LOCUS       sequence-448137-        7913 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expressing Shuttle-tdTomato, or NLS-NES-tagged tdTomato in a 
            Lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7913)
  AUTHORS   Zhang K, Donnelly CJ, Haeusler AR, Grima JC, Machamer JB, Steinwald 
            P, Daley EL, Miller SJ, Cunningham KM, Vidensky S, Gupta S, Thomas 
            MA, Hong I, Chiu SL, Huganir RL, Ostrow LW, Matunis MJ, Wang J, 
            Sattler R, Lloyd TE, Rothstein JD
  TITLE     The C9orf72 repeat expansion disrupts nucleocytoplasmic transport.
  JOURNAL   Nature. 2015 Sep 3;525(7567):56-61. doi: 10.1038/nature14973. Epub 
            2015 Aug 26.
  PUBMED    26308891
REFERENCE   2  (bases 1 to 7913)
  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: 
            "10.1038/nature14973"; journalName: "Nature"; date: "2015-09-3- 3"; 
            volume: "525"; issue: "7567"; pages: "56-61"
COMMENT     Sequence Label: Lentiviral-S-tdTomato
FEATURES             Location/Qualifiers
     source          1..7913
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(6..665)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     580..599
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(666..770)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(845..864)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1035..1414
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1416..1614
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             1632..1812
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1859..1984
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    2481..2714
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2899..2943
                     /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    3241..3358
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     enhancer        3441..3820
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3821..4024
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4050..4069
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4050..4068
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4183..4203
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4219..5646
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     CDS             5653..5679
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     CDS             5683..5712
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             5731..5754
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(5806..5839)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    5895..6483
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5948..5968)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6366..6377)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(6486..6502)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6487..6503)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     LTR             7012..7192
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(7254..7842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7334..7353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgattacg ccccgccctg ccactcatcg cagtactgtt gtaattcatt aagcattctg
       61 ccgacatgga agccatcaca aacggcatga tgaacctgaa tcgccagcgg catcagcacc
      121 ttgtcgcctt gcgtataata tttgcccatg gtgaaaacgg gggcgaagaa gttgtccata
      181 ttggccacgt ttaaatcaaa actggtgaaa ctcacccagg gattggctga gacgaaaaac
      241 atattctcaa taaacccttt agggaaatag gccaggtttt caccgtaaca cgccacatct
      301 tgcgaatata tgtgtagaaa ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa
      361 aacgtttcag tttgctcatg gaaaacggtg taacaagggt gaacactatc ccatatcacc
      421 agctcaccgt ctttcattgc catacggaac tccggatgag cattcatcag gcgggcaaga
      481 atgtgaataa aggccggata aaacttgtgc ttatttttct ttacggtctt taaaaaggcc
      541 gtaatatcca gctgaacggt ctggttatag gtacattgag caactgactg aaatgcctca
      601 aaatgttctt tacgatgcca ttgggatata tcaacggtgg tatatccagt gatttttttc
      661 tccatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
      721 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
      781 cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc
      841 tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt
      901 tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg
      961 acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg
     1021 ccagatatcg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     1081 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     1141 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     1201 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     1261 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
     1321 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
     1381 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
     1441 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     1501 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
     1561 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagcgcgtt
     1621 ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac
     1681 tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg
     1741 cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga
     1801 aaatctctag cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct
     1861 ctctcgacgc aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact
     1921 ggtgagtacg ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag
     1981 cgtcagtatt aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg
     2041 gggaaagaaa aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt
     2101 cgcagttaat cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct
     2161 acaaccatcc cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac
     2221 cctctattgt gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat
     2281 agaggaagag caaaacaaaa gtaagaccac cgcacagcaa gcggccggcc gctgatcttc
     2341 agacctggag gaggagatat gagggacaat tggagaagtg aattatataa atataaagta
