LOCUS       3xFLAG-dCas9_pMX       10444 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Retroviral vector with a G418 resistance marker, encoding 
            FLAG(R)-tagged dCas9 for engineered ChIP (enChIP) purification of 
            specific genomic regions.
ACCESSION   .
VERSION     .
KEYWORDS    3xFLAG-dCas9 pMXs-neo
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10444)
  AUTHORS   Fujita T, Fujii H.
  TITLE     Identification of proteins associated with an IFN-gamma-responsive 
            promoter by a retroviral expression system for enChIP using CRISPR.
  JOURNAL   PLoS ONE 2014;9:e103084.
  PUBMED    25051498
REFERENCE   2  (bases 1 to 10444)
  AUTHORS   Fujii Lab / Addgene #51260
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10444)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2014"; volume: "9"; pages: "e103084"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10444
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..615
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     misc_feature    678..1035
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus
                     (MMLV)"
     CDS             1094..1510
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
     misc_feature    1520..1894
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     CDS             1948..1950
                     /codon_start=1
                     /product="start codon"
                     /label=start codon
                     /note="ATG"
                     /translation="M"
     CDS             1951..2016
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     CDS             2020..6123
                     /label=dCas9
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     CDS             6136..6156
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     promoter        6341..6670
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             7027..7818
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     LTR             7889..8482
                     /label=LTR
                     /note="long terminal repeat from Moloney murine leukemia
                     virus"
     rep_origin      complement(8719..9307)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9474..10331)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(10332..10436)
                     /label=AmpR promoter
ORIGIN
        1 cccgaaaagt gccacctgca taatgaaaga ccccacctgt aggtttggca agctagctta
       61 agtaacgcca ttttgcaagg catggaaaaa tacataactg agaatagaaa agttcagatc
      121 aaggtcagga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
      181 ttcctgcccc ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat
      241 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
      301 gtccagccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
      361 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
      421 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
      481 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
      541 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
      601 cgggggtctt tcatttgggg gctcgtccgg gatcgggaga cccctgccca gggaccaccg
      661 acccaccacc gggaggtaag ctggccagca acttatctgt gtctgtccga ttgtctagtg
      721 tctatgactg attttatgcg cctgcgtcgg tactagttag ctaactagct ctgtatctgg
      781 cggacccgtg gtggaactga cgagttcgga acacccggcc gcaaccctgg gagacgtccc
      841 agggacttcg ggggccgttt ttgtggcccg acctgagtcc aaaaatcccg atcgttttgg
      901 actctttggt gcacccccct aataggaggg atatgtggtt ctggtaggag acgagaacct
      961 aaaacagttc ccgcctccgt ctgaattttt gctttcggtt tgggaccgaa gccgcgccgc
     1021 gcgtcttgtc tgctgcagca tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt
     1081 gtctgaaaat tagggccaga ctgttaccac tcccttaagt ttgaccttag gtcactggaa
     1141 agatgtcgag cggatcgctc acaaccagtc ggtagatgtc aagaagagac gttgggttac
     1201 cttctgctct gcagaatggc caacctttaa cgtcggatgg ccgcgagacg gcacctttaa
     1261 ccgagacctc atcacccagg ttaagatcaa ggtcttttca cctggcccgc atggacaccc
     1321 agaccaggtc ccctacatcg tgacctggga agccttggct tttgaccccc ctccctgggt
     1381 caagcccttt gtacacccta agcctccgcc tcctcttcct ccatccgccc cgtctctccc
     1441 ccttgaacct cctcgttcga ccccgcctcg atcctccctt tatccagccc tcactccttc
     1501 tctaggcgcc cccatatggc catatgagat cttatatggg gcacccccgc cccttgtaaa
     1561 cttccctgac cctgacatga caagagttac taacagcccc tctctccaag ctcacttaca
     1621 ggctctctac ttagtccagc acgaagtctg gagacctctg gcggcagcct accaagaaca
     1681 actggaccga ccggtggtac ctcaccctta ccgagtcggc gacacagtgt gggtccgccg
     1741 acaccagact aagaacctag aacctcgctg gaaaggacct tacacagtcc tgctgaccac
     1801 ccccaccgcc ctcaaagtag acggcatcgc agcttggata cacgccgccc acgtgaaggc
     1861 tgccgacccc gggggtggac catcctctag actgccggat ctagctagtt aattaaggat
     1921 ccgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
     1981 gatcatgaca tcgattacaa ggatgacgat gacaagctca tggacaagaa gtactccatt
     2041 gggctcgcta tcggcacaaa cagcgtcggc tgggccgtca ttacggacga gtacaaggtg
