LOCUS       Exported                7444 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Luciferase reporter of human Hdm4 3'UTR F2 fragment mutant #5.
ACCESSION   .
VERSION     .
KEYWORDS    Luc-Hdm4-3'UTR-F2d5
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7444)
  AUTHORS   Okada N, Lin CP, Ribeiro MC, Biton A, Lai G, He X, Bu P, Vogel H, 
            Jablons DM, Keller AC, Wilkinson JE, He B, Speed TP, He L
  TITLE     A positive feedback between p53 and miR-34 miRNAs mediates tumor 
            suppression.
  JOURNAL   Genes Dev. 2014 Mar 1;28(5):438-50. doi: 10.1101/gad.233585.113. 
            Epub 2014 Feb 14.
  PUBMED    24532687
REFERENCE   2  (bases 1 to 7444)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7444
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        43..794
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      824..833
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             830..2482
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(896..914)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    2490..2514
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4341..4365)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    4407..4528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4418..4437)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4472..4491
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4776..4793)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4947..5535)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5027..5046)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5706..6566)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6329..6348
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6567..6671)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6698..7153
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6785..6804)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6995..7016
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    7284..7332
                     /note="synthetic polyadenylation signal"
     misc_feature    7346..7437
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     7386..7405
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
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 ttttttagac ggagtctctc tctgtcgccc
     4321 cggcaagggt gggcgcgccg acccagcttt cttgtacaaa gtggtgatct agagtcgggg
     4381 cggccggccg cttcgagcag acatgataag atacattgat gagtttggac aaaccacaac
     4441 tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt
     4501 aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt ttatgtttca
     4561 ggttcagggg gaggtgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa
     4621 aatcgataag gatccgtcga ccgatgccct tgagagcctt caacccagtc agctccttcc
     4681 ggtgggcgcg gggcatgact atcgtcgccg cacttatgac tgtcttcttt atcatgcaac
     4741 tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4801 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4861 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4921 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4981 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5041 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5101 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5161 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5221 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5281 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5341 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5401 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5461 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5521 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5581 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5641 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5701 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5761 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5821 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5881 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5941 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6001 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6061 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6121 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6181 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6241 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6301 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6361 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6421 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6481 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6541 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6601 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6661 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta
     6721 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     6781 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     6841 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     6901 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     6961 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     7021 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     7081 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     7141 acgcttacaa tttgccattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
     7201 cgggcctctt cgctattacg ccagcccaag ctaccatgat aagtaagtaa tattaaggta
     7261 cgggaggtac ttggagcggc cgcaataaaa tatctttatt ttcattacat ctgtgtgttg
     7321 gttttttgtg tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa
     7381 acaaactagc aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctatc
     7441 gata
//
