LOCUS Exported 7368 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Insert is a fragment of the Smad4 3'-UTR that contains miR-17 and miR-19 binding sites. To be used for 3'UTR assay.. ACCESSION . VERSION . KEYWORDS pmirGLO-Smad4-1719 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7368) AUTHORS Murphy BL, Obad S, Bihannic L, Ayrault O, Zindy F, Kauppinen S, Roussel MF TITLE Silencing of the miR-17~92 cluster family inhibits medulloblastoma progression. JOURNAL Cancer Res. 2013 Oct 21. PUBMED 24145352 REFERENCE 2 (bases 1 to 7368) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7368 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(60..78) /label=T7 Term /note="T7 terminator, reverse primer" polyA_signal 115..236 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(126..145) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 180..199 /label=SV40pA-R /note="SV40 polyA, reverse primer" promoter 481..838 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(504..524) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 689..824 /label=SV40 ori /note="SV40 origin of replication" primer_bind 751..770 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" regulatory 883..892 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 889..1821 /codon_start=1 /product="Renilla luciferase" /label=hRluc /note="human codon-optimized" /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ" CDS 1873..2664 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="IEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPV LFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSS HLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQG LAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIAL ATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(1924..1943) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2534..2553 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 2728..2776 /note="synthetic polyadenylation signal" promoter 2932..3036 /gene="bla" /label=AmpR promoter CDS 3037..3897 /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 complement(3255..3274) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin complement(4055..4643) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4135..4154) /label=pBR322ori-F /note="pBR322 origin, forward primer" misc_feature 4759..5042 /label=cer region /note="ColE1-derived recombination site that helps to maintain plasmids as monomers" promoter 5099..5598 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(5134..5157) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 5531..5550 /label=mPGK-F /note="Mouse PGK promoter, forward primer" regulatory 5639..5648 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5645..7297 /codon_start=1 /product="firefly luciferase" /label=luciferase /note="synthetic luc2 version of the luciferase gene" /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV" ORIGIN 1 catgcaagct gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac 61 cgctgagcaa taactagcat aaccccttgg ggcggccgct tcgagcagac atgataagat 121 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg 181 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca 241 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gtttttttaa 301 gcaagtaaaa cctctacaaa tgtggtaaaa tcgaatttta acaaaatatt aacgcttaca 361 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgcg 421 gatctgcgca gcaccatggc ctgaaataac ctctgaaaga ggaacttggt taggtacctt 481 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg 541 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg 601 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc 661 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca 721 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc 781 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa 841 gcttgattct tctgacacaa cagtctcgaa ccaaaggctg gagccaccat ggcttccaag 901 gtgtacgacc ccgagcaacg caaacgcatg atcactgggc ctcagtggtg ggctcgctgc 961 aagcaaatga acgtgctgga ctccttcatc aactactatg attccgagaa gcacgccgag 1021 aacgccgtga tttttctgca tggtaacgct gcctccagct acctgtggag gcacgtcgtg 1081 cctcacatcg agcccgtggc tagatgcatc atccctgatc tgatcggaat gggtaagtcc 1141 ggcaagagcg ggaatggctc atatcgcctc ctggatcact acaagtacct caccgcttgg 1201 ttcgagctgc tgaaccttcc aaagaaaatc atctttgtgg gccacgactg gggggcttgt 1261 ctggcctttc actactccta cgagcaccaa gacaagatca aggccatcgt ccatgctgag 1321 agtgtcgtgg acgtgatcga gtcctgggac gagtggcctg acatcgagga ggatatcgcc 1381 ctgatcaaga gcgaagaggg cgagaaaatg gtgcttgaga ataacttctt cgtcgagacc 1441 atgctcccaa gcaagatcat gcggaaactg gagcctgagg agttcgctgc ctacctggag 1501 ccattcaagg agaagggcga ggttagacgg cctaccctct cctggcctcg cgagatccct 1561 ctcgttaagg gaggcaagcc cgacgtcgtc cagattgtcc gcaactacaa cgcctacctt 1621 cgggccagcg acgatctgcc taagatgttc atcgagtccg accctgggtt cttttccaac 1681 gctattgtcg agggagctaa gaagttccct aacaccgagt tcgtgaaggt gaagggcctc 1741 cacttcagcc aggaggacgc tccagatgaa atgggtaagt acatcaagag cttcgtggag 1801 cgcgtgctga agaacgagca gaccggtggt gggagcggag gtggcggatc aggtggcgga 1861 ggctccggag ggattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag 1921 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc 1981 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg 2041 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc 2101 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg 2161 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct 2221 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg 2281 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat 2341 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc 