LOCUS       Exported                7447 bp ds-DNA    circular SYN 21-1月-2014
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    34-163 pGL3-HDM4-3'UTR-F2-mut5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7447)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported 2014年1月21日 from SnapGene Viewer 2.2.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7447
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        43..794
                     /note="HSV TK promoter"
                     /note="herpes simplex virus thymidine kinase promoter"
     CDS             830..2482
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /EC_number="
                     "
                     /note="luciferase"
                     /note="enhanced luc+ version of the luciferase gene"
                     /protein_id="
                     "
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     protein_bind    2490..2514
                     /gene="mutant version of attB"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     3'UTR           2535..4329
                     /note="HDM4-3'UTR-F2-mut5"
     3'UTR           3049..3070
                     /note="miR-34 target site #3"
     3'UTR           3083..3104
                     /note="miR-34 target site #4"
     3'UTR           3900..3919
                     /note="miR-34 target site #5 with mutation"
     protein_bind    complement(4346..4370)
                     /gene="mutant version of attB"
                     /bound_moiety="BP Clonase(TM)"
                     /note="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4410..4531
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(4950..5538)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5709..6569)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6570..6674)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      6701..7156
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    7287..7335
                     /note="synthetic polyadenylation signal"
     misc_feature    7349..7440
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc taaatgagtc ttcggacctc
       61 gcgggggccg cttaagcggt ggttagggtt tgtctgacgc ggggggaggg ggaaggaacg
      121 aaacactctc attcggaggc ggctcggggt ttggtcttgg tggccacggg cacgcagaag
      181 agcgccgcga tcctcttaag cacccccccg ccctccgtgg aggcgggggt ttggtcggcg
      241 ggtggtaact ggcgggccgc tgactcgggc gggtcgcgcg ccccagagtg tgaccttttc
      301 ggtctgctcg cagacccccg ggcggcgccg ccgcggcggc gacgggctcg ctgggtccta
      361 ggctccatgg ggaccgtata cgtggacagg ctctggagca tccgcacgac tgcggtgata
      421 ttaccggaga ccttctgcgg gacgagccgg gtcacgcggc tgacgcggag cgtccgttgg
      481 gcgacaaaca ccaggacggg gcacaggtac actatcttgt cacccggagg cgcgagggac
      541 tgcaggagct tcagggagtg gcgcagctgc ttcatccccg tggcccgttg ctcgcgtttg
      601 ctggcggtgt ccccggaaga aatatatttg catgtcttta gttctatgat gacacaaacc
      661 ccgcccagcg tcttgtcatt ggcgaattcg aacacgcaga tgcagtcggg gcggcgcggt
      721 cccaggtcca cttcgcatat taaggtgacg cgtgtggcct cgaacaccga gcgaccctgc
      781 agcgacccgc ttaaaagctt ggcattccgg tactgttggt aaagccacca tggaagacgc
      841 caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga
      901 gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga
      961 tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc
     1021 agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa
     1081 ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc
     1141 cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac
     1201 cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc
     1261 aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat
     1321 gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga
     1381 gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct
     1441 gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga
     1501 tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
     1561 tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt
     1621 aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag
     1681 tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata
     1741 cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg
     1801 ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga
     1861 gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa
     1921 agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt
     1981 taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa
     2041 tccggaagcg accaacgcct tgattgacaa ggatggatgg ctacattctg gagacatagc
     2101 ttactgggac gaagacgaac acttcttcat cgttgaccgc ctgaagtctc tgattaagta
     2161 caaaggctat caggtggctc ccgctgaatt ggaatccatc ttgctccaac accccaacat
     2221 cttcgacgca ggtgtcgcag gtcttcccga cgatgacgcc ggtgaacttc ccgccgccgt
     2281 tgttgttttg gagcacggaa agacgatgac ggaaaaagag atcgtggatt acgtcgccag
     2341 tcaagtaaca accgcgaaaa agttgcgcgg aggagttgtg tttgtggacg aagtaccgaa
     2401 aggtcttacc ggaaaactcg acgcaagaaa aatcagagag atcctcataa aggccaagaa
     2461 gggcggaaag atcgccgtgt aattctatca caagtttgta caaaaaagca ggctccgcgg
     2521 ccgccccctt caccccacct ggtttctttc tgcatgtccc ctttcacttt caaacctctt
     2581 catttggatg ttaaattata tggtcaccta gttataggta agccttgttc gagttgatat
     2641 cttgattgtg aggaaggatc tgtgtcattg gagcttgttt ctgctgcaac gtgctgtaga
