LOCUS       40924_20071        6486 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Reporter vector pFila, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6486)
  AUTHORS   Bi Y, Zheng X, Shao C, Pan W, Jiang L, Ouyang H.
  TITLE     Construction and application of a built-in dual luciferase reporter 
            for microRNA functional analysis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6486)
  AUTHORS   Bi Y, Zheng X, Shao C, Pan W, Jiang L, Ouyang H.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-OCT-2010) College of Life Science, Wuhan University, 
            Bayi Road, Wuhan, Hubei 430072, P.R. China
REFERENCE   3  (bases 1 to 6486)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6486)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-OCT-2010) College of Life Science, Wuhan University, Bayi Road, 
            Wuhan, Hubei 430072, P.R. China"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        8..759
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     intron          829..961
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1006..1024
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1034..1966
                     /label=Rluc
                     /note="luciferase from the anthozoan coelenterate Renilla 
                     reniformis (sea pansy)"
     misc_feature    1971..1976
                     /label=XbaI restriction site
                     /note="XbaI restriction site"
     misc_recomb     1995..2000
                     /label=ApaI restriction site
                     /note="ApaI restriction site"
     polyA_signal    2028..2149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      2039..2240
                     /label=SV40 late polyadenylation signal (upstream)
                     /note="SV40 late polyadenylation signal (upstream)"
                     /regulatory_class="polyA_signal_sequence"
     promoter        2279..2475
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     CDS             2511..4160
                     /label=luciferase
                     /note="firefly luciferase"
     polyA_signal    complement(4204..4325)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        4695..4799
                     /label=AmpR promoter
     CDS             4800..5657
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      5831..6419
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agatctaaat gagtcttcgg acctcgcggg ggccgcttaa gcggtggtta gggtttgtct
       61 gacgcggggg gagggggaag gaacgaaaca ctctcattcg gaggcggctc ggggtttggt
      121 cttggtggcc acgggcacgc agaagagcgc cgcgatcctc ttaagcaccc ccccgccctc
      181 cgtggaggcg ggggtttggt cggcgggtgg taactggcgg gccgctgact cgggcgggtc
      241 gcgcgcccca gagtgtgacc ttttcggtct gctcgcagac ccccgggcgg cgccgccgcg
      301 gcggcgacgg gctcgctggg tcctaggctc catggggacc gtatacgtgg acaggctctg
      361 gagcatccgc acgactgcgg tgatattacc ggagaccttc tgcgggacga gccgggtcac
      421 gcggctgacg cggagcgtcc gttgggcgac aaacaccagg acggggcaca ggtacactat
      481 cttgtcaccc ggaggcgcga gggactgcag gagcttcagg gagtggcgca gctgcttcat
      541 ccccgtggcc cgttgctcgc gtttgctggc ggtgtccccg gaagaaatat atttgcatgt
      601 ctttagttct atgatgacac aaaccccgcc cagcgtcttg tcattggcga attcgaacac
      661 gcagatgcag tcggggcggc gcggtcccag gtccacttcg catattaagg tgacgcgtgt
      721 ggcctcgaac accgagcgac cctgcagcga cccgcttaaa agcttgattc ttctgacaca
      781 acagtctcga acttaagctg cagaagttgg tcgtgaggca ctgggcaggt aagtatcaag
      841 gttacaagac aggtttaagg agaccaatag aaactgggct tgtcgagaca gagaagactc
      901 ttgcgtttct gataggcacc tattggtctt actgacatcc actttgcctt tctctccaca
      961 ggtgtccact cccagttcaa ttacagctct taaggctaga gtacttaata cgactcacta
     1021 taggctagcc accatgactt cgaaagttta tgatccagaa caaaggaaac ggatgataac
     1081 tggtccgcag tggtgggcca gatgtaaaca aatgaatgtt cttgattcat ttattaatta
     1141 ttatgattca gaaaaacatg cagaaaatgc tgttattttt ttacatggta acgcggcctc
     1201 ttcttattta tggcgacatg ttgtgccaca tattgagcca gtagcgcggt gtattatacc
     1261 agaccttatt ggtatgggca aatcaggcaa atctggtaat ggttcttata ggttacttga
     1321 tcattacaaa tatcttactg catggtttga acttcttaat ttaccaaaga agatcatttt
     1381 tgtcggccat gattggggtg cttgtttggc atttcattat agctatgagc atcaagataa
     1441 gatcaaagca atagttcacg ctgaaagtgt agtagatgtg attgaatcat gggatgaatg
     1501 gcctgatatt gaagaagata ttgcgttgat caaatctgaa gaaggagaaa aaatggtttt
     1561 ggagaataac ttcttcgtgg aaaccatgtt gccatcaaaa atcatgagaa agttagaacc
     1621 agaagaattt gcagcatatc ttgaaccatt caaagagaaa ggtgaagttc gtcgtccaac
     1681 attatcatgg cctcgtgaaa tcccgttagt aaaaggtggt aaacctgacg ttgtacaaat
     1741 tgttaggaat tataatgctt atctacgtgc aagtgatgat ttaccaaaaa tgtttattga
     1801 atcggaccca ggattctttt ccaatgctat tgttgaaggt gccaagaagt ttcctaatac
     1861 tgaatttgtc aaagtaaaag gtcttcattt ttcgcaagaa gatgcacctg atgaaatggg
     1921 aaaatatatc aaatcgttcg ttgagcgagt tctcaaaaat gaacaataat tctagattcc
     1981 gagatatcgg taatgggccc tagagcggcc gcttcgagca gacatgataa gatacattga
