LOCUS       sequence-356956-        6352 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Plasmid for expression of FLAG-GFP tagged human FUS/TLS (C-terminal 
            tag). Confers resistance to G418..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6352)
  AUTHORS   Britton S, Dernoncourt E, Delteil C, Froment C, Schiltz O, Salles B,
            Frit P, Calsou P
  TITLE     DNA damage triggers SAF-A and RNA biogenesis factors exclusion from 
            chromatin coupled to R-loops removal.
  JOURNAL   Nucleic Acids Res. 2014 Oct 1;42(14):9047-62. doi: 
            10.1093/nar/gku601. Epub 2014 Jul 16.
  PUBMED    25030905
REFERENCE   2  (bases 1 to 6352)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1093/nar/gku601"; journalName: "Nucleic Acids Res"; date: 
            "2014-10-1- 1"; volume: "42"; issue: "14"; pages: "9047-62"
COMMENT     Sequence Label: pEGFP-N1-FUS/TLS-FLAGC
FEATURES             Location/Qualifiers
     source          1..6352
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             6..29
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      39..48
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(45..47,48..50,51..764)
                     /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             join(45..47,48..50,51..761)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(90..111)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(351..370)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     698..719
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    887..1008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(924..943)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     978..997
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1015..1470)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1152..1173)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1364..1383
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1497..1601
                     /gene="bla"
                     /label=AmpR promoter
     promoter        1603..1960
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        1603..1794
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(1626..1646)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1811..1946
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1995..2789
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             1995..2789
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2049..2068)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2659..2678
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2977..2996)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3021..3068
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3397..3985
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3886..3905
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4160..4463
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4464..4667
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4618..4638
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    4708..4764
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      4768..4777
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5154..5165
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
ORIGIN
        1 cgcgtgatta caaggatgac gacgataagc caccggtcgc caccatggtg agcaagggcg
       61 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      121 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      181 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      241 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      301 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
      361 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      421 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
      481 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
      541 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      601 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
      661 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      721 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
      781 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      841 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      901 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      961 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     1021 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     1081 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     1141 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     1201 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     1261 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     1321 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     1381 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     1441 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     1501 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1561 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     1621 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     1681 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     1741 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     1801 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     1861 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     1921 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     1981 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     2041 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     2101 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     2161 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     2221 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     2281 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     2341 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     2401 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     2461 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     2521 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     2581 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     2641 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     2701 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     2761 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     2821 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     2881 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     2941 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     3001 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     3061 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     3121 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     3181 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     3241 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     3301 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     3361 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     3421 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     3481 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     3541 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     3601 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     3661 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3721 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3781 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3841 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3901 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3961 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     4021 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     4081 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
     4141 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     4201 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     4261 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     4321 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     4381 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     4441 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     4501 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     4561 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     4621 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     4681 gtcagatccg ctagcgctac cggactcaga tctcgagctc aagcttcgaa ttctgcagtc
     4741 gacggtaccg cgggcccggg atccgccgcc accatggcct caaacgatta tacccaacaa
     4801 gcaacccaaa gctatggggc ctaccccacc cagcccgggc agggctattc ccagcagagc
     4861 agtcagccct acggacagca gagttacagt ggttatagcc agtccacgga cacttcaggc
     4921 tatggccaga gcagctattc ttcttatggc cagagccaga acacaggcta tggaactcag
     4981 tcaactcccc agggatatgg ctcgactggc ggctatggca gtagccagag ctcccaatcg
     5041 tcttacgggc agcagtcctc ctatcctggc tatggccagc agccagctcc cagcagcacc
     5101 tcgggaagtt acggtagcag ttctcagagc agcagctatg ggcagcccca gagtgggagc
     5161 tacagccagc agcctagcta tggtggacag cagcaaagct atggacagca gcaaagctat
     5221 aatccccctc agggctatgg acagcagaac cagtacaaca gcagcagtgg tggtggaggt
     5281 ggaggtggag gtggaggtaa ctatggccaa gatcaatcct ccatgagtag tggtggtggc
     5341 agtggtggcg gttatggcaa tcaagaccag agtggtggag gtggcagcgg tggctatgga
     5401 cagcaggacc gtggaggccg cggcaggggt ggcagtggtg gcggcggcgg cggcggcggt
     5461 ggtggttaca accgcagcag tggtggctat gaacccagag gtcgtggagg tggccgtgga
     5521 ggcagaggtg gcatgggcgg aagtgaccgt ggtggcttca ataaatttgg tggccctcgg
     5581 gaccaaggat cacgtcatga ctccgaacag gataattcag acaacaacac catctttgtg
     5641 caaggcctgg gtgagaatgt tacaattgag tctgtggctg attacttcaa gcagattggt
     5701 attattaaga caaacaagaa aacgggacag cccatgatta atttgtacac agacagggaa
     5761 actggcaagc tgaagggaga ggcaacggtc tcttttgatg acccaccttc agctaaagca
     5821 gctattgact ggtttgatgg taaagaattc tccggaaatc ctatcaaggt ctcatttgct
     5881 actcgccggg cagactttaa tcggggtggt ggcaatggtc gtggaggccg agggcgagga
     5941 ggacccatgg gccgtggagg ctatggaggt ggtggcagtg gtggtggtgg ccgaggagga
     6001 tttcccagtg gaggtggtgg cggtggagga cagcagcgag ctggtgactg gaagtgtcct
     6061 aatcccacct gtgagaatat gaacttctct tggaggaatg aatgcaacca gtgtaaggcc
     6121 cctaaaccag atggcccagg agggggacca ggtggctctc acatgggggg taactacggg
     6181 gatgatcgtc gtggtggcag aggaggctat gatcgaggcg gctaccgggg ccgcggcggg
     6241 gaccgtggag gcttccgagg gggccggggt ggtggggaca gaggtggctt tggccctggc
     6301 aagatggatt ccaggggtga gcacagacag gatcgcaggg agaggccgta ta
//
