LOCUS       EF1a-RLuc-F2A-GF        6999 bp    DNA     linear        8-JUL-2013
SOURCE      
  ORGANISM  
            This file is created by Biosettia Inc
            http://www.biosettia.com/
FEATURES             Location/Qualifiers
     LTR             6765..6999
                     /label=3'\LTR
     intron          291..1455
                     /label=Intron
     LTR             1..181
                     /label=5'LTR
     promoter        1698..3087
                     /label=human\EF1a\promoter
     intron          2126..3068
                     /label=intron
     CDS             5477..6076
                     /label=Puro
     RBS             4869..5453
                     /label=IRES
     misc_feature    6086..6682
                     /label=WPRE
     CDS             4079..4144
                     /label=F2A
     CDS             4145..4864
                     /label=GFP
     CDS             3134..4063
                     /label=Renilla\Luciferase\(RLuc)
BASE COUNT     1630 a      1885 c      1988 g      1496 t 
ORIGIN
        1 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca 
       61 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg 
      121 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc 
      181 agtggcgccc gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca 
      241 ggactcggct tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc 
      301 caaaaatttt gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta 
      361 agcgggggag aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa 
      421 aatataaatt aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc 
      481 ctggcctgtt agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc 
      541 ttcagacagg atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg 
      601 tgcatcaaag gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc 
      661 aaaacaaaag taagaccacc gcacagcaag cggccgctga tcttcagacc tggaggagga 
      721 gatatgaggg acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca 
      781 ttaggagtag cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg 
      841 ggaataggag ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcg 
      901 tcaatgacgc tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac 
      961 aatttgctga gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc 
     1021 aagcagctcc aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg 
     1081 gggatttggg gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt 
     1141 tggagtaata aatctctgga acagatttgg aatcacacga cctggatgga gtgggacaga 
     1201 gaaattaaca attacacaag cttaatacac tccttaattg aagaatcgca aaaccagcaa 
     1261 gaaaagaatg aacaagaatt attggaatta gataaatggg caagtttgtg gaattggttt 
     1321 aacataacaa attggctgtg gtatataaaa ttattcataa tgatagtagg aggcttggta 
     1381 ggtttaagaa tagtttttgc tgtactttct atagtgaata gagttaggca gggatattca 
     1441 ccattatcgt ttcagaccca cctcccaacc ccgaggggac ccgacaggcc cgaaggaata 
     1501 gaagaagaag gtggagagag agacagagac agatccattc gattagtgaa cggatctcga 
     1561 cggtatcggt taacttttaa aagaaaaggg gggattgggg ggtacagtgc aggggaaaga 
     1621 atagtagaca taatagcaac agacatacaa actaaagaat tacaaaaaca aattacaaaa 
     1681 ttcaaaattt tatcgatgcc tccccgtcac cacccccccc aacccgcccc gaccggagct 
     1741 gagagtaatt catacaaaag gactcgcccc tgccttgggg aatcccaggg accgtcgtta 
     1801 aactcccact aacgtagaac ccagagatcg ctgcgttccc gccccctcac ccgcccgctc 
     1861 tcgtcatcac tgaggtggag aagagcatgc gtgaggctcc ggtgcccgtc agtgggcaga 
     1921 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc 
     1981 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt 
     2041 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg 
     2101 caacgggttt gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct 
     2161 ctttacgggt tatggccctt gcgtgccttg aattacttcc acgcccctgg ctgcagtacg 
     2221 tgattcttga tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt 
     2281 aaggagcccc ttcgcctcgt gcttgagttg aggcctggcc tgggcgctgg ggccgccgcg 
     2341 tgcgaatctg gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt 
     2401 aaaatttttg atgatatcct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg 
     2461 gccaagatct gcacactggt atttcggttt ttggggccgc gggcggcgac ggggcccgtg 
     2521 cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc gcggccaccg agaatcggac 
     2581 gggggtagtc tcaagctggc cggcctgctc tggtgcctgg cctcgcgccg ccgtgtatcg 
     2641 ccccgccctg ggcggcaagg ctggcccggt cggcaccagt tgcgtgagcg gaaagatggc 
     2701 cgcttcccgg ccctgctgca gggagctcaa aatggaggac gcggcgctcg ggagagcggg 
     2761 cgggtgagtc acccacacaa aggaaaaggg cctttccgtc ctcagccgtc gcttcatgtg 
     2821 actccacgga gtaccgggcg ccgtccaggc acctcgatta gttctcgagc ttttggagta 
     2881 cgtcgtcttt aggttggggg gaggggtttt atgcgatgga gtttccccac actgagtggg 
     2941 tggagactga agttaggcca gcttggcact tgatgtaatt ctccttggaa tttgcccttt 