     2041 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     2101 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     2161 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aaagcaagta
     2221 aaacctctac aaatgtggta aaatcgataa ggatccaggt ggcacttttc gtgcgatctg
     2281 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     2341 ccgcccagtt ccgcccattc tccgccccat cgctgactaa ttttttttat ttatgcagag
     2401 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     2461 ctaggctttt gcaaaaagct tggcattccg gtactgttgg taaagccacc atggaagacg
     2521 ccaaaaacat aaagaaaggc ccggcgccat tctatccgct ggaagatgga accgctggag
     2581 agcaactgca taaggctatg aagagatacg ccctggttcc tggaacaatt gcttttacag
     2641 atgcacatat cgaggtggac atcacttacg ctgagtactt cgaaatgtcc gttcggttgg
     2701 cagaagctat gaaacgatat gggctgaata caaatcacag aatcgtcgta tgcagtgaaa
     2761 actctcttca attctttatg ccggtgttgg gcgcgttatt tatcggagtt gcagttgcgc
     2821 ccgcgaacga catttataat gaacgtgaat tgctcaacag tatgggcatt tcgcagccta
     2881 ccgtggtgtt cgtttccaaa aaggggttgc aaaaaatttt gaacgtgcaa aaaaagctcc
     2941 caatcatcca aaaaattatt atcatggatt ctaaaacgga ttaccaggga tttcagtcga
     3001 tgtacacgtt cgtcacatct catctacctc ccggttttaa tgaatacgat tttgtgccag
     3061 agtccttcga tagggacaag acaattgcac tgatcatgaa ctcctctgga tctactggtc
     3121 tgcctaaagg tgtcgctctg cctcatagaa ctgcctgcgt gagattctcg catgccagag
     3181 atcctatttt tggcaatcaa atcattccgg atactgcgat tttaagtgtt gttccattcc
     3241 atcacggttt tggaatgttt actacactcg gatatttgat atgtggattt cgagtcgtct
     3301 taatgtatag atttgaagaa gagctgtttc tgaggagcct tcaggattac aagattcaaa
     3361 gtgcgctgct ggtgccaacc ctattctcct tcttcgccaa aagcactctg attgacaaat
     3421 acgatttatc taatttacac gaaattgctt ctggtggcgc tcccctctct aaggaagtcg
     3481 gggaagcggt tgccaagagg ttccatctgc caggtatcag gcaaggatat gggctcactg
     3541 agactacatc agctattctg attacacccg agggggatga taaaccgggc gcggtcggta
     3601 aagttgttcc attttttgaa gcgaaggttg tggatctgga taccgggaaa acgctgggcg
     3661 ttaatcaaag aggcgaactg tgtgtgagag gtcctatgat tatgtccggt tatgtaaaca
     3721 atccggaagc gaccaacgcc ttgattgaca aggatggatg gctacattct ggagacatag
     3781 cttactggga cgaagacgaa cacttcttca tcgttgaccg cctgaagtct ctgattaagt
     3841 acaaaggcta tcaggtggct cccgctgaat tggaatccat cttgctccaa caccccaaca
     3901 tcttcgacgc aggtgtcgca ggtcttcccg acgatgacgc cggtgaactt cccgccgccg
     3961 ttgttgtttt ggagcacgga aagacgatga cggaaaaaga gatcgtggat tacgtcgcca
     4021 gtcaagtaac aaccgcgaaa aagttgcgcg gaggagttgt gtttgtggac gaagtaccga
     4081 aaggtcttac cggaaaactc gacgcaagaa aaatcagaga gatcctcata aaggccaaga
     4141 agggcggaaa gatcgccgtg taattcttga gtcggggcgg ccggccgctt cgagcagaca
     4201 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     4261 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     4321 aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag gtgtgggagg
     4381 ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat ccgtcgaccg
     4441 atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg catgactatc
     4501 gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt gccggcagcg
     4561 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     4621 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4681 gaacatgtga ggtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4741 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     4801 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     4861 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     4921 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     4981 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5041 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5101 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5161 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5221 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5281 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5341 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5401 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     5461 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     5521 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     5581 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     5641 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     5701 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     5761 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5821 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5881 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5941 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     6001 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     6061 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     6121 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     6181 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     6241 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     6301 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     6361 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     6421 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
     6481 ctcgac
//