     2701 ctatgaataa tgaaatcaca ccacattacc atcagatttc ttgttttagt tgtcaaatta
     2761 atatttatga ttgttatctt gggcgaaaag ttcagagcag agatgacaaa tcattagaac
     2821 aacgatgaat ttcagtatta cggctaaaaa gttcttctgt ctgaatatta actcactctc
     2881 cttccagtgt acttcacagt aattggtatg cttttttatt taatgcttaa atcaaacttt
     2941 ataaaaatct tagaccagat ctttaatatg gtatgccatt tccccagtct accaatggaa
     3001 tagtatgggt ttctaatcct aggcttgtac aatggattgg agttgagcca tgccagcctc
     3061 cacactgcca ctaacttctg taatgtaaga ttgagtcact gccaagcatt tgaaatatgc
     3121 agttgtgttt taattataat ttatgtatag ttagatgtat gtagtgcatt gtgtggtatt
     3181 atttggtttg taagaattta tttttaaggg tcaaggtcat ttgtaacatt ttgtgtgtgt
     3241 caattcaatg caatgttggc tgccttttga agtctttgat atattggtga atattcttct
     3301 gatctataat acaaagctat gtaatgttac ctcttgactc gcttttgaaa ggaagacaat
     3361 tgttaactag atatttgagt tttttcccct cagaattatg tgaatttctg atatatggct
     3421 ttagatactg tgaatctgtt ttccatttag tcagttatct gcttaaattg ttcagaacta
     3481 tatcctaacg agcaattagt tctgatggtt ctcccagtca tgagtgtgca tgtgtgcaag
     3541 catgttttga tcctgatgct acctttgcta aaaatggcca tagattagga actagctatg
     3601 tttttagaat caaagatgaa ccggtaagct gtctcatgta ccaaacgtga aatttacagt
     3661 gtttacaaat gtctggaatt ttgcactgcc atagggaatg ttaaggttac ttggctggaa
     3721 tttatcagac ttgtgagtaa acaagttgaa gtttagcaga tgagggggaa tattgaggcc
     3781 cctaaggcta aacaaaataa tcagtatctg agatagtggc taatgtggct ccccaggcct
     3841 aatttgggaa cagtttttcc tgattgcttt gagaagtact ttcttttgac agaaattttc
     3901 attctgcttg ctgccatggt attctccctt tataggagcc attggatttc tttccttttg
     3961 tgggaaatgt cccattagca ttttcagatc ttttgatgtg cactaatgcc attattggta
     4021 atgccgttat tggtgaatac agcatagtta aataaactgt tacagtaaat ctacacttgg
     4081 atttgctgca cctctaccaa tagccttttg aatgactgaa agtgttaaca gagaaagagg
     4141 catgtctgca gaaagagata gctaatattt tttggtactt tatctgaaat ccaagatgct
     4201 gcttcccctg caggttgttt tccttcttac gatcctcatt gaatcccctc tgggagcaca
     4261 ggacagttag tagaactctc catttctttt tttttttttt tagacggagt ctctctctgt
     4321 cgccccggca agggtgggcg cgccgaccca gctttcttgt acaaagtggt gatagagtcg
     4381 gggcggccgg ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac
     4441 aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt
     4501 tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt
     4561 tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg
     4621 taaaatcgat aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct
     4681 tccggtgggc gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc
     4741 aactcgtagg acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc
     4801 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4861 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4921 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4981 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5041 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5101 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5161 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5221 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5281 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5341 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5401 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5461 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5521 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5581 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5641 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5701 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5761 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5821 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5881 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5941 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6001 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     6061 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6121 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6181 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6241 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6301 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6361 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6421 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6481 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6541 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6601 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6661 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     6721 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     6781 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6841 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6901 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6961 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     7021 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     7081 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     7141 ttaacgctta caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     7201 gtgcgggcct cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag
     7261 gtacgggagg tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg
     7321 ttggtttttt gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac
     7381 aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct
     7441 atcgata
//
