LOCUS       Exported                9411 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Lentiviral vector with a (Ubiquitous Chromatin Opening Element) UCOE
            upstream of a CAG promoter constitutively expressing a single insert
            containing the Tau.K18(P301L/V337M)-mScarlet-I fusion protein..
ACCESSION   .
VERSION     .
KEYWORDS    pJC49
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9411)
  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 9411)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9411
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     enhancer        3402..3705
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3707..3982
                     /label=chicken beta-actin promoter
     intron          3983..4999
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     4923..4945
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     5007..5026
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             5498..6190
                     /codon_start=1
                     /product="bright monomeric red fluorescent protein evolved 
                     from a synthetic template (Bindels et al., 2016)"
                     /label=mScarlet-I
                     /note="matures faster than mScarlet"
                     /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFSWDILSPQFMYGSRAFIKHPADIPDYYKQSFPEGFKWERVMNFEDGGA
                     VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM
                     ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS
                     TGGMDELYK"
     primer_bind     complement(5628..5646)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     5896..5915
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    6300..6888
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(6353..6373)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6771..6782)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6891..6907)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6892..6908)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7417..7597
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7659..8247)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7739..7758)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(8418..9278)
                     /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     9041..9060
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9279..9383)
                     /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 tcgggaggtg gtccctgcag ttacgccaat gataaccccc gccagaaaaa
     2701 tcttagtagc cttccctttt tgttttccgt gccccaactc ggcggattga ctcggcccct
     2761 tccggaaaca cccgaatcaa cttctagtca aattattgtt cacgccgcaa tgacccaccc
     2821 ctggcccgcg tctgtggaac tgacccctgg tgtacaggag agttcgctgc tgaaagtggt
     2881 cccaaagggg tactagtttt taagctccca actccccctc ccccagcgtc tggaggattc
     2941 cacaccctcg caccgcaggg gcgaggaagt gggcggagtc cggttttggc gccagccgct
     3001 gaggctgcca agcagaaaag ccaccgctga ggagactccg gtcactgtcc tcgccccgcc
     3061 tcccccttcc ctccccttgg ggaccaccgg gcgccacgcc gcgaacggta agtgccgcgg
     3121 tcgtcggcgc ctccgccctc cccctagggc cccaattccc agcgggcgcg gcgcgcggcc
     3181 cctccccccg ccgggcgcgc gcccgctgcc ccgcccttcg tggccgcccg gcgtgggcgg
     3241 tgccacccct ccccccggcg gccccgcgcg cagctcccgg ctccctcccc cttcggatgt
     3301 ggcttgagct gtaggcgcgg agatgcatat tgattattga ctagttatta atagtaatca
     3361 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     3421 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     3481 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     3541 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     3601 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     3661 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtcga ggtgagcccc
     3721 acgttctgct tcactctccc catctccccc ccctccccac ccccaatttt gtatttattt
     3781 attttttaat tattttgtgc agcgatgggg gcgggggggg ggggggsgcg cgccaggcgg
     3841 ggcggggcgg ggcgaggggc ggggtggggc gaggcggaga ggtgcggcgg cagccaatca
     3901 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa
     3961 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc
     4021 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
     4081 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
     4141 ttcttttctg tgactgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
     4201 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc
     4261 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt
     4321 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa
     4381 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt
     4441 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg
     4501 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca
     4561 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg
     4621 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt
     4681 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa
     4741 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg
     4801 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc
     4861 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
     4921 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     4981 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     5041 tggcaaagaa ttcctcgagc tagccaccat gctgcagaca gcccccgtgc ccatgccaga
     5101 cctgaagaat gtcaagtcca agatcggctc cactgagaac ctgaagcacc agccgggagg
     5161 cgggaaggtg cagataatta ataagaagct ggatcttagc aacgtccagt ccaagtgtgg
     5221 ctcaaaggat aatatcaaac acgtcctggg aggcggcagt gtgcaaatag tctacaaacc
     5281 agttgacctg agcaaggtga cctccaagtg tggctcatta ggcaacatcc atcataaacc
     5341 aggaggtggc cagatggaag taaaatctga gaagcttgac ttcaaggaca gagtccagtc
     5401 gaagattggg tccctggaca atatcaccca cgtccctggc ggaggaaata aaaagattga
     5461 aacccacaag tctagtggta gtagcggccg cagtagcgtg agcaagggcg aggcagtgat
     5521 caaggagttc atgcggttca aggtgcacat ggagggctcc atgaacggcc acgagttcga
     5581 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     5641 gaccaagggt ggccccctgc ccttctcctg ggacatcctg tcccctcagt tcatgtacgg
     5701 ctccagggcc ttcatcaagc accccgccga catccccgac tactataagc agtccttccc
     5761 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgccg tgaccgtgac
     5821 ccaggacacc tccctggagg acggcaccct gatctacaag gtgaagctcc gcggcaccaa
     5881 cttccctcct gacggccccg taatgcagaa gaagacaatg ggctgggaag cgtccaccga
     5941 gcggttgtac cccgaggacg gcgtgctgaa gggcgacatt aagatggccc tgcgcctgaa
     6001 ggacggcgga cgctacctgg cggacttcaa gaccacctac aaggccaaga agcccgtgca
     6061 gatgcccggc gcctacaacg tcgaccgcaa gttggacatc acctcccaca acgaggacta
     6121 caccgtggtg gaacagtacg aacgctccga gggccgccac tccaccggcg gcatggacga
     6181 gctgtacaag taagtttaaa cgcgatcgca cctagagttg aaccgactag taagccaggt
     6241 cgagacatgt ttaagggttc cggttccact aggtacaatt cgatatcaag cttatcgata
     6301 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     6361 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     6421 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     6481 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     6541 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     6601 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     6661 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     6721 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6781 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6841 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgtc
     6901 gacctcgatc gagacctaga aaaacatgga gcaatcacaa gtagcaatac agcagctacc
     6961 aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt tccagtcaca
     7021 cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag ccacttttta
     7081 aaagaaaagg ggggactgga agggctaatt cactcccaac gaagacaaga tatccttgat
     7141 ctgtggatct accacacaca aggctacttc cctgattggc agaactacac accagggcca
     7201 gggatcagat atccactgac ctttggatgg tgctacaagc tagtaccagt tgagcaagag
     7261 aaggtagaag aagccaatga aggagagaac acccgcttgt tacaccctgt gagcctgcat
     7321 gggatggatg acccggagag agaagtatta gagtggaggt ttgacagccg cctagcattt
     7381 catcacatgg cccgagagct gcatccggac tgtactgggt ctctctggtt agaccagatc
     7441 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7501 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7561 ctcagaccct tttagtcagt gtggaaaatc tctagcagca tgtgagcaaa aggccagcaa
     7621 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7681 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7741 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     7801 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     7861 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     7921 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     7981 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8041 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     8101 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8161 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8221 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8281 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8341 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8401 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8461 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8521 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8581 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8641 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8701 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     8761 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     8821 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     8881 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     8941 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9001 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9061 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9121 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9181 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9241 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9301 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9361 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga c
//