2401 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg 2461 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc 2521 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct 2581 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat 2641 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga 2701 cgcccaacct gccatcacga tggccgcaat aaaatatctt tattttcatt acatctgtgt 2761 gttggttttt tgtgtgaatc gatagcgata aggatcctct ttgcgcttgc gttttccctt 2821 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct 2881 ttctacgtaa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc 2941 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc 3001 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 3061 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 3121 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 3181 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 3241 actttcaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 3301 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 3361 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 3421 gataacactg cggccaactt acttctgaca actatcggag gaccgaagga gctaaccgct 3481 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 3541 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 3601 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 3661 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 3721 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 3781 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 3841 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaattc 3901 gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt 3961 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 4021 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 4081 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 4141 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 4201 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 4261 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 4321 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 4381 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 4441 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag 4501 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 4561 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 4621 cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc 4681 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 4741 cctagatcct tttatagtcc ggaaatacag gaacgcacgc tggatggccc ttcgctggga 4801 tggtgaaacc atgaaaaatg gcagcttcag tggattaagt gggggtaatg tggcctgtac 4861 cctctggttg cataggtatt catacggtta aaatttatca ggcgcgattg cggcagtttt 4921 tcgggtggtt tgttgccatt tttacctgtc tgctgccgtg atcgcgctga acgcgtttta 4981 gcggtgcgta caattaaggg attatggtaa atccacttac tgtctgccct cgtagccatc 5041 gagataaacc gcagtactcc ggccacgatg cgtccggcgt agaggatcga gatctaccgg 5101 gtaggggagg cgcttttccc aaggcagtct ggagcatgcg ctttagcagc cccgctgggc 5161 acttggcgct acacaagtgg cctctggcct cgcacacatt ccacatccac cggtaggcgc 5221 caaccggctc cgttctttgg tggccccttc gcgccacctt ctactcctcc cctagtcagg 5281 aagttccccc ccgccccgca gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt 5341 ctcactagtc tcgtgcagat ggacagcacc gctgagcaat ggaagcgggt aggcctttgg 5401 ggcagcggcc aatagcagct ttgctccttc gctttctggg ctcagaggct gggaaggggt 5461 gggtccgggg gcgggctcag gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc 5521 ggaggcccgg cattctgcac gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc 5581 tcatctccgg gcctttcgac ctgcagccca agcttggcaa tccggtactg ttggtaaagc 5641 caccatggaa gatgccaaaa acattaagaa gggcccagcg ccattctacc cactcgaaga 5701 cgggaccgcc ggcgagcagc tgcacaaagc catgaagcgc tacgccctgg tgcccggcac 5761 catcgccttt accgacgcac atatcgaggt ggacattacc tacgccgagt acttcgagat 5821 gagcgttcgg ctggcagaag ctatgaagcg ctatgggctg aatacaaacc atcggatcgt 5881 ggtgtgcagc gagaatagct tgcagttctt catgcccgtg ttgggtgccc tgttcatcgg 5941 tgtggctgtg gccccagcta acgacatcta caacgagcgc gagctgctga acagcatggg 6001 catcagccag cccaccgtcg tattcgtgag caagaaaggg ctgcaaaaga tcctcaacgt 6061 gcaaaagaag ctaccgatca tacaaaagat catcatcatg gatagcaaga ccgactacca 6121 gggcttccaa agcatgtaca ccttcgtgac ttcccatttg ccacccggct tcaacgagta 6181 cgacttcgtg cccgagagct tcgaccggga caaaaccatc gccctgatca tgaacagtag 6241 tggcagtacc ggattgccca agggcgtagc cctaccgcac cgcaccgctt gtgtccgatt 6301 cagtcatgcc cgcgacccca tcttcggcaa ccagatcatc cccgacaccg ctatcctcag 6361 cgtggtgcca tttcaccacg gcttcggcat gttcaccacg ctgggctact tgatctgcgg 6421 ctttcgggtc gtgctcatgt accgcttcga ggaggagcta ttcttgcgca gcttgcaaga 6481 ctataagatt caatctgccc tgctggtgcc cacactattt agcttcttcg ctaagagcac 6541 tctcatcgac aagtacgacc taagcaactt gcacgagatc gccagcggcg gggcgccgct 6601 cagcaaggag gtaggtgagg ccgtggccaa acgcttccac ctaccaggca tccgccaggg 6661 ctacggcctg acagaaacaa ccagcgccat tctgatcacc cccgaagggg acgacaagcc 6721 tggcgcagta ggcaaggtgg tgcccttctt cgaggctaag gtggtggact tggacaccgg 6781 taagacactg ggtgtgaacc agcgcggcga gctgtgcgtc cgtggcccca tgatcatgag 6841 cggctacgtt aacaaccccg aggctacaaa cgctctcatc gacaaggacg gctggctgca 6901 cagcggcgac atcgcctact gggacgagga cgagcacttc ttcatcgtgg accggctgaa 6961 gagcctgatc aaatacaagg gctaccaggt agccccagcc gaactggaga gcatcctgct 7021 gcaacacccc aacatcttcg acgccggggt cgccggcctg cccgacgacg atgccggcga 7081 gctgcccgcc gcagtcgtcg tgctggaaca cggtaaaacc atgaccgaga aggagatcgt 7141 ggactatgtg gccagccagg ttacaaccgc caagaagctg cgcggtggtg ttgtgttcgt 7201 ggacgaggtg cctaaaggac tgaccggcaa gttggacgcc cgcaagatcc gcgagattct 7261 cattaaggcc aagaagggcg gcaagatcgc cgtgtaattc tagttgttta aactagcggc 7321 cgctagtttt ttttcttttg cacttttgat ctagagtcga cctgcagg //