     3001 ttgagtttgg atcttggttc attctcaagc ctcagacagt ggttcaaagt ttttttcttc 
     3061 catttcaggt gtcgtgaaaa ctacccctct agataatacg actcactata gggcccgggt 
     3121 tggatccgcc accatggctt ccaaggtgta cgaccccgag caacgcaaac gcatgatcac 
     3181 tgggcctcag tggtgggctc gctgcaagca aatgaacgtg ctggactcct tcatcaacta 
     3241 ctatgattcc gagaagcacg ccgagaacgc cgtgattttt ctgcatggta acgctgcctc 
     3301 cagctacctg tggaggcacg tcgtgcctca catcgagccc gtggctagat gcatcatccc 
     3361 tgatctgatc ggaatgggta agtccggcaa gagcgggaat ggctcatatc gcctcctgga 
     3421 tcactacaag tacctcaccg cttggttcga gctgctgaac cttccaaaga aaatcatctt 
     3481 tgtgggccac gactgggggg cttgtctggc ctttcactac tcctacgagc accaagacaa 
     3541 gatcaaggcc atcgtccatg ctgagagtgt cgtggacgtg atcgagtcct gggacgagtg 
     3601 gcctgacatc gaggaggata tcgccctgat caagagcgaa gagggcgaga aaatggtgct 
     3661 tgagaataac ttcttcgtcg agaccatgct cccaagcaag atcatgcgga aactggagcc 
     3721 tgaggagttc gctgcctacc tggagccatt caaggagaag ggcgaggtta gacggcctac 
     3781 cctctcctgg cctcgcgaga tccctctcgt taagggaggc aagcccgacg tcgtccagat 
     3841 tgtccgcaac tacaacgcct accttcgggc cagcgacgat ctgcctaaga tgttcatcga 
     3901 gtccgaccct gggttctttt ccaacgctat tgtcgaggga gctaagaagt tccctaacac 
     3961 cgagttcgtg aaggtgaagg gcctccactt cagccaggag gacgctccag atgaaatggg 
     4021 taagtacatc aagagcttcg tggagcgcgt gctgaagaac gaggctagcg gcgatatcgt 
     4081 gaagcagacc ctgaacttcg acctcctgaa gctggccggc gacgtggaaa gcaaccccgg 
     4141 gcctatggtg tccaagggcg aggaactgtt caccggcgtg gtgcccatcc tggtggaact 
     4201 ggatggcgac gtgaacggcc acaagttcag cgtgtccggc gagggcgaag gcgacgccac 
     4261 atatggcaag ctgacactga agttcatctg caccaccggc aagctgcccg tgccttggcc 
     4321 taccctcgtg accacactga cctacggcgt gcagtgcttc tccagatacc ccgaccatat 
     4381 gaagcagcac gacttcttca agagcgccat gcccgagggc tacgtgcagg aacggaccat 
     4441 cttctttaag gacgacggca actacaagac cagggccgaa gtgaagttcg agggcgacac 
     4501 cctcgtgaac cggatcgagc tgaagggcat cgacttcaaa gaggacggca acatcctggg 
     4561 ccacaagctg gagtacaact acaacagcca caacgtctac atcatggccg acaagcagaa 
     4621 aaacggcatc aaagtgaact tcaagatccg gcacaacatc gaggacggct ccgtgcagct 
     4681 ggccgaccac taccagcaga acacccccat cggagatggc cccgtgctgc tgcccgacaa 
     4741 ccactacctg agcacacaga gcgccctgag caaggacccc aacgagaagc gggaccacat 
     4801 ggtgctgctg gagttcgtga ccgccgctgg catcaccctg ggcatggacg agctgtacaa 
     4861 gtgaactagc ccctctccct cccccccccc taacgttact ggccgaagcc gcttggaata 
     4921 aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt 
     4981 gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct 
     5041 cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc 
     5101 ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga 
     5161 caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc 
     5221 ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt 
     5281 attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg 
     5341 gcctcggtac acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc 
     5401 gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc acactagaga 
     5461 tccaccggtc gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga 
     5521 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca 
     5581 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac 
     5641 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt 
     5701 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat 
     5761 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc 
     5821 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca 
     5881 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg 
     5941 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg 
     6001 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg 
     6061 caagcccggt gcctgagcgg ccgcgtcgac aatcaacctc tggattacaa aatttgtgaa 
     6121 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta 
     6181 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa 
     6241 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg 
     6301 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc 
     6361 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc 
     6421 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg 
     6481 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg 
     6541 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg 
     6601 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc 
     6661 ctttgggccg cctccccgcc tggaattcga gctcggtacc tttaagacca atgacttaca 
     6721 aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa gggctaattc 
     6781 actcccaacg aagacaagat ctgctttttg cttgtactgg gtctctctgg ttagaccaga 
     6841 tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct 
     6901 tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat 
     6961 ccctcagacc cttttagtca gtgtggaaaa tctctagca 
//
