LOCUS       EF1a-FLuc-F2A-GF        7713 bp    DNA     linear        8-JUL-2013
SOURCE      
  ORGANISM  
            This file is created by Biosettia Inc
            http://www.biosettia.com/
FEATURES             Location/Qualifiers
     LTR             7479..7713
                     /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             6191..6790
                     /label=Puro
     RBS             5583..6167
                     /label=IRES
     misc_feature    6800..7396
                     /label=WPRE
     CDS             4793..4858
                     /label=F2A
     CDS             4859..5578
                     /label=GFP
     CDS             3128..4777
                     /label=Firefly\Luciferase\(FLuc)
BASE COUNT     1798 a      2125 c      2188 g      1602 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 tggatccatg gaagatgcca aaaacattaa gaagggccca gcgccattct acccactcga 
     3181 agacgggacc gccggcgagc agctgcacaa agccatgaag cgctacgccc tggtgcccgg 
     3241 caccatcgcc tttaccgacg cacatatcga ggtggacatt acctacgccg agtacttcga 
     3301 gatgagcgtt cggctggcag aagctatgaa gcgctatggg ctgaatacaa accatcggat 
     3361 cgtggtgtgc agcgagaata gcttgcagtt cttcatgccc gtgttgggtg ccctgttcat 
     3421 cggtgtggct gtggccccag ctaacgacat ctacaacgag cgcgagctgc tgaacagcat 
     3481 gggcatcagc cagcccaccg tcgtattcgt gagcaagaaa gggctgcaaa agatcctcaa 
     3541 cgtgcaaaag aagctaccga tcatacaaaa gatcatcatc atggatagca agaccgacta 
     3601 ccagggcttc caaagcatgt acaccttcgt gacttcccat ttgccacccg gcttcaacga 
     3661 gtacgacttc gtgcccgaga gcttcgaccg ggacaaaacc atcgccctga tcatgaacag 
     3721 tagtggcagt accggattgc ccaagggcgt agccctaccg caccgcaccg cttgtgtccg 
     3781 attcagtcat gcccgcgacc ccatcttcgg caaccagatc atccccgaca ccgctatcct 
     3841 cagcgtggtg ccatttcacc acggcttcgg catgttcacc acgctgggct acttgatctg 
     3901 cggctttcgg gtcgtgctca tgtaccgctt cgaggaggag ctattcttgc gcagcttgca 
     3961 agactataag attcaatctg ccctgctggt gcccacacta tttagcttct tcgctaagag 
     4021 cactctcatc gacaagtacg acctaagcaa cttgcacgag atcgccagcg gcggggcgcc 
     4081 gctcagcaag gaggtaggtg aggccgtggc caaacgcttc cacctaccag gcatccgcca 
     4141 gggctacggc ctgacagaaa caaccagcgc cattctgatc acccccgaag gggacgacaa 
     4201 gcctggcgca gtaggcaagg tggtgccctt cttcgaggct aaggtggtgg acttggacac 
     4261 cggtaagaca ctgggtgtga accagcgcgg cgagctgtgc gtccgtggcc ccatgatcat 
     4321 gagcggctac gttaacaacc ccgaggctac aaacgctctc atcgacaagg acggctggct 
     4381 gcacagcggc gacatcgcct actgggacga ggacgagcac ttcttcatcg tggaccggct 
     4441 gaagagcctg atcaaataca agggctacca ggtagcccca gccgaactgg agagcatcct 
     4501 gctgcaacac cccaacatct tcgacgccgg ggtcgccggc ctgcccgacg acgatgccgg 
     4561 cgagctgccc gccgcagtcg tcgtgctgga acacggtaaa accatgaccg agaaggagat 
     4621 cgtggactat gtggccagcc aggttacaac cgccaagaag ctgcgcggtg gtgttgtgtt 
     4681 cgtggacgag gtgcctaaag gactgaccgg caagttggac gcccgcaaga tccgcgagat 
     4741 tctcattaag gccaagaagg gcggcaagat cgccgtggct agcggcgata tcgtgaagca 
     4801 gaccctgaac ttcgacctcc tgaagctggc cggcgacgtg gaaagcaacc ccgggcctat 
     4861 ggtgtccaag ggcgaggaac tgttcaccgg cgtggtgccc atcctggtgg aactggatgg 
     4921 cgacgtgaac ggccacaagt tcagcgtgtc cggcgagggc gaaggcgacg ccacatatgg 
     4981 caagctgaca ctgaagttca tctgcaccac cggcaagctg cccgtgcctt ggcctaccct 
     5041 cgtgaccaca ctgacctacg gcgtgcagtg cttctccaga taccccgacc atatgaagca 
     5101 gcacgacttc ttcaagagcg ccatgcccga gggctacgtg caggaacgga ccatcttctt 
     5161 taaggacgac ggcaactaca agaccagggc cgaagtgaag ttcgagggcg acaccctcgt 
     5221 gaaccggatc gagctgaagg gcatcgactt caaagaggac ggcaacatcc tgggccacaa 
     5281 gctggagtac aactacaaca gccacaacgt ctacatcatg gccgacaagc agaaaaacgg 
     5341 catcaaagtg aacttcaaga tccggcacaa catcgaggac ggctccgtgc agctggccga 
     5401 ccactaccag cagaacaccc ccatcggaga tggccccgtg ctgctgcccg acaaccacta 
     5461 cctgagcaca cagagcgccc tgagcaagga ccccaacgag aagcgggacc acatggtgct 
     5521 gctggagttc gtgaccgccg ctggcatcac cctgggcatg gacgagctgt acaagtgaac 
     5581 tagcccctct ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg 
     5641 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc 
     5701 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa 
     5761 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag 
     5821 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg 
     5881 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg 
     5941 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa 
     6001 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg 
     6061 gtacacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca 
     6121 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacacta gagatccacc 
     6181 ggtcgccacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc 
     6241 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt 
     6301 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt 
     6361 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac 
     6421 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga 
     6481 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg 
     6541 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa 
     6601 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc 
     6661 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac 
     6721 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc 
     6781 cggtgcctga gcggccgcgt cgacaatcaa cctctggatt acaaaatttg tgaaagattg 
     6841 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 
     6901 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 
     6961 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 
     7021 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 
     7081 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 
     7141 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag 
     7201 ctgacgtcct ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 
     7261 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 
     7321 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 
     7381 gccgcctccc cgcctggaat tcgagctcgg tacctttaag accaatgact tacaaggcag 
     7441 ctgtagatct tagccacttt ttaaaagaaa aggggggact ggaagggcta attcactccc 
     7501 aacgaagaca agatctgctt tttgcttgta ctgggtctct ctggttagac cagatctgag 
     7561 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt 
     7621 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca 
     7681 gaccctttta gtcagtgtgg aaaatctcta gca 
//
