LOCUS       EF1a-FLuc-T2A-RF        8118 bp    DNA     linear        8-JUL-2013
SOURCE      
  ORGANISM  
            This file is created by Biosettia Inc
            http://www.biosettia.com/
FEATURES             Location/Qualifiers
     LTR             7884..8118
                     /label=3'\LTR
     intron          291..1455
                     /label=Intron
     LTR             1..181
                     /label=5'LTR
     misc_feature    7205..7801
                     /label=WPRE
     promoter        1698..3087
                     /label=human\EF1a\promoter
     intron          2126..3068
                     /label=intron
     CDS             6170..7195
                     /label=Hyg
     RBS             5562..6146
                     /label=IRES
     CDS             4799..4852
                     /label=T2A
     CDS             4853..5557
                     /label=RFP\(dTomato)
     CDS             3134..4783
                     /label=Firefly\Luciferase\(FLuc)
BASE COUNT     1918 a      2144 c      2315 g      1741 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 accatggaag atgccaaaaa cattaagaag ggcccagcgc cattctaccc 
     3181 actcgaagac gggaccgccg gcgagcagct gcacaaagcc atgaagcgct acgccctggt 
     3241 gcccggcacc atcgccttta ccgacgcaca tatcgaggtg gacattacct acgccgagta 
     3301 cttcgagatg agcgttcggc tggcagaagc tatgaagcgc tatgggctga atacaaacca 
     3361 tcggatcgtg gtgtgcagcg agaatagctt gcagttcttc atgcccgtgt tgggtgccct 
     3421 gttcatcggt gtggctgtgg ccccagctaa cgacatctac aacgagcgcg agctgctgaa 
     3481 cagcatgggc atcagccagc ccaccgtcgt attcgtgagc aagaaagggc tgcaaaagat 
     3541 cctcaacgtg caaaagaagc taccgatcat acaaaagatc atcatcatgg atagcaagac 
     3601 cgactaccag ggcttccaaa gcatgtacac cttcgtgact tcccatttgc cacccggctt 
     3661 caacgagtac gacttcgtgc ccgagagctt cgaccgggac aaaaccatcg ccctgatcat 
     3721 gaacagtagt ggcagtaccg gattgcccaa gggcgtagcc ctaccgcacc gcaccgcttg 
     3781 tgtccgattc agtcatgccc gcgaccccat cttcggcaac cagatcatcc ccgacaccgc 
     3841 tatcctcagc gtggtgccat ttcaccacgg cttcggcatg ttcaccacgc tgggctactt 
     3901 gatctgcggc tttcgggtcg tgctcatgta ccgcttcgag gaggagctat tcttgcgcag 
     3961 cttgcaagac tataagattc aatctgccct gctggtgccc acactattta gcttcttcgc 
     4021 taagagcact ctcatcgaca agtacgacct aagcaacttg cacgagatcg ccagcggcgg 
     4081 ggcgccgctc agcaaggagg taggtgaggc cgtggccaaa cgcttccacc taccaggcat 
     4141 ccgccagggc tacggcctga cagaaacaac cagcgccatt ctgatcaccc ccgaagggga 
     4201 cgacaagcct ggcgcagtag gcaaggtggt gcccttcttc gaggctaagg tggtggactt 
     4261 ggacaccggt aagacactgg gtgtgaacca gcgcggcgag ctgtgcgtcc gtggccccat 
     4321 gatcatgagc ggctacgtta acaaccccga ggctacaaac gctctcatcg acaaggacgg 
     4381 ctggctgcac agcggcgaca tcgcctactg ggacgaggac gagcacttct tcatcgtgga 
     4441 ccggctgaag agcctgatca aatacaaggg ctaccaggta gccccagccg aactggagag 
     4501 catcctgctg caacacccca acatcttcga cgccggggtc gccggcctgc ccgacgacga 
     4561 tgccggcgag ctgcccgccg cagtcgtcgt gctggaacac ggtaaaacca tgaccgagaa 
     4621 ggagatcgtg gactatgtgg ccagccaggt tacaaccgcc aagaagctgc gcggtggtgt 
     4681 tgtgttcgtg gacgaggtgc ctaaaggact gaccggcaag ttggacgccc gcaagatccg 
     4741 cgagattctc attaaggcca agaagggcgg caagatcgcc gtggctagcg gcgatatcga 
     4801 gggcagaggc agcctgctga catgcggaga tgtggaagag aaccccgggc ccatggtgtc 
     4861 caagggcgag gaagtgatca aagagttcat gcggttcaaa gtgcggatgg aaggcagcat 
     4921 gaacggccac gagttcgaga tcgagggcga gggcgaaggc agaccttacg agggaaccca 
     4981 gaccgccaag ctgaaagtga ccaagggcgg acccctgccc ttcgcctggg atattctgag 
     5041 cccccagttt atgtacggca gcaaagccta cgtgaagcac cccgccgaca tccccgacta 
     5101 caagaagctg agcttccccg agggcttcaa gtgggagaga gtgatgaact tcgaggacgg 
     5161 cggcctcgtg accgtgaccc aggatagctc tctgcaggac ggcaccctga tctacaaagt 
     5221 gaagatgcgg ggcaccaact tcccccccga tggccccgtg atgcagaaaa agaccatggg 
     5281 ctgggaggcc agcaccgaga gactgtaccc tagagatggc gtgctgaagg gcgagatcca 
     5341 ccaggccctg aagctgaagg atggcggcca ctacctggtg gagttcaaga ccatctacat 
     5401 ggccaagaaa cccgtgcagc tgcccggcta ctactacgtg gacaccaagc tggacatcac 
     5461 cagccacaac gaggactaca ccatcgtgga acagtacgag cggagcgagg gccggcacca 
     5521 cctgtttctg tatggcatgg acgagctgta caagtgaact agcccctctc cctccccccc 
     5581 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt 
     5641 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt 
     5701 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa 
     5761 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac 
     5821 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg 
     5881 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt 
     5941 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca 
     6001 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tacacatgct ttacatgtgt 
     6061 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg 
     6121 aaaaacacga tgataatatg gccacactag agatccaccg gtcgccacca tgaaaaagcc 
     6181 tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga 
     6241 cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg 
     6301 tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta 
     6361 tcggcacttt gcatcggccg cgctcccgat tccggaagtg cttgacattg gggaattcag 
     6421 cgagagcctg acctattgca tctcccgccg tgcacagggt gtcacgttgc aagacctgcc 
     6481 tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag gccatggatg cgatcgctgc 
     6541 ggccgatctt agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata 
     6601 cactacatgg cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac 
     6661 tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg 
     6721 ggccgaggac tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt 
     6781 cctgacggac aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga 
     6841 ttcccaatac gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca 
     6901 gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca ggatcgccgc ggctccgggc 
     6961 gtatatgctc cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga 
     7021 tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt 
     7081 cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt 
     7141 actcgccgat agtggaaacc gacgccccag cactcgtccg agggcaaagg aataggcggc 
     7201 cgcgtcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 
     7261 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 
     7321 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 
     7381 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 
     7441 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 
     7501 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 
     7561 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 
     7621 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 
     7681 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 
     7741 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 
     7801 ggaattcgag ctcggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc 
     7861 actttttaaa agaaaagggg ggactggaag ggctaattca ctcccaacga agacaagatc 
     7921 tgctttttgc ttgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg 
     7981 gctaactagg gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag 
     8041 tgtgtgcccg tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag 
     8101 tgtggaaaat ctctagca 
//
