3241 ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct
     3301 gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt
     3361 gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc
     3421 accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg
     3481 gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt
     3541 tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct
     3601 cgattagttc tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc
     3661 gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat
     3721 gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca
     3781 gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgagctagcc accatgctgc
     3841 agacagcccc cgtgcccatg ccagacctga agaatgtcaa gtccaagatc ggctccactg
     3901 agaacctgaa gcaccagccg ggaggcggga aggtgcagat aattaataag aagctggatc
     3961 ttagcaacgt ccagtccaag tgtggctcaa aggataatat caaacacgtc ctgggaggcg
     4021 gcagtgtgca aatagtctac aaaccagttg acctgagcaa ggtgacctcc aagtgtggct
     4081 cattaggcaa catccatcat aaaccaggag gtggccagat ggaagtaaaa tctgagaagc
     4141 ttgacttcaa ggacagagtc cagtcgaaga ttgggtccct ggacaatatc acccacgtcc
     4201 ctggcggagg aaataaaaag attgaaaccc acaaggaatt ctgcagtcga cggtaccgcg
     4261 ggcccgggat ccaccggtcg ccaaccgcag tgtctaaggg cgaagagctg atcaaggaaa
     4321 atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc aaatgcacag
     4381 gtgaaggaga aggcaatccg tacatgggaa ctcaaaccat gaggatcaaa gtcatcgagg
     4441 gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat ggcagccgta
     4501 cttttatcaa gtacccgaaa ggcattcctg atttctttaa acagtccttt cctgagggtt
     4561 ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc atgcaggaca
     4621 ccagccttga ggatggctgt ctcgtttacc acgtccaagt cagaggggta aactttccct
     4681 ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca gagatgatgt
     4741 atccagcaga tggtggtctg aggggataca ctcatatggc actgaaagtt gatggtggtg
     4801 gccatctgtc ttgctctttc gtaacaactt acaggtcaaa aaagaccgtc gggaacatca
     4861 agatgcccgg tatccatgcc gttgatcacc gcctggaaag gttagaggaa agtgacaatg
     4921 aaatgttcgt agtacaacgc gaacacgcag ttgccaagtt cgccgggctt ggtggtggga
     4981 tggacgagct gtacaagtaa tctagagttg aaccgactag taagccaggt cgagacatgt
     5041 ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata atcaacctct
     5101 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     5161 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     5221 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     5281 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     5341 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     5401 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     5461 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc
     5521 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     5581 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     5641 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc gacctcgatc
     5701 gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc aatgctgatt
     5761 gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca cctcaggtac
     5821 ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta aaagaaaagg
     5881 ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat ctgtggatct
     5941 accacacaca aggctacttc cctgattggc agaactacac accagggcca gggatcagat
     6001 atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag aaggtagaag
     6061 aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat gggatggatg
     6121 acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt catcacatgg
     6181 cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6241 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6301 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6361 tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa aaggccagga
     6421 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     6481 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     6541 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     6601 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     6661 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6721 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6781 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6841 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6901 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6961 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     7021 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7081 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7141 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7201 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7261 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7321 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7381 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7441 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7501 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7561 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7621 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     7681 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     7741 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     7801 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     7861 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     7921 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     7981 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8041 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8101 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8161 taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