     2401 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga
     2461 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc
     2521 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctggtata
     2581 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc
     2641 acagtctggg gcatcaagca gctccaggca agaatcctgg ctgtggaaag atacctaaag
     2701 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg
     2761 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaatca cacgacctgg
     2821 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa
     2881 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt
     2941 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata
     3001 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt
     3061 aggcagggat attcaccatt atcgtttcag acccacctcc caaccccgag gggacccgac
     3121 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta
     3181 gtgaacggat cggcactgcg tgcgccaatt ctgcagacaa atggcagtat tcatccacaa
     3241 ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag tagacataat
     3301 agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaaattc aaaattttcg
     3361 ggtttattac agggacagca gagatccagt ttggttagta ccgggcccgc tctagacatg
     3421 tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
     3481 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     3541 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     3601 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     3661 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     3721 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     3781 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     3841 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     3901 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     3961 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     4021 agctcgttta gtgaaccgtc agaattttgt aatacgactc actatagggc ggccgggaat
     4081 tcgtcgactg gatccggtac cgaggagatc tgccgccgcg atcgccggcg cgccagatct
     4141 caagcttaac tagctagcgg accgacgcgt gccaccatgc ccccgaagaa aaagcggaag
     4201 gtccaaggag gcgggggcat ggtgagcaag ggcgaggagg tcatcaaaga gttcatgcgc
     4261 ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga gggcgagggc
     4321 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
     4381 ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa ggcgtacgtg
     4441 aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg cttcaagtgg
     4501 gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga ctcctccctg
     4561 caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc ccccgacggc
     4621 cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct gtacccccgc
     4681 gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg cggccactac
     4741 ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc cggctactac
     4801 tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     4861 tacgagcgct ccgagggccg ccaccacctg ttcctggggc atggcaccgg cagcaccggc
     4921 agcggcagct ccggcaccgc ctcctccgag gacaacaaca tggccgtcat caaagagttc
     4981 atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga gatcgagggc
     5041 gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt gaccaagggc
     5101 ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg ctccaaggcg
     5161 tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc cgagggcttc
     5221 aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac ccaggactcc
     5281 tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa cttccccccc
     5341 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga gcgcctgtac
     5401 ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa ggacggcggc
     5461 cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca actgcccggc
     5521 tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta caccatcgtg
     5581 gaacagtacg agcgctccga gggccgccac cacctgttcc tgtacggcat ggacgagctg
     5641 tacaagctac agctaccgcc gctggaaaga ctgactctgc tcgagcagaa actcatctca
     5701 gaagaggatc tggcagcaaa tgatatcctg gattacaagg atgacgacga taaggtttaa
     5761 acggccggcc gcggtctgta caagtaggat tcgtcgaggg acctaataac ttcgtatagc
     5821 atacattata cgaagttata catgtttaag ggttccggtt ccactaggta caattcgata
     5881 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     5941 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     6001 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     6061 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     6121 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     6181 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     6241 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     6301 ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     6361 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     6421 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     6481 cgcatcgata ccgtcgacct cgatcgagac ctagaaaaac atggagcaat cacaagtagc
     6541 aatacagcag ctaccaatgc tgattgtgcc tggctagaag cacaagagga ggaggaggtg
     6601 ggttttccag tcacacctca ggtaccttta agaccaatga cttacaaggc agctgtagat
     6661 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaga
     6721 caagatatcc ttgatctgtg gatctaccac acacaaggct acttccctga ttggcagaac
     6781 tacacaccag ggccagggat cagatatcca ctgacctttg gatggtgcta caagctagta
     6841 ccagttgagc aagagaaggt agaagaagcc aatgaaggag agaacacccg cttgttacac
     6901 cctgtgagcc tgcatgggat ggatgacccg gagagagaag tattagagtg gaggtttgac
     6961 agccgcctag catttcatca catggcccga gagctgcatc cggactgtac tgggtctctc
     7021 tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc actgcttaag
     7081 cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct
     7141 ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag cagcatgtga
     7201 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7441 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7681 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7801 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggt
//