     2101 ccgagcaaaa aattcaaagt tctgggcaat accgatcgcc acagcataaa gaagaacctc
     2161 attggcgccc tcctgttcga ctccggggag acggccgaag ccacgcggct caaaagaaca
     2221 gcacggcgca gatatacccg cagaaagaat cggatctgct acctgcagga gatctttagt
     2281 aatgagatgg ctaaggtgga tgactctttc ttccataggc tggaggagtc ctttttggtg
     2341 gaggaggata aaaagcacga gcgccaccca atctttggca atatcgtgga cgaggtggcg
     2401 taccatgaaa agtacccaac catatatcat ctgaggaaga agcttgtaga cagtactgat
     2461 aaggctgact tgcggttgat ctatctcgcg ctggcgcata tgatcaaatt tcggggacac
     2521 ttcctcatcg agggggacct gaacccagac aacagcgatg tcgacaaact ctttatccaa
     2581 ctggttcaga cttacaatca gcttttcgaa gagaacccga tcaacgcatc cggagttgac
     2641 gccaaagcaa tcctgagcgc taggctgtcc aaatcccggc ggctcgaaaa cctcatcgca
     2701 cagctccctg gggagaagaa gaacggcctg tttggtaatc ttatcgccct gtcactcggg
     2761 ctgaccccca actttaaatc taacttcgac ctggccgaag atgccaagct tcaactgagc
     2821 aaagacacct acgatgatga tctcgacaat ctgctggccc agatcggcga ccagtacgca
     2881 gacctttttt tggcggcaaa gaacctgtca gacgccattc tgctgagtga tattctgcga
     2941 gtgaacacgg agatcaccaa agctccgctg agcgctagta tgatcaagcg ctatgatgag
     3001 caccaccaag acttgacttt gctgaaggcc cttgtcagac agcaactgcc tgagaagtac
     3061 aaggaaattt tcttcgatca gtctaaaaat ggctacgccg gatacattga cggcggagca
     3121 agccaggagg aattttacaa atttattaag cccatcttgg aaaaaatgga cggcaccgag
     3181 gagctgctgg taaagcttaa cagagaagat ctgttgcgca aacagcgcac tttcgacaat
     3241 ggaagcatcc cccaccagat tcacctgggc gaactgcacg ctatcctcag gcggcaagag
     3301 gatttctacc cctttttgaa agataacagg gaaaagattg agaaaatcct cacatttcgg
     3361 ataccctact atgtaggccc cctcgcccgg ggaaattcca gattcgcgtg gatgactcgc
     3421 aaatcagaag agaccatcac tccctggaac ttcgaggaag tcgtggataa gggggcctct
     3481 gcccagtcct tcatcgaaag gatgactaac tttgataaaa atctgcctaa cgaaaaggtg
     3541 cttcctaaac actctctgct gtacgagtac ttcacagttt ataacgagct caccaaggtc
     3601 aaatacgtca cagaagggat gagaaagcca gcattcctgt ctggagagca gaagaaagct
     3661 atcgtggacc tcctcttcaa gacgaaccgg aaagttaccg tgaaacagct caaagaagac
     3721 tatttcaaaa agattgaatg tttcgactct gttgaaatca gcggagtgga ggatcgcttc
     3781 aacgcatccc tgggaacgta tcacgatctc ctgaaaatca ttaaagacaa ggacttcctg
     3841 gacaatgagg agaacgagga cattcttgag gacattgtcc tcacccttac gttgtttgaa
     3901 gatagggaga tgattgaaga acgcttgaaa acttacgctc atctcttcga cgacaaagtc
     3961 atgaaacagc tcaagaggcg ccgatataca ggatgggggc ggctgtcaag aaaactgatc
     4021 aatgggatcc gagacaagca gagtggaaag acaatcctgg attttcttaa gtccgatgga
     4081 tttgccaaca ggaacttcat gcagttgatc catgatgact ctctcacctt taaggaggac
     4141 atccagaaag cacaagtttc tggccagggg gacagtcttc acgagcacat cgctaatctt
     4201 gcaggtagcc cagctatcaa aaagggaata ctgcagaccg ttaaggtcgt ggatgaactc
     4261 gtcaaagtaa tgggaaggca taagcccgag aatatcgtta tcgagatggc ccgagagaac
     4321 caaactaccc agaagggaca gaagaacagt agggaaagga tgaagaggat tgaagagggt
     4381 ataaaagaac tggggtccca aatccttaag gaacacccag ttgaaaacac ccagcttcag
     4441 aatgagaagc tctacctgta ctacctgcag aacggcaggg acatgtacgt ggatcaggaa
     4501 ctggacatca atcggctctc cgactacgac gtggatgcca tcgtgcccca gtcttttctc
     4561 aaagatgatt ctattgataa taaagtgttg acaagatccg ataaaaatag agggaagagt
     4621 gataacgtcc cctcagaaga agttgtcaag aaaatgaaaa attattggcg gcagctgctg
     4681 aacgccaaac tgatcacaca acggaagttc gataatctga ctaaggctga acgaggtggc
     4741 ctgtctgagt tggataaagc cggcttcatc aaaaggcagc ttgttgagac acgccagatc
     4801 accaagcacg tggcccaaat tctcgattca cgcatgaaca ccaagtacga tgaaaatgac
     4861 aaactgattc gagaggtgaa agttattact ctgaagtcta agctggtctc agatttcaga
     4921 aaggactttc agttttataa ggtgagagag atcaacaatt accaccatgc gcatgatgcc
     4981 tacctgaatg cagtggtagg cactgcactt atcaaaaaat atcccaagct tgaatctgaa
     5041 tttgtttacg gagactataa agtgtacgat gttaggaaaa tgatcgcaaa gtctgagcag
     5101 gaaataggca aggccaccgc taagtacttc ttttacagca atattatgaa ttttttcaag
     5161 accgagatta cactggccaa tggagagatt cggaagcgac cacttatcga aacaaacgga
     5221 gaaacaggag aaatcgtgtg ggacaagggt agggatttcg cgacagtccg gaaggtcctg
     5281 tccatgccgc aggtgaacat cgttaaaaag accgaagtac agaccggagg cttctccaag
     5341 gaaagtatcc tcccgaaaag gaacagcgac aagctgatcg cacgcaaaaa agattgggac
     5401 cccaagaaat acggcggatt cgattctcct acagtcgctt acagtgtact ggttgtggcc
     5461 aaagtggaga aagggaagtc taaaaaactc aaaagcgtca aggaactgct gggcatcaca
     5521 atcatggagc gatcaagctt cgaaaaaaac cccatcgact ttctcgaggc gaaaggatat
     5581 aaagaggtca aaaaagacct catcattaag cttcccaagt actctctctt tgagcttgaa
     5641 aacggccgga aacgaatgct cgctagtgcg ggcgagctgc agaaaggtaa cgagctggca
     5701 ctgccctcta aatacgttaa tttcttgtat ctggccagcc actatgaaaa gctcaaaggg
     5761 tctcccgaag ataatgagca gaagcagctg ttcgtggaac aacacaaaca ctaccttgat
     5821 gagatcatcg agcaaataag cgaattctcc aaaagagtga tcctcgccga cgctaacctc
     5881 gataaggtgc tttctgctta caataagcac agggataagc ccatcaggga gcaggcagaa
     5941 aacattatcc acttgtttac tctgaccaac ttgggcgcgc ctgcagcctt caagtacttc
     6001 gacaccacca tagacagaaa gcggtacacc tctacaaagg aggtcctgga cgccacactg
     6061 attcatcagt caattacggg gctctatgaa acaagaatcg acctctctca gctcggtgga
     6121 gacagcaggg ctgaccccaa gaagaagagg aaggtgtgaa agggttcgat ccctaccgga
     6181 tcgaattcca gctgagcgcc ggtcgctacc attaccagtt ggtctggtgt caaaaataat
     6241 aataaccggg caggccatgt ctgcccgtat ttcgcgtaag gaaatccatt atgtactatt
     6301 taaactcgag cggccgccag cacagtggtc gatgtggaat gtgtgtcagt tagggtgtgg
     6361 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6421 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6481 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6541 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     6601 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     6661 cttttgcaaa aagcttcacg ctgccgcaag cactcagggc gcaagggctg ctaaaggaag
     6721 cggaacacgt agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac
     6781 tgggctatct ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg
     6841 cttacatggc gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca
     6901 gctggggcgc cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg
     6961 ccgccaagga tctgatggcg caggggatca agatctgatc aagagacagg atgaggatcg
     7021 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     7081 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     7141 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     7201 gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     7261 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     7321 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     7381 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     7441 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     7501 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg
     7561 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     7621 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     7681 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     7741 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     7801 cttcttgacg agttcttctg agcgggactc tggggttatc gtcgacgata aaataaaaga
     7861 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     7921 tagcttaagt aacgccattt tggaaggcat ggaaaaatac ataactgaga atagagaagt
     7981 tcagatcaag gtcaggaaca gatggaacag ctgaatatgg gccaaacagg atatctgtgg
     8041 taagcagttc ctgccccggc tcagggccaa gaacagatgg aacagctgaa tatgggccaa
     8101 acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca gatggtcccc
     8161 agatgcggtc cagccctcag cagtttctag agaaccatca gatgtttcca gggtgcccca
     8221 aggacctgaa atgaccctgt gccttatttg aactaaccaa tcagttcgct tctcgcttct
     8281 gttcgcgcgc ttctgctccc cgagctcaat aaaagagccc acaacccctc actcggggcg
     8341 ccagtcctcc gattgactga gtcgcccggg tacccgtgta tccaataaac cctcttgcag
     8401 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
     8461 ccgtcagcgg gggtctttca ttctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     8521 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     8581 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     8641 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     8701 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     8761 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     8821 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     8881 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     8941 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     9001 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     9061 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     9121 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
     9181 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     9241 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     9301 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     9361 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttgcg
     9421 gccggccgca aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     9481 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     9541 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     9601 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     9661 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     9721 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     9781 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     9841 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     9901 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     9961 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
    10021 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
    10081 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
    10141 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
    10201 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
    10261 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
    10321 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
    10381 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
    10441 tttc
//